Peptide sequences and peptide compositions

FIELD: medicine.

SUBSTANCE: group of invention refers to a polypeptide composition containing one or more polypeptides which are immunogenic in a vertebrate so that they induce production in the vertebrate of cells of the immune system able to recognize at least one epitope in a protein fraction of arthropod's saliva with the protein fraction of arthropod's saliva having weight of 20 kDa or less, while the polypeptides are independently specified in a polypeptide sequence SEQ ID NO:1-44 or having 7 amino acids or more partial sequences, methods for making it, using for preparing a drug.

EFFECT: group of invention is effective in treating or preventing a disease propagated by arthropods.

44 cl, 10 dwg, 21 tbl, 4 ex

 

The present invention relates to sequences of the peptides, compositions containing the sequence of the peptides, in particular vaccines distributed arthropod diseases that includes the specified sequence and composition, and applications of sequences. The present invention has, in particular, with respect to vaccines that protect against any one or more distributed mosquito-borne diseases, including, one or more malaria strains.

Protection from disease is important for the survival of all animals, and used for this purpose, a protective mechanism is the immune system of the animal. Therefore, the understanding of the immune system is the key to understanding the development of new or more modern treatments equally for people and animals.

The functioning of the immune system was studied for many years. The system consists of a number of cell types and a variety of molecules, which makes it extremely difficult. Even after many years of research the entire immune system components and their interaction with each other is not fully understood.

Many years ago he admitted that the subject recovered from a specific disease, may acquire in the future some immunity to the disease, not the disease, which is a subject not yet ill. While this fundamental the orbital aspect of the immune system interpreted, considering that the immune system becomes something like "memory" in relation to certain pathogens after exposure to the impact of these pathogens, and the memory is specific for a particular disease.

Little by little it became known that exposure effects are less dangerous variants of a pathogen can induce protection against a more dangerous variants (for example, exposure to the effects of vaccinia virus to protect against variola virus or exposure to the effects of inactivated anthrax bacilli to protect from the living anthrax bacilli). So the idea of vaccination against the disease.

It is now known that the immune system has at least two parts: innate immunity and acquired immunity. Innate immunity is fully functional before the penetration of the pathogen into the system, while acquired immunity is activated upon penetration of the pathogen into the system. She then reveals specific to pathogen attack. The innate immune system includes several components, including phagocytes, such as macrophages, which (in accordance with title) "devour" or absorb a foreign body, such as pathogens.

Usually, but not exclusively, the present invention relates to acquired immune system, and, unless otherwise provided for in case the AEB, "the immune system" in the context of the present invention relates to acquired immune system.

For a more complete understanding of how the functioning of the immune system, should be subjected to careful consideration of the role of its individual components. In respect of the acquired immune system is well known that immunity to pathogens is provided by the action of lymphocytes, which are the most common type of cell in the immune system. There are two types of lymphocytes: b-lymphocytes and T-lymphocytes. They are usually called b-cells and T-cells respectively.

B-cells have the ability to transform into plasma cells that produce antibodies. Antibodies are very important components of the immune system of animals. They are produced in response to some distinctive part of the invasive pathogen (antigen of a pathogen - antigens define here as any alien substance recognized by the immune system) and are usually specific to that pathogen. However, if two of the pathogen are very similar or at least contain the same antigen, antibodies produced against one pathogen may, however, be effective against another pathogen (they can give a "cross-reaction"). This explains why vaccination is iusom ospowiki can protect against variola virus. It is important to understand that the antibodies recognize only a small part of the antigenic molecules of the pathogen, not the whole pathogen. These parts are called epitopes.

T-cells do not have the antibodies, or do not produce them. Instead, they recognize fragments (i.e. epitopes) alien antibody, forming a complex with major histocompatibility complex (MCH) (or in the case of humans the major histocompatibility complex of man (HLA)), through a specialized receptor, known as TCR (receptor T cells). The T-cells are divided into subpopulations, which have or regulatory function, or effector function. Cell effectors associated with the implementation of the" removal of foreign substances. For example, cytotoxic T cells (CTL) cells are the effectors that are able to destroy infected cells, as well as other unwanted species such as tumor cells. On the other hand, regulatory T-cells play a role in helping effector T - and b-cells become more efficient. Due to this feature, these regulatory T cells are often called T-cells"helpers". I think that other regulatory T-cells called T-cells"suppressors"suppress immune responses, but they are less well studied. Regulatory T cells may also interact with components of the VRO is given to the immune system to enhance their activity.

The normal, healthy individual lymphocytes in the immune system remains in an inactive state of "rest" before calling the immune response. If necessary, the immune response of lymphocytes become activated, undergo proliferation and begin to perform their assigned functions. For example, any that rests T-cell, representing on its surface TCR that recognizes an epitope of an invasive pathogen that forms a complex with a MHC molecule, activated, undergo proliferation (this is called "breeding clone"), and the resulting offspring begins to actively perform their assigned effector functions required for the control of invasive organisms.

After completion of the immune response (i.e. removal of pathogens and/or infected cells) lymphocytes return to the quiescent state. However, this state of rest is not equivalent to the original inactive state of rest. Activated, but resting lymphocytes can be quickly subjected to the recruitment and stimulation for proliferation in response to infection by the same or closely related pathogen later.

This ability is activated resting lymphocytes to create a faster and stronger response after the second collision with invasive pathogen effectively provides a "memory" of the immune system. E is splatzie memory of the immune system is the basis for all long-term immunoprophylaxis of medicines (for example, vaccines) and still develop many long-term immunoprophylaxis medicines.

To complete the cells of their functions within the complex systems of the animal on the surface of cells must be "receptors". These receptors are able to "recognize" specific substances that control various important processes, such as activation, proliferation and adhesion to other cells or substances. For example, in the case of immune system receptors on T - and b-cells allow them not only to recognize the antigen, but also to interact with each other and, therefore, to regulate their activity. Without these receptors, the cells would lose a very important means of communication and would not be capable of effective concerted action, which is very important for the immune system of a multicellular organism.

In order for the immune system was capable of specific recognition of a wide range of pathogens present in the environment, and the beginning of the action with respect to such series, there were created two types of highly variable receptor antigens on lymphocytes: antibodies in b-cell receptors of T-cells or TCR in T cells.

To facilitate recognition of a wide range of invasive agents of the immune system in the body there is a huge number what about the different receptors of possible antigens. In fact, a individual there are approximately 1012different b-cells and receptors of T cells. Each b-cell has only one type of receptor, and means for initiating action against a specific pathogen should be selected In the cell having the "best matches" the receptor for the antigen of the pathogen. This process is called "sampling clone". Theoretically can answer only a single clone (monoclonal)or more clones (oligoclonal response), or the many clones (polyclonal response) depending on the number of antigens/epitopes exposed by the disease, and specificity of different selected b-cells against these antigens/epitopes.

Between the types of antigens that can be recognized by b-cells and T-cells, there is a major difference. As far as we know, only the receptors on the surface of b-lymphocytes (i.e. antibodies) capable of direct detection of antigens, such as proteins, viruses and bacteria or foreign molecules dissolved in body fluids. Antibodies can also be produced in a soluble form In cells when they are activated and converted into plasma cells. Antibodies are also called immunoglobulins (abbreviated as Ig). On the other hand, receptors of T-cells only recognize short peptides, also known by the ies as T-cell epitopes, on the surface of cells. These T-cell epitopes are generated in the degradation of large proteins, which are either internal (i.e. occurring in nature proteins of an organism), or not internal (i.e. originating from alien organisms infecting organism). Only epitopes derived from foreign proteins, i.e. antigens, usually able to call the immune response in the body. Once created, these epitopes are associated with a special type of molecule, MHC (major histocompatibility complex), and the resulting complex then prezentuetsya on the cell surface to bind to the receptor of T cells.

It should be clear that due to the destructive nature of the immune response it should be directed only against foreign pathogens, but not against its own cells or proteins of an organism. Thus, the immune system must distinguish between "internal" and "not internal" proteins or cells. An assumption was made that, although effective against internal proteins or cells clones of lymphocytes are produced, they are deleted before it can occur any response. This process is called "deletion clone". It was also assumed that any force against the internal proteins or cells lymphocytes can be saved, but only in "off the hinnon" state. This mechanism is called "clonal tolerance". Whatever was not considered in the process, it remains unclear the exact underlying mechanism that allows lymphoid tissues such as the thymus gland, to identify individual clones of T-cells, acting against the internal proteins or cells from a pool of T-lymphocytes, acting not only against internal proteins or cells.

For many years it was known that the major histocompatibility complex (MHC) plays a key role in the immune system of animals. The MHC molecules contribute to the recognition of antigens by T-cells, as discussed above. There are three main types of MHC molecules, class I, class II and class III. Molecules MHC class I and class II are glycoproteins that are present on the cell surface, whereas molecules MHC class III are usually soluble molecules present inside the cell. There are a large number of different types of MHC molecules. For example, people (whose MHC is called HLA, the major histocompatibility complex of man) there are several hundreds of different alleles of genes encoding MHC molecules, which means that in the human population there are many different types of HLA. MTL different types is usually assigned a different legend, MTL mouse called H-2, rat RT1, and the AOC is the IR RLA. Different region of genes encoding different MHC molecules in an individual, usually called individually, such as HLA-A, HLA-C, etc. in humans.

The MHC molecule is an extremely important molecule of the immune system, because it is the molecule that carries out the presentation of the epitopes of antigens to the immune system. For example, if the T-cell must respond to a specific pathogen, the pathogen must have at least one antigen (e.g. a protein)that has at least one epitope (e.g., peptide protein site), which can communicate with the MHC molecule on the cell surface and, therefore, interact with T-cell that binds to the complex of MHC-peptide. Therefore, the immune response depends on the ability of MHC contact epitope. In the absence of the epitope, which will be contacted by the MNF, or in the absence of T-cells with which to communicate complex MHC-peptide immune response will not arise.

As for internal proteins, one of several epitopes, however, may be capable of binding to the MHC molecule and, consequently, to the possible call of the immune response. In these cases, there shall be provided a specific "signal" for deletions or "off" clones of lymphocytes, acting against the internal proteins.

Despite the races is the current amount of knowledge about the immune system of vertebrates vaccines against many diseases remain elusive. Some pathogens (such as HIV and influenza virus) are fast matirovanie, so epitopes that may be suitable targets for developing a vaccine against one strain, unsuitable after the occurrence of mutations, because the epitope in the new strain has changed. Have been carefully researched other pathogens, such as Plasmodium (the pathogen responsible for malaria), but the target pathogen, which can be used for vaccine development, it has proved difficult to identify, or simply did not provide effective vaccine in vivo.

Pathogens, advances in research are especially necessary are pathogens that spread arthropods and cause so-called "distributed arthropod disease". Such diseases are the leading cause of death worldwide and include, among others, malaria and dengue fever are some of the most significant tools of destruction populations today, particularly in the poorer areas of the world. Examples include, but without limitation, diseases, listed below in table 1.

Table 1
Distributed arthropod diseases, their pathogens and vectors
DiseasePA is Agen (parasite/virus) Which arthropods transmit infections
MalariaPlasmodium sp.Anopheles sp.
FilariasisWuchereria sp.
Brugia sp.
Loa sp.
Mansonella sp.
Anopheles sp.
Aedes sp.
Culex sp.
Mansonia sp.
Chrysops sp.
OnchocerciasisOnchocerca sp.Simulium sp.
Chagas disease
African trypanosomiasis
Trypanosoma sp.Triatoma sp.
Rhodnius sp.
Panstrongylus sp.
Glossina sp.
Tabanus sp.

LeishmaniasisLeishmania sp.Phlebotomus sp.
Lutzomyia sp.
TularemiaFranciscella tulariensisTabanus sp.
Epidemic relapsing fever
Lyme disease
Borrelia sp.Pediculus sp
Ornithodoros sp.
Ixodes sp.
Epidemi the definition typhus Rickettsia sp.Pediculus sp.
Pulex sp.
PlagueYersinia sp.Pulex sp.
Dengue feverDEN-1 - -4 flavivirusAedes sp.
Yellow feverYellow fever flavivirusAedes sp.
Hemagogous spegazzani.
Rift valley feverRift valley virusAedes sp.
Encephalitis, including encephalitis St. Louis encephalitis West Nile, Eastern equine encephalitis virus, encephalitis, and La Crosse, Eastern equine encephalitisFlavi-, Alpha - and BunyavirusesCulex sp.
Ochlerotatus sp.
Aedes sp.
Coquillettidia sp.
Anopheles sp.

Earlier attempts to provide vaccine distributed arthropod diseases included the identification of existing strains of pathogens, and then getting a vaccine specific for this pathogen. Typically, the vaccine was based on In-cell response (antibodies) (and occasionally on the T-cell response), while antimalarial with surface antigens specific strain of the pathogen, against which it was developed. Generally, including surface antigens proteins vary from one caller disease strain to the subsequent strain and is completely different for different pathogens. The result of this is that conventional vaccines usually protect only against a specific pathogen or strain of the pathogen (if at all effective) and will not protect against other strains or new strain, which is a consequence of the mutation. Therefore, a special vaccine required to protect against every disease and often from different and/or new strains that cause the same disease.

For some period of time it is known that immunization against arthropod saliva or against antigens expressed in the gut of arthropods, can protect the individual from infection. The article where this is discussed, include the following highlighted article.

R.G. Titus et al. “The immunomodulatory factors of arthropod saliva and the potential for these factors to serve as vaccine targets to prevent pathogen transmission”, Parasite Immunology, 2006, 28, 131-141. The article provides an overview of the known immunomodulators in the salivary glands of arthropods. Immunomodulatory factors may enhance the transmission of the pathogen arthropods. The saliva of the vector contains a large number of substances activity, which include the ability to inhibition of hemostasis, the narrowing of the blood of Socodevi development of inflammation and immune response. The mosquitoes there are several inhibitors of T cells. If the arthropod saliva enhances infection by pathogens, arthropods which transmit, it should be possible to control transmission of the pathogen through vaccination of the host against molecules (molecules) in the saliva that enhance infection, blocking, thereby amplifying the effects of saliva and thus preventing the establishment of infection by the pathogen in the host. Was cloned gene of maxadrine or MAX, which encodes a strong vasodilator in saliva sandy mosquito, and were investigated activity of the protein product of the gene. The effects of MACH sandy mosquito manifested through the primary effects on phagocytes, and this leads to subsequent immunomodulatory/inhibitory effects on T-cell responses. Mice were vaccinated against MACH by injection MACH in complete Freund's adjuvant, and then MAX in incomplete Freund's adjuvant and then a booster injection of soluble MAX before reaching titers of antibodies against the MACH blood between 1/10000 and 1/20000. Vaccinated mice were explicitly protected from infection.

G.A.T. Targett, “Parasites, arthropod vectors, and immune responses”, Parasite Immunology, 2006, 28, 117-119. This article demonstrates that the extraction of blood by mosquitoes, causes an immune reaction to the saliva of the insect. They include hypersensitivity reactions and the formation of antibodies against the mosquito the century The definition of entities of these antibodies may be of epidemiological significance to control populations of vectors and the use of such entities, to assess the effectiveness of intervention strategies. Getting a vaccine that could reduce the suction of blood, development and/or survival which arthropod vectors or ectoparasites, is an attractive, albeit ambitious idea. One important goal is to control arthropods themselves of ectoparasites, while the second goal is a vaccine against arthropods as a means for inflicting damage to the parasites, which claystone distributes or within himself, or when there is a transfer which is the vertebrate host.

MMA Whitten et al., “Mosquito midguts and malaria: cell biology, compartmentalization and immunology”, Parasite Immunology, 2006, 28, 121-130. Midgut of the mosquito is one of the most difficult environments for the survival and development of Plasmodium. During their attempts to overcome the mid-gut epithelium on the way to the salivary glands movable ookinete quickly identified and marked by means of discriminating factors mosquitoes and made a target for the near destruction of the immune responses that provide recruiting causing the death of factors as of the mid-gut and other tissues surrounding the abdominal cavity. Then the Noe interaction of these factors and parasite is highly specific for species and strain, as the timing and route of invasion of the parasite. Midgut forms a physical barrier that separates and protects the fabric gumiela from digestive enzymes and infectious agents. It consists of a single layer of polarized epithelial cells, one of the two poles found various morphological adaptation to increase the surface area involved in the exchange of molecules. Excellent microcarcinoma apical surface exposed to the lumen of the intestine, and its primary role is the secretion of digestive enzymes and absorption of nutrients. Structural changes initiated by sucking blood, include the formation of a dense non-cellular, chitinous insect peritrophic matrix material (RM), which is secreted by the epithelium of the mid-gut and which is subjected to polymerization under the action of the split food in the form of blood. RM surrounds the blood clot and form a barrier to parasites and attempts bacteria pass through the mid-gut epithelium.

P.F. Billingsley et al. “Immune interactions between mosquitoes and their hosts”, Parasite Immunology, 2006, 28, 143-153. Antibodies directed at the middle gut of mosquitoes, also important in the development of vaccines against mosquitoes. The secretion of saliva by the mosquito during the extraction of blood is essential to the successful location of the blood vessels of the host and impact on games eticheskie and immune response of the host. Methods Western blot turns were used to characterize the recognition of antigens saliva antibodies against mosquito host. While sucking blood Komar swallows immune factors of the host either soluble or cell, which remain active in the middle intestine. In contrast to the antigens of saliva, the owner will typically not be exposed to antigens from the internal organs of the mosquito, which suggests the idea of using these "hidden" antigens as targets for developing a vaccine. The best source of hidden antigens is midgut, because after sucking blood it contains food in the form of blood components with its immune effector molecules and cells. The average ulcer and preparations of the whole body Ae. aegypti induced high titers of antibodies in mice, and a correlation was found increased destruction of mosquitoes that fed on their blood with an antibody which binds to the mid-gut microvilli. Education IgG induced in mice which were injected with cDNA library mid-gut An. Gambiae, and high titers of antibodies were obtained after booster immunization with proteins mid-gut. For mosquitoes feeding on the blood of these mice demonstrated a reproducible decrease in survival and fertility, but, interestingly, cellular, and not humoral responses, apparently, were the CTE is responsible for mosquitocide effects. These studies are faced with problems. Even within the experiments, the instability can be high and difficult to achieve reproducibility of the effect. Immunization is a complex mixture or protein extracts of mid-gut, or a cDNA library mid-gut meant that protective, which is the target antigens have not been identified.

However, despite this amount of knowledge is still not developed an effective vaccine against disseminated arthropod diseases, which uses this mechanism of action. A further significant problem associated with existing vaccines distributed arthropod diseases based or b-cell or T-cell responses, is that each of them only protects from one pathogen or the biggest one of the existing strain of the pathogen and does not provide protection against possible future pathogens or from a variety of pathogens. There is a desperate need for a vaccine that protects against multiple distributed arthropod diseases, including large-scale destruction, such as malaria and dengue fever.

Currently, the authors of the present invention were able to identify the sequence of specific immunogenic peptides that are present in proteins of arthropod saliva, which is s can provide protection from all distributed arthropod pathogens, and developed vaccines against diseases that they cause, using the identified sequence. Thus, the authors of the present invention have developed peptides that are applicable in vaccines induce immune response and, in particular, rapid secondary immune response against distributed arthropod diseases.

Accordingly, the present invention is to resolve the problems associated with the prior art presented above. A further purpose of this invention is the provision of polypeptide composition, which is able to induce the vertebrate immune response (e.g., cellular responses, including T-cell response and/or b-cell response) against many distributed arthropod diseases, i.e. diseases caused by many pathogens or strains of pathogens. A further purpose of the present invention is the provision of vaccines distributed arthropod diseases, using the polypeptide compositions of the present invention.

Accordingly, the present invention provides polypeptide composition comprising one or more polypeptides that are immunogenic in the spinal, so they cause spinal products of cells of the immune system (or activate the production of such cells), with osobnych to recognize at least one epitope in the protein fraction of arthropod saliva, moreover, the protein fraction of arthropod saliva has mass, component of 40 kDa or less, and polypeptides independently selected from:

(a) sequences of the polypeptides SEQ ID NO:1-44 or having 7 amino acids or more subsequences of these sequences:

SEQ ID NO:1 HLTLFTVAVLLLAAAALLLLLPPAYSTTLTPP

SEQ ID NO:2 PLSYCHLFLTHTLARALSFSRSDCL

SEQ ID NO:3 KNVFFALLLVVLVCCLVSVQGNEI

SEQ ID NO:4 KLLVLLICLFFYHTHCTTAYLWLAMGV

SEQ ID NO:5 FLKGSFPRFQMCVMLIGFFSSAKCL

SEQ ID NO:6 NDYQALLGLCCPWIDLAAADLPMRRHAKA

SEQ ID NO:7 FYSVGKLVKVLLVMAVCCLLLCTAPTGADPL

SEQ ID NO:8 MKFAFAFVLIALFAVFAVSQALPQPEQAAA

SEQ ID NO:9 DGASAITKIVLELTPEQAAAV

SEQ ID NO:10 TLFIFLVCCQIPLFGIMSSDSADPFYWIRVILA

SEQ ID NO:11 GRVMCLLRLMSTLLVVLSIVGK

SEQ ID NO:12 LYSGYRLLVLLVMTVCCLLLFIAPTGADPLPGQTQRTL

SEQ ID NO:13 MYCVIKGKTGGYCNSEGLCTCRAEDLHFLLKPIINKD

SEQ ID NO:14 NAEDPRTELIGCGSVLFHLAANRLSLQLEEFAVCKR

SEQ ID NO:15 ALIGLLLCSVQSVTANDPVDALGACSGNLFGLLMTRL

SEQ ID NO:16 SKLFVLAFLCLALVVVVQSAPQYARGDVPT

SEQ ID NO:17 SMLVAFATLSVALVVVVAIPANFNYGGGGGYFINGTGQ

SEQ ID NO:18 IYEKLPAYLSEVSARVNVLQVSLQHDLPNLQ

SEQ ID NO:19 EMKLAKVALVTISLWFMAWTPYLVINFTGI

SEQ ID NO:20 LLPAKVIPDKTAAYVAYGGQETLVEHVEVLV

SEQ ID NO:21 FYTCFLGTSSLAGFKNAVDYDELLKAG

SEQ ID NO:22 VLEVLGFVEDNGELVFQELLGVLKMVDPDGD

SEQ ID NO:23 KLTPTVVVVLLCLTFVADALTIQELRAQIAQQRIQQRYGVTVATT

SEQ ID NO:24 SLSDYGLIELKEHCLECCQKDTEADSKLKVYPAAVLEV

SEQ ID NO:25 TYICFILHGVSEIIPQQQKKTMKFLLLVASVLCLVLI

SEQ ID NO:26 RYFVVIALICPLIIVETLAV

SEQ ID NO:27 LLLYLDAADLRRALHQYQLLAAQGDRHLPQQIVKFV

SEQ ID NO:28 VLLTPALQAYIMDEHNLNRSNIALGRIRPYPSAVKMP

SEQ ID NO:29 VLKGETHKALKLKDGGHYLVEFKSIYM

SEQ ID NO:30 VLHSMLVNASLAEMVKESYQTHGADGRMVVRMLKFVRLLP

SEQ ID NO:31 RVRALRALLETLLQHQGEQNNDVYLIRLAHET

SEQ ID NO:32 ELQQALSSLNAGSGSCAEVFNAYLPVHNKYIGVSRKI

SEQ ID NO:33 KFYRLISTLLVVVVIAPRHQCSPFFFQYNRPYL

SEQ ID NO:34 NYVPDVSALEQDIIEVDPETKEMLKHLDFNNIVVQL

SEQ ID NO:35 QYSMECLEAAEPKYLDGLKTLADETAQC

SEQ ID NO:36 EYAQVTKMLGNGRLEAMCFGVKRLCHIRGKL

SEQ ID NO:37 KLFLTLLSTLSVAMVFALPAHHHSRG

SEQ ID NO:38 ELEEARLVAEELEERQQELDYLKRYLVGRLQAV

SEQ ID NO:39 SYFLTVCLLALVQSETVQD

SEQ ID NO:40 AMTNANLVGLTISLAYAIFFLLYTPPTGRSS

SEQ ID NO:41 SFAWLLYGIILRSNFLVVQNLMALALSAVQLSLFII

SEQ ID NO:42 AFPFISGFLSCFMWLKYGVLTEESTLILVNFIGSAL

SEQ ID NO:43 GLLCCCLAVLFFASPLTMLAHVIR

SEQ ID NO:44 LLLAMVLLPLLLLESVVPYAAAEKVW

(b) sequences that are limited to the following amino acid residues of the protein of arthropod saliva or having 7 amino acids or more subsequences of these sequences:

remnants 2-33 >gi|18389913|gb|AAL68793.1|AF457563_1 hypothetical protein 16 [Anopheles gambiae]

residues 2-26 >gi|18389909|gb|AAL68791.1|AF457561_1 hypothetical protein 14 [Anopheles gambiae]

residues 2-25 >gi|18389907|gb|AAL68790.1|AF457560_1 hypothetical protein 13 [Anopheles gambiae]

remnants 10-36 >gi|18389903|gb|AAL68788.1|AF457558_1 hypothetical protein 11 [Anopheles gambiae]

residues 2-26 >gi|62546227|gb|AAX86005.1|hyp3.5-precursor [Anopheles gambiae]

the remnants of the 14-42 gi|18389899|gb|AAL68786.1|AF457556_1 7-like protein in the salivary glands [Anopheles gambiae]

residues 3-33 >gi|18389911|gb|AAL68792.1|AF457562_1 hypothetical protein 15 [Anopheles gambiae]

residues 1-30 >gi|62546225|gb|AAX86004.1|hyp6.3 - precursor [Anopheles gambiae]

remnants 34-54 >gi|62546225|gb|AAX86004.1|hyp6.3 - precursor [Anopheles gambiae]

remnants of 38-70 >gi|17026153|emb|CAD12038.1|Sec61 protein [Anopheles gambiae]

residues 2-23 >gi|62546223|gb|AAX86003.1|hyp6.2 - precursor [Anopheles gambiae]

remnants 17-54 >gi|18389915|gb|AAL68794.1|AF457564_1 hypothetical protein 17 [Anopheles gambiae]

remnants 57-93 >gi|87080391|gb|ABD18596.1|Def is Cena [Anopheles gambiae]

the remnants of 22-57 >gi|18389901|gb|AAL68787.1|AF457557_1 hypothetical protein 10 [Anopheles gambiae]

remnants 7-43 >gi|18389905|gb|AAL68789.1|AF457559_1 hypothetical protein 12 [Anopheles gambiae]

remnants 3-32 >gi|4127344|emb|CAA76832.1|cE5-protein [Anopheles gambiae]

remnants 3-40 >gi|4210617|emb|CAA10259.1|SG2 protein [Anopheles gambiae]

remnants 91-121 >gi|4127309|emb|CAA76820.1|hypothetical protein [Anopheles gambiae]

remnants 65-94 >gi|4375824|emb|CAA76825.1|opsin has been performed [Anopheles gambiae]

remnants 41-71 >gi|62546233|gb|AAX86008.1|unknown protein [Anopheles gambiae]

remnants 117-143 >gi|3378531|emb|CAA03872.1|D7r2 protein [Anopheles gambiae]

remnants 63-93 >gi|3378529|emb|CAA03871.1|D7r3 protein [Anopheles gambiae]

remnants 23-67 >gi|18389893|gb|AAL68783.1|AF457553_1 muzykopodobnyh protein [Anopheles gambiae]

remnants 43-80 >gi|18389881|gb|AAL68777.1|AF457547_1 selenoprotein [Anopheles gambiae]

remnants 6-42 >gi|18389879|gb|AAL68776.1|AF457546_1 protein with Mm 30 kDa [Anopheles gambiae]

remnants 4-23 >gi|18378603|gb|AAL68639.1|AF458073_1 D7-related protein 5 [Anopheles gambiae]

the remnants of 20-55 >gi|18389897|gb|AAL68785.1|AF457555_1 1-like protein 4 salivary glands [Anopheles gambiae]

remnants 59-95 >gi|18389883|gb|AAL68778.1|AF457548_1 antigen 5-related protein 1 [Anopheles gambiae]

remnants 158-184 >gi|83016748|dbj|BAE53441.1|DsRed [synthetic constructs]

remnants 37-76 >gi|18389895|gb|AAL68784.1|AF457554_1 1-like protein 3 salivary glands [Anopheles gambiae]

remnants 191-222 >gi|18389895|gb|AAL68784.1|AF457554_1 1-like protein 3 salivary glands [Anopheles gambiae]

remnants 113-149 >gi|18389891|gb|AAL68782.1|AF457552_1 long form D7-protein [Anopheles gambia]

residues 1-37 >emb|CAC35527.1|gSG9-protein [Anopheles gambiae]

remnants 81-120 >sp|Q9U9L1|RS17_ANOGA protein S17 40S subunit of the ribosome

remnants 111-142 >emb|CAC35523.1|gSG7 protein [Anopheles gambiae]

remnants 32-67 >gb|AAD47075.1|AF164151_1 factor 4C (1A) of translation initiation [Anopheles gambiae]

residues 1-29 >emb|CAC35519.1|gSG2-like protein [Anopheles gambiae]

remnants 106-142 >emb|CAC35451.1|hypothetical protein [Anopheles gambiae]

residues 6-28 >emb|CAC35524.1|D7r4 protein [Anopheles gambiae]

remnants 70-104 >ref|XP_001230998.1|ENSANGP00000014906 [Anopheles gambiae strain PEST]

remnants 174-213 >ref|XP_316361.2|ENSANGP00000012984 [Anopheles gambiae strain PEST]

remnants 41-80 >ref|XP_314140.3|ENSANGP00000015780 [Anopheles gambiae strain PEST]

remnants 126-153 >ref|XP_314140.3|ENSANGP00000015780 [Anopheles gambiae strain PEST]

residues 5-34 >emb|CAC35522.1|gSG6 protein [Anopheles gambiae]

(c) and sequences of polypeptides that are homologous to 85% or more of the one or more sequences of (a) or (b) and contained in one or more of the following databases: GeneBank, the data Bank of protein structure (PDB), SwissProt, protein Information resource (PIR), the Foundation for the study of proteins (PRF) or broadcasts of coding sequences (CDS) of these databases.

CDS is an abbreviation for "coding sequence", i.e. a region of nucleotides that corresponds to the sequence of amino acids in the predicted protein. CDS includes initiating codon and the stop codon, therefore, to yousie sequence begins with "ATG", and ends with a stop codon. Expressively sequence, including 5'-UTR, 3'-UTR, introns, are not expressed due to a shift in the reading frame that is not included in CDS. Please note that the CDS does not match the actual current sequence of mRNA. As a result, the broadcast CDS is a protein that could be obtained if they were translated all the codons between the initiating codon and the stop codon.

PDB means the Bank of data on the structure of the protein. This database (http://www.rcsb.org/pdb/home/home.do) is supported by the Collaboration for the study of structural bioinformatics (RCSB), not getting profit consortium, specializing in improving understanding of the functioning of biological systems through the study of three-dimensional structures of biological macromolecules.

Information protein resource (PIR) (http://pir.georgetown.edu an integral common bioinformatics resource, created in 1984, the National Foundation for biomedical research (NBRF) as a resource that can help researchers to identify and interpret information about protein sequence (C.H. Wu, L.S. Yeh, G. Huang, L. Arminski, Castro-Alvear j, Chen y, Hu z, Kourtesis p, Ledley RS, Suzek B.E., Garrett L., Vinayaka C.R., Zhang J., Backer W.C. (2003) “The Protein Information Resource”, Nucleic Acids Res. 31(1): 345-7).

PRF is the online database that is supported by the Fund for the study of proteins (PRF) (http://www.prf.or.jp/en/index.shtml). Base Yes the data contains information related to amino acids, peptides and proteins collected from scientific journals, data sequences of peptides and proteins, data on synthetic compounds and molecular aspects of protein.

GenBank is a database of gene sequences NIH (http://www.ncbi.nlm.nih.gov/), annotated collection of all publicly available DNA sequences. A new version is created every two months. GenBank is part of the International collaboration on creation of a database concerning a nucleotide sequence, which consists of a data Bank of DNA of Japan (DDBJ), European molecular biology laboratory (EMBL) and GenBank at the National center for biotechnology information.

Swissprot (also called UniProtKB/Swiss-Prot) is a curated database of protein sequences (http://expasy.org/sprot supported by the Institute of bioinformatics, Switzerland (SIB). The database aims to provide high level comments (such as the description of the function of the protein, its domain structure, post-translational modifications, variants, etc. to provide redundancy to a minimum, and integration with other databases at a high level.

These databases are updated weekly or monthly, and the above sequence apply to sequences in the databases at the time under the Chi of this application. When searching in databases of sequences having the desired degree of homology, it is possible to use either method, depending on eligibility criteria. However, preferably can be used the BLASTP program [BLAST and its derivatives which program (e.g., BLASTP) is a free software].

In another embodiment, instead referred to above in paragraph (C) homology of 85% (or more) are also included sequences of the polypeptides in the composition of these databases, which have at least 85% amino acids, which is shared (on the basis of amino acid identity and position in the sequence) with the amino acid part of the sequence in (a) or (b), the length of which is 8 amino acids or more, and preferably 8 amino acids up to one-third the length of the sequence in (a) or (b). In other words, for the sequence in (a) or (b), the length of which is 30 amino acids, is also included sequence database, if it shares 85% or more of its amino acids with any part of the sequence in (a) or (b)having a length of 8 amino acids or more, preferably 8-10 amino acids. Similarly, if the length of the sequence in (a) or (b) is 60 amino acids, also included a sequence database, if the shares on 85% or more of its amino acids with any part of the sequence in (a) or (b), having a length of 8 amino acids or more, preferably 8-20 amino acids. Compliance amino acids is not necessarily consistent. For example, in the case of sequences of length 20 amino acids in (a) or (b)if the corresponding sequence database parts 17 or more amino acids in the relevant provisions, it is included, even if not all of these provisions are consistent.

Typically, the polypeptides in the composition are not full size (or whole) proteins of arthropod saliva. Under the full-size (or whole) means that the polypeptides really contain all of the amino acid residues present in any of the naturally occurring proteins of arthropod saliva.

Thus, the polypeptide is a polypeptide, which may include any of the above sequences as a whole (or may include portions of any of the above sequences of length at least 7 or more residues). The polypeptide must be immunogenic in spinal. Typically, this immunogenicity is such that the polypeptides cause spinal products of cells of the immune system capable of recognizing at least one epitope in the protein fraction of arthropod saliva. Therefore, if the polypeptide induces T-cell or b-cell response, it is suitable for all kinds of what is immunogenic in the spinal. In the alternative case, the polypeptide can be an epitope of the T-lymphocyte helper (Th) or may be an epitope of b-lymphocyte.

One way to determine whether the immunogenicity of the polypeptide, are presented below in experiment 2. However, the present invention is not limited to such methods, and a trained professional can choose any known method of determining immunogenicity, as desired.

Particularly preferably, when the polypeptides polypeptide compositions independently selected from SEQ ID NO:1-6, 20, 28, 30-32 and 35 or having 7 amino acids or more subsequences of these sequences or sequences of polypeptides that are homologous to 85% or more of one of these sequences and are contained in one or more of the following databases: GeneBank, the data Bank of protein structure (PDB), SwissProt, protein Information resource (PIR), the Foundation for the study of proteins (PRF) or broadcasts of coding sequences (CDS) of these databases.

Usually, but not exclusively, the polypeptide composition of the present invention includes 2 or more polypeptides, preferably from 2 to 10 or more polypeptides preferably 2-6 polypeptides. However, the composition may include one polypeptide and additional non-polypeptide components, if desired.

Typically, the polypeptide compositions in accordance with the present invention the protein fraction of arthropod saliva has mass, component 40 kDa or less, 30 kDa or less, or more preferably 20 kDa or less. The fraction can also be mass component of 20-40 kDa, 20-30 kDa or 10-20 kDa.

In another embodiment of the present invention the polypeptide composition of the present invention includes SEQ ID NO:131 or includes one or more subsequences of SEQ ID NO:131, having 7 amino acids or more, or contains sequences of polypeptides that are homologous to 85% or more of one of these sequences and are contained in one or more of the following databases: GeneBank, the data Bank of protein structure (PDB), SwissProt, protein Information resource (PIR), the Foundation for the study of proteins (PRF) or broadcasts of coding sequences (CDS) of these databases.

SEQ ID NO:131

FLKGSFPRFQMCVMLIGFFSSAKCLFYSVGKLVKVLLVMAVCCLLLCTAPTGADPLMKFAFAFVLIALFAVFAVSQALPQPEQAAAGRVMCLLRLMSTLLVVLSIVGKLYSGYRLLVLLVMTVCCLLLFIAPTGADPLPGQTQRTLALIGLLLCSVQSVTANDPVDALGACSGNLFGLLMTRLSKLFVLAFLCLALVVVVQSAPQYARGDVPTLLPAKVIPDKTAAYVAYGGQETLVEHVEVLVRYFVVIALICPLIIVETLAVVLLTPALQAYIMDEHNLNRSNIALGRIRPYPSAVKMPVLHSMLVNASLAEMVKESYQTHGADGRMVVRMLKFVRLLPRVRALRALLETLLQHQGEQNNDVYLIRLAHETELQQALSSLNAGSGSCAEVFNAYLPVHNKYIGVSRKIQYSMECLEAAEPKYLDGLKTLADETAQCSFAWLLYGIILRSNFLVVQNLMALALSAVQLSLFIIAFPFISGFLSCFMWLKYGVLTEESTLILVNFIGSAL

In another embodiment of the present invention the polypeptide composition of the present invention includes one or more sequences selected from SEQ ID nos:1-4, 6, 9, 10, 13, 14, 17-19, 21-25, 27, 29, 33, 34, 36-40, 43 and 44, or includes one or more imeushih amino acids or more amino acid subsequences of these sequences, or contains sequences of polypeptides, which homologous 85% or more of one of these sequences and are contained in one or more of the following databases: GeneBank, the data Bank of protein structure (PDB), SwissProt, protein Information resource (PIR), the Foundation for the study of proteins (PRF) or broadcasts of coding sequences (CDS) of these databases.

In a further embodiment, the polypeptide composition of the present invention includes SEQ ID NO:132 or includes one or more subsequences of SEQ ID NO:132, having 7 amino acids or more, or contains sequences of polypeptides that are homologous to 85% or more of one of these sequences and are contained in one or more of the following databases: GeneBank, the data Bank of protein structure (PDB), SwissProt, protein Information resource (PIR), the Foundation for the study of proteins (PRF) or broadcasts of coding sequences (CDS) of these databases.

SEQ ID NO:132

HLTLFTVAVLLLAAAALLLLLPPAYSTTLTPPPLSYCHLFLTHTLARALSFSRSDCLKNVFFALLLVVLVCCLVSVQGNEIKLLVLLICLFFYHTHCTTAYLWLAMGVNDYQALLGLCCPWIDLAAADLPMRRHAKADGASAITKIVLELTPEQAAAVTLFIFLVCCQIPLFGIMSSDSADPFYWIRVILAMYCVIKGKTGGYCNSEGLCTCRAEDLHFLLKPIINKDNAEDPRTELIGCGSVLFHLAANRLSLQLEEFAVCKRSMLVAFATLSVALVVVVAIPANFNYGGGGGYFINGTGQIYEKLPAYLSEVSARVNVLQVSLQHDLPNLQEMKLAKVALVTISLWFMAWTPYLVINFTGIFYTCFLGTSSLAGFKNAVDYDELLKAGVLEVLGFVEDNGELVFQELLGVLKMVDPDGDKLTPTVVVVLLCLTFVADALTIQELRAQIAQQRIQQRYGVTVATTSLSDYGLIELKEHCLECCQKDTEADSKLKVYPAAVLEVTYICFILHGVSEIIPQQQKKTMKFLLLVASVLCLVLILLLYLDAADLRRALHQYQLLAAQGDRHLPQQIVKFVVLKGETHKALKLKDGGHYLVEFKSIYMKFYRLISTLLVVVVIAPRHQCSPFFFQYNRPYLNYVPDVSALEQDIIEVDPETKEMLKHLDFNNIVVQLEYAQVTKMLGNGRLEAMCFDGVKRLCHIRGKLKLFLTLLSTLSVAMVFALPAHHHSRGELEEALVAEELEERQQELDYLKRYLVGRLQAVSYFLTVCLLALVQSETVQDAMTNANLVGLTISLAYAIFFLLYTPPTGRSSGLLCCCLAVLFFASPLTMLAHVIRLLLAMVLLPLLLLESVVPYAAAEKVW

In a still further embodiment, the polypeptide composition of the present invention includes SEQ ID NO:133 or includes one or more subsequences of SEQ ID NO:133, having 7 amino acids or more, or contains sequences of polypeptides that are homologous to 85% or more of one of these sequences and are contained in one or more of the following databases: GeneBank, the data Bank of protein structure (PDB), SwissProt, protein Information resource (PIR), the Foundation for the study of proteins (PRF) or broadcasts of coding sequences (CDS) of these databases.

SEQ ID NO:133

HLTLFTVAVLLLAAAALLLLLPPAYSTTLTPPPLSYCHLFLTHTLARALSFSRSDCLKNVFFALLLVVLVCCLVSVQGNEIKLLVLLICLFFYHTHCTTAYLWLAMGVFLKGSFPRFQMCVMLIGFFSSAKCLNDYQALLGLCCPWIDLAAADLPMRRHAKA

In a still further embodiment, the polypeptide composition of the present invention includes SEQ ID NO:134 or includes one or more subsequences of SEQ ID NO:134, having 7 amino acids or more, or contains sequences of polypeptides that are homologous to 85% or more of one of these sequences and are contained in one or more of the following databases: GeneBank, the data Bank of protein structure (PDB), SwissProt, protein Information resource (PIR), the Foundation for the study of proteins (PRF) or broadcasts of coding sequences (CDS) of these databases.

SEQ ID NO:134

LLPAKVIPDKTAAYVAYGGQETLVEHVEVLVVLLTPALQAYIMDEHNLNRNIALGRIRPYPSAVKMPVLHSMLVNASLAEMVKESYQTHGADGRMVVRMLKFVRLLPRVRALRALLETLLQHQGEQNNDVYLIRLAHETELQQALSSLNAGSGSCAEVFNAYLPVHNKYIGVSRKIQYSMECLEAAEPKYLDGLKTLADETAQC

It is important that in all variants of implementation of the present invention can be used, or separately, or in combination, any combination of the listed sequences. Particularly preferred polypeptides are polypeptides having the sequence of SEQ ID NO:1-6, and/or a sequence having sequence SEQ ID NO:20, 28, 30-32 and 35. Particularly preferably, when present, any one or more of these sequences, and especially preferably, when there are all of SEQ ID nos:1-6 and/or all of SEQ ID NO:20, 28, 30-32 and 35.

The authors of the present invention found that the above sequence include the epitope or multiple epitopes, which can provide protection against distributed arthropod diseases for a wide range of vertebrate populations. The bites of arthropods cause the host's immune response to components of saliva, which is characterized by a Th2 phenotype (i.e. inhibition of the production of IFN-γ and activation of IL-4) (Mbow et al., 1998). We found that this immune response, together with antihemostatic effect of many of these molecules saliva, facilitates the transfer of parasites and increases in General (Dhar and Kumar, 2003) and, in particular, contributes to infection with Leishmania (Kamhawi et al., 2000). It is established that, on the contrary, the activation of the cellular immune response, characterized by ovalicin the th production of IFN-γ and IL-12, both of which are cytokines of the Th1 type, at the site of infection (i.e. the site of the bite) induces protection against infection with Leishmania major through the bite of infected sand mosquitoes (Kamhawi et al., 2000).

Without limitation to any theory believe that immunization with saliva proteins, leading to activation of the Th1 type response, will lead to rapid detection of antigens in saliva at the site of bite cells of the immune system (such as activated cytotoxic T cells (CTL) and T-cell helper type 1) and production of IFN-γ. This cytokine (1) stimulates greater production of IFN-γ as T-and NK-cells, (2) enhances the bactericidal activity of macrophages, (3) induces isotype switching to IgG2a and increased production of IgG2a In cells, and (4) the production of many cytokines (such as TNF-α, interleukin (IL)-12 and IL-18), which together trigger a cascade of immune reactions that lead to the deaths of intracellular parasites. Below are the links, in which it outlines.

Mbow M.L., Bleyenberg J.A., Hall, L.R. & Titus R.G. 1998. Phlebotomus papatasi sandfly salivary gland lysate down-regulates a Th1, but up-regulates a Th2, response in mice infected with Leishmania major, J. Immunol., 161; 5571-5577.

Dhar R., Kumar N. 2003. Role of mosquito salivary glands. Cur. Sci., 85, 1308-1313.

Kamhawi, S., Y. Belkaid, Modi G., E. Rowton, Sacks D., 2000. Protection against cutaneous leishmaniasis resulting from bites of uninfected sand flies. Science 290, 1351-1354.

L. Malaguarnera, Musumeci S. 2002. The immune response to Plasmodium falciparum malaria, Lancet Infect. Dis. Aug; 2(8): 472-8.

Ka is discussed above, the above sequences were identified after sequence analysis of the saliva of Anopheles gambiae. For a qualified professional will be apparent that the present invention applies not only to these sequences and their epitopes, but also on large sequences in proteins of arthropod saliva containing these sequences, and sequences that are homologous to these sequences, and therefore, have immunogenic activity. Thus, in the scope of the present invention are sequences that are homologous to some degree of consensus sequences. This homology allows the replacement of, for example, up to 3 amino acids in the 8-dimensional epitope (homology 62.5%) or 9-d, 10-d or 11-dimensional epitope. Preferably, when no more than 10 such substitutions are identified in the sequence of the present invention, the corresponding full-sized sequences SEQ ID NO:1-44 (homology 66.6% for 30-measure). Preferably, such substitutions are conservative substitutions in accordance with known schema of replacement.

The present invention also provides a polypeptide containing one or more sequences defined by the following sequences SEQ ID NO:1-44 or restricted by the following amino acid ostad the mi protein saliva arthropods, or containing one or more having 7 amino acids or more subsequences of these sequences, or contains sequences of polypeptides that are homologous to 85% or more of one of these sequences and are contained in one or more of the following databases: GeneBank, the data Bank of protein structure (PDB), SwissProt, protein Information resource (PIR), the Foundation for the study of proteins (PRF) or broadcasts of coding sequences (CDS) of these databases.

SEQ ID NO:1 HLTLFTVAVLLLAAAALLLLLPPAYSTTLTPP

SEQ ID NO:2 PLSYCHLFLTHTLARALSFSRSDCL

SEQ ID NO:3 KNVFFALLLVVLVCCLVSVQGNEI

SEQ ID NO:4 KLLVLLICLFFYHTHCTTAYLWLAMGV

SEQ ID NO:5 FLKGSFPRFQMCVMLIGFFSSAKCL

SEQ ID NO:6 NDYQALLGLCCPWIDLAAADLPMRRHAKA

SEQ ID NO:7 FYSVGKLVKVLLVMAVCCLLLCTAPTGADPL

SEQ ID NO:8 MKFAFAFVLIALFAVFAVSQALPQPEQAAA

SEQ ID NO:9 DGASAITKIVLELTPEQAAAV

SEQ ID NO:10 TLFIFLVCCQIPLFGIMSSDSADPFYWIRVILA

SEQ ID NO:11 GRVMCLLRLMSTLLVVLSIVGK

SEQ ID NO:12 LYSGYRLLVLLVMTVCCLLLFIAPTGADPLPGQTQRTL

SEQ ID NO:13 MYCVIKGKTGGYCNSEGLCTCRAEDLHFLLKPIINKD

SEQ ID NO:14 NAEDPRTELIGCGSVLFHLAANRLSLQLEEFAVCKR

SEQ ID NO:15 ALIGLLLCSVQSVTANDPVDALGACSGNLFGLLMTRL

SEQ ID NO:16 SKLFVLAFLCLALVVVVQSAPQYARGDVPT

SEQ ID NO:17 SMLVAFATLSVALVVVVAIPANFNYGGGGGYFINGTGQ

SEQ ID NO:18 IYEKLPAYLSEVSARVNVLQVSLQHDLPNLQ

SEQ ID NO:19 EMKLAKVALVTISLWFMAWTPYLVINFTGI

SEQ ID NO:20 LLPAKVIPDKTAAYVAYGGQETLVEHVEVLV

SEQ ID NO:21 FYTCFLGTSSLAGFKNAVDYDELLKAG

SEQ ID NO:22 VLEVLGFVEDNGELVFQELLGVLKMVDPDGD

SEQ ID NO:23 KLTPTVVVVLLCLTFVADALTIQELRAQIAQQRIQQRYGVTVATT

SEQ ID NO:24 SLSDYGLIELKEHCLECCQKDTEADSKLKVYPAAVLEV

SEQ ID NO:25 TYICFILHGVSEIIPQQQKKTMKFLLLVASVLCLVLI

SEQ ID NO:26 RYFVVIALICPLIIVETLAV

SEQ ID NO:27 LLLYLDAADLRRALHQYQLLAAQGDRHLPQQIVKFV

SEQ ID NO:28 VLLTPALQAYIMDEHNLNRSNIALGRIRPYPSAVKMP

SEQ ID NO:29 VLKGETKALKLKDGGHYLVEFKSIYM

SEQ ID NO:30 VLHSMLVNASLAEMVKESYQTHGADGRMVVRMLKFVRLLP

SEQ ID NO:31 RVRALRALLETLLQHQGEQNNDVYLIRLAHET

SEQ ID NO:32 ELQQALSSLNAGSGSCAEVFNAYLPVHNKYIGVSRKI

SEQ ID NO:33 KFYRLISTLLVVVVIAPRHQCSPFFFQYNRPYL

SEQ ID NO:34 NYVPDVSALEQDIIEVDPETKEMLKHLDFNNIVVQL

SEQ ID NO:35 QYSMECLEAAEPKYLDGLKTLADETAQC

SEQ ID NO:36 EYAQVTKMLGNGRLEAMCFDGVKRLCHIRGKL

SEQ ID NO:37 KLFLTLLSTLSVAMVFALPAHHHSRG

SEQ ID NO:38 ELEEARLVAEELEERQQELDYLKRYLVGRLQAV

SEQ ID NO:39 SYFLTVCLLALVQSETVQD

SEQ ID NO:40 AMTNANLVGLTISLAYAIFFLLYTPPTGRSS

SEQ ID NO:41 SFAWLLYGIILRSNFLVVQNLMALALSAVQLSLFII

SEQ ID NO:42 AFPFISGFLSCFMWLKYGVLTEESTLILVNFIGSAL

SEQ ID NO:43 GLLCCCLAVLFFASPLTMLAHVIR

SEQ ID NO:44 LLLAMVLLPLLLLESVVPYAAAEKVW

remnants 2-33 >gi|18389913|gb|AAL68793.1|AF457563_1 hypothetical protein 16 [Anopheles gambiae]

residues 2-26 >gi|18389909|gb|AAL68791.1|AF457561_1 hypothetical protein 14 [Anopheles gambiae]

residues 2-25 >gi|18389907|gb|AAL68790.1|AF457560_1 hypothetical protein 13 [Anopheles gambiae]

remnants 10-36 >gi|18389903|gb|AAL68788.1|AF457558_1 hypothetical protein 11 [Anopheles gambiae]

residues 2-26 >gi|62546227|gb|AAX86005.1|hyp3.5 - precursor [Anopheles gambiae]

the remnants of the 14-42 gi|18389899|gb|AAL68786.1|AF457556_1 7-like protein in the salivary glands [Anopheles gambiae]

residues 3-33 >gi|18389911|gb|AAL68792.1|AF457562_1 hypothetical protein 15 [Anopheles gambiae]

residues 1-30 >gi|62546225|gb|AAX86004.1|hyp6.3 - precursor [Anopheles gambiae]

remnants 34-54 >gi|62546225|gb|AAX86004.1|hyp6.3 - precursor [Anopheles gambiae]

remnants of 38-70 >gi|17026153|emb|CAD12038.1|Sec61 protein [Anopheles gambiae]

residues 2-23 >gi|62546223|gb|AAX86003.1|hyp6.2 - precursor [Anopheles gambiae]

remnants 17-54 >gi|18389915|gb|AAL68794.1|AF457564_1 hypothetical protein 17 [Anopheles gambiae]

remnants 57-93 >gi|7080391|gb|ABD18596.1|defensin [Anopheles gambiae]

the remnants of 22-57 >gi|18389901|gb|AAL68787.1|AF457557_1 hypothetical protein 10 [Anopheles gambiae]

remnants 7-43 >gi|18389905|gb|AAL68789.1|AF457559_1 hypothetical protein 12 [Anopheles gambiae]

remnants 3-32 >gi|4127344|emb|CAA76832.1|cE5-protein [Anopheles gambiae]

remnants 3-40 >gi|4210617|emb|CAA10259.1|SG2 protein [Anopheles gambiae]

remnants 91-121 >gi|4127309|emb|CAA76820.1|hypothetical protein [Anopheles gambiae]

remnants 65-94 >gi|4375824|emb|CAA76825.1|opsin has been performed [Anopheles gambiae]

remnants 41-71 >gi|62546233|gb|AAX86008.1|unknown protein [Anopheles gambiae]

remnants 117-143 >gi|3378531|emb|CAA03872.1|D7r2 protein [Anopheles gambiae]

remnants 63-93 >gi|3378529|emb|CAA03871.1|D7r3 protein [Anopheles gambiae]

remnants 23-67 >gi|18389893|gb|AAL68783.1|AF457553_1 muzykopodobnyh protein [Anopheles gambiae]

remnants 43-80 >gi|18389881|gb|AAL68777.1|AF457547_1 selenoprotein [Anopheles gambiae]

remnants 6-42 >gi|18389879|gb|AAL68776.1|AF457546_1 protein with Mm 30 kDa [Anopheles gambiae]

remnants 4-23 >gi|18378603|gb|AAL68639.1|AF458073_1 D7-related protein 5 [Anopheles gambiae]

the remnants of 20-55 >gi|18389897|gb|AAL68785.1|AF457555_1 1-like protein 4 salivary glands [Anopheles gambiae]

remnants 59-95 >gi|18389883|gb|AAL68778.1|AF457548_1 antigen 5-related protein 1 [Anopheles gambiae]

remnants 158-184 >gi|83016748|dbj|BAE53441.1|DsRed [synthetic constructs]

remnants 37-76 >gi|18389895|gb|AAL68784.1|AF457554_1 1-like protein 3 salivary glands [Anopheles gambiae]

remnants 191-222 >gi|18389895|gb|AAL68784.1|AF457554_1 1-like protein 3 salivary glands [Anopheles gambiae]

remnants 113-149 >gi|18389891|gb|AAL68782.1|AF457552_1 long form D7-protein [Anopheles gambia]

remnants 3-35 >gi|13537676|emb|CAC35527.1|gSG9-protein [Anopheles gambiae]

remnants 83-118 >sp|Q9U9L1|RS17_ANOGA protein S17 40S subunit of the ribosome and gb|AAD47077.1|AF164153_1 ribosomal protein S17 [Anopheles gambiae]

remnants 113-140 >emb|CAC35523.1|gSG7 protein [Anopheles gambiae]

remnants 34-65 >gb|AAD47075.1|AF164151_1 factor 4C (1A) of translation initiation [Anopheles gambiae]

remnants 2-27 >emb|CAC35519.1|gSG2-like protein [Anopheles gambiae]

remnants 108-140 >emb|CAC35451.1|hypothetical protein [Anopheles gambiae] >gb|EAU75730.1|ENSANGP00000031975 [Anopheles gambiae strain PEST]

residues 8-26 >emb|CAC35524.1|D7r4 protein [Anopheles gambiae] >gb|AAK84945.1|D7-related protein 4 [Anopheles gambiae]

remnants 72-102 >ref|XP_001230998.1|ENSANGP00000014906 [Anopheles gambiae strain PEST] and gb|EAU76798.1|ENSANGP00000014906 [Anopheles gambiae strain PEST]

remnants 176-211 >ref|XP_316361.2|ENSANGP00000012984 [Anopheles gambiae strain PEST] >gb|EAA10852.2|ENSANGP00000012984 [Anopheles gambiae strain PEST]

remnants 43-78 >ref|XP_314140.3|ENSANGP00000015780 [Anopheles gambiae strain PEST] and gb|EAA09398.3|ENSANGP00000015780 [Anopheles gambiae strain PEST]

remnants 128-151 >ref|XP_314140.3|ENSANGP00000015780 [Anopheles gambiae strain PEST] and gb|EAA09398.3|ENSANGP00000015780 [Anopheles gambiae strain PEST]

remnants 7-32 >emb|CAC35522.1|gSG6 protein [Anopheles gambiae]

Preferably, the polypeptide is not the full-length protein of arthropod saliva.

Referred to in the present invention, the numbering sequence is determined in accordance with generally accepted principles. Therefore, the numbering starts with 1 from the set of codon translation initiation (ATG). He shall correspond to methionine (M), for a segment of the genome that encodes protein of interest. In other words, it starts with 1 in accordance with the methionine, is detected as the first amino acid of interest protein sequence used and defined databases that reflect the sequence (i.e. GenBank, SwissProt etc).

Read more the present invention will be described by way of example only with reference to the following figures, in which:

figures 1 and 9 are shown gels after isoelectric focusing (IEF) of the salivary glands of Anopheles gambiae (with staining of Kumasi blue), obtained in accordance with the protocols presented in the examples;

figure 2 shows electrophoresis in SDS-page IEF-gel 1 (painted silver) - red rectangle shows the location of one of the targeted proteins (<30 kDa);

on figa-3I shows the following data on the effect of the vaccine on fertility mosquitoes:

3A: Percentage of mosquitoes that received food

3B: Average number of produced eggs

3C: Average number eggs

3D: Percentage hatching

3E: the Average number of larvae

3F: the Average number of pupae

3G: Percentage pupation

3H: Percentage departure

3I: Average number of adult mosquitoes

Figure 4 shows data on the survival percentage of mosquitoes after which Armenia blood of immunized mice.

Figure 5 shows the production of IFN-gamma after 96 hours of stimulation in vitro antigens in experiment 3.

Figure 6 illustrates the formation of the Ig as a whole in the sera antigens in accordance with experiment 3.

Figure 7 illustrates the formation of the Ig as a whole in the sera on the AGS mixture at day 21 in accordance with experiment 4.

On Fig demonstrates subjected to immunization AGS-mix group in experiment 4, showing increased survival compared with mosquitoes in the control group subjected to immunization NRP-mixture.

The above-described polypeptides generally include one or more (preferably two or more epitopes. Preferably these epitopes are T-cell epitopes, such as epitopes of cytotoxic T lymphocytes (CTL), but can also contain b-cell epitopes. Typically, the polypeptide is immunogenic relative to the protein of arthropod saliva and preferably relatively many such proteins. In the context of the present invention, it is assumed that the polypeptide, immunogenic relative to protein saliva arthropods, means polypeptide that is part of a protein of arthropod saliva and induces an immune system response. One way of determining whether a polypeptide such immunogenicity below in experiment 2. Takanassee the invention is not limited to such methods, and qualified professional may choose any known method of determining immunogenicity, if desired.

In the present invention the polypeptide composition comprises one or more of the above defined sequences. Generally, two, three, four, five or more of such sequences may be present in the polypeptide composition, if desired. The more of these epitopes is present, the greater the width of the protection afforded in the population are people and/or animals individuals with different HLA or MHC. This is especially true if enabled epitopes derive from the saliva of many different arthropods or separated proteins in saliva of different kinds of arthropods and can, therefore, protect against diseases caused by a variety of arthropods. Typically, the polypeptide composition comprises 10 polypeptides or less, preferably 6 polypeptide or less and usually 2-10 polypeptide, and more preferably 2-6 polypeptides.

Polypeptide composition in accordance with the present invention may also contain one or more additional sequences that are not epitopes, if desirable. As a rule, additional sequences originate from one or more proteins of arthropod saliva, predpochtite is correctly chosen from sequences SEQ ID NO:45-85 or subsequences. These sequences can be between two or more sequences (epitopes), described above, or may be located on one or both ends of the polypeptide. The presence of such additional series shall not affect the operation of the polypeptide, provided that the polypeptide as a whole does not become too large, making noise in the presentation of epitopes in the immune system of the spinal.

In the most preferred embodiments, the implementation of additional sequences of the above proteins are sequences, which are sequences (or inside them):

SEQ ID NO:45 - >gi|18389913|gb|AAL68793.1|AF457563_1 hypothetical protein 16 [Anopheles gambiae]

MHLTLFTVAVLLLAAAALLLLLPPAYSTTLTPPAPPRLSHLGITIGRI

SEQ ID NO:46 - >gi|18389909|gb|AAL68791.1|AF457561_1 hypothetical protein 14 [Anopheles gambiae]

MPLSYCHLFLTHTLARALSFSRSDCLKFSEKRLLFSGSKTFPTTLL

SEQ ID NO:47 - >gi|18389907|gb|AAL68790.1|AF457560_1 hypothetical protein 13 [Anopheles gambiae]

MKNVFFALLLVVLVCCLVSVQGNEIIQNVVKRSIPLRQLILQHNALDDSNSDSGSQ

SEQ ID NO:48 - >gi|18389903|gb|AAL68788.1|AF457558_1 hypothetical protein 11 [Anopheles gambiae]

MCIFFQAGIKLLVLLICLFFYHTHCTTAYLWLAMGVEAKSIKARGTAHSKSRTSTN

SEQ ID NO:49 - >gi|62546227|gb|AAX86005.1|hyp3.5 - precursor [Anopheles gambiae]

MFLKGSFPRFQMCVMLIGFFSSAKCLMCFADWEGMLLMTMEVFDFQLIVFTPVLKRS

SEQ ID NO:50 - >gi|18389899|gb|AAL68786.1|AF457556_1 7-like protein in the salivary glands [Anopheles gambiae]

MAGESQKNARSKQNDYQALLGLCCPWIDLAAADLPMRRHAKAREAINFLLQAHEAGPNEEPSLPA

SEQ ID NO:51 - >gi|18389911|gb|AAL68792.1|AF457562_1 gipoteticheski protein 15 [Anopheles gambiae]

MKFYSVGKLVKVLLVMAVCCLLLCTAPTGADPLPGRDRNTIANKSKDKKASAPKHSLGTGARMALTGGGVLGGVLTNM

SEQ ID NO:52 - >gi|62546225|gb|AAX86004.1|hyp6.3 - precursor [Anopheles gambiae]

MKFAFAFVLIALFAVFAVSQALPQPEQAAASSNDGASAITKIVLELTPEQAAAVQKMGGRGFWPIMMKSVKKIMAIGCDLIDC

SEQ ID NO:53 - >gi|17026153|emb|CAD12038.1|Sec61 protein [Anopheles gambiae]

MGIKFLEIIKPFCGILPEIAKPERKIQFREKVLWTAITLFIFLVCCQIPLFGIMSSDSADPFYWIRVILASNRGTLM

SEQ ID NO:54 - >gi|62546223|gb|AAX86003.1|hyp6.2 - precursor [Anopheles gambiae]

MGRVMCLLRLMSTLLVVLSIVGKKTNAAPQVTEAPGNVGSTYSPMADIGRLATGATKLFGQFWNTGTRFGTELSRRTFDFLRVKK

SEQ ID NO:55 - >gi|18389915|gb|AAL68794.1|AF457564_1 hypothetical protein 17 [Anopheles gambiae]

MAGDIQLFSTRETTMKLYSGYRLLVLLVMTVCCLLLFIAPTGADPLPGQTQRTLGYRGNDKRATPPMHSLGSGARMAMTGGGILGGIFSAL

SEQ ID NO:56 - >gi|87080391|gb|ABD18596.1|defensin [Anopheles gambiae]

MDQCSVPRLCIIIMKSFIAAAVIALICAIAVSGTTVTLQSTCKLFTADVVSSITCKMYCVIKGKTGGYCNSEGLCTCRAEDLHFLLKPIINKD

SEQ ID NO:57 - >gi|18389901|gb|AAL68787.1|AF457557_1 hypothetical protein 10 [Anopheles gambiae]

MRFLSVLTVGLLVWVGVFATVNAEDPRTELIGCGSVLFHLAANRLSLQLEEFAVCKRSNPGYDCSDSIHRAISDLQQGLFDLNHCTKDIR

SEQ ID NO:58 - >gi|18389905|gb|AAL68789.1|AF457559_1 hypothetical protein 12 [Anopheles gambiae]

MRFCCVALIGLLLCSVQSVTANDPVDALGACSGNLFGLLMTRLQQMVEDFTACRQEATANDPQHDRSDSIQRAKVDLQQQLVNYSYCTKNIQ

SEQ ID NO:59 - >gi|4127344|emb|CAA76832.1|cE5-protein [Anopheles gambiae]

MASKLFVLAFLCLALVVVVQSAPQYARGDVPTYDEEDFDEESLKPHSSSPSDDGEEEFDPSLLEEHADAPTARDPGRNPEFLRNSNTDEQASAPAASSSDS

SEQ ID NO:60 - >gi|4210617|emb|CAA10259.1|SG2 protein [Anopheles gambiae]

MKSMLVAFATLSVALVVVVAIPANFNYGGGGGYFINGTGQSFNFSGESNGTSIPGLPDFGSFLPNLGNLTQQFGGSSGAFPQFSIPSWTNFTDAFTSILPFFGNGQGGGFPFFG

SEQ ID NO:61 - >gi|4127309|emb|CAA76820.1|hypothetical protein [Anopheles gambiae]

MTPLIATLAACALTLSIVHSRGLPESSDKLEACGQHYGXLLKASTTWNEKECNGSTKLAACVVSEHEQAYRELKQRCQEAHDERTAKVNAIYEKLPAYLSEVSARVNVLQVSLQHDLPNLQE

SEQ ID NO:62 - >gi|4375824|emb|CAA76825.1|opsin [Anopheles gambiae]

PDVAEPLVHHHLRHLRVLAAAADHHLLVHLHPEGCVRSREEHARAGQEGNVASLRTQEAQNTSTEMKLAKVALVTISLWFMAWTPYLVINFTGIFKAAPISPLATIRGSLFAKANVYNPIVYG

SEQ ID NO:63 - >gi|62546233|gb|AAX86008.1|unknown protein [Anopheles gambiae]

MATTWIPTSVHGPYPPHMVPGGVDSDGAQIFVGRAHHAGDLLPAKVIPDKTAAYVAYGGQETLVEHVEVLVHKQLIWDTASAGQVPLGAVVGGHTSDGEILYVGRAYHEGSQTIGKVQCSHNCIYIPYGGAEVSVPTYEVLCER

SEQ ID NO:64 - >gi|3378531|emb|CAA03872.1|D7r2 protein [Anopheles gambiae]

MFKKLLLSVGLVWCLISLGQARKESTVEECEKNIGDSLKDRVCELRQYTPVSSDDMDKHMQCVLEVVGFVDGNGEVKESVLLELLQRVDSGVNHAANMKKCVTEASTSGSDKKANTFYTCFLGTSSLAGFKNAVDYDELLKAGKMQTSDP

SEQ ID NO:65 - >gi|3378529|emb|CAA03871.1|D7r3 protein [Anopheles gambiae]

MFGKLLPCAILLWCLFSLGQARQEETVEECERNIPASLKERVCELRQYTPVQGKDMDSHMQCVLEVLGFVEDNGELVFQELLGVLKMVDPDGDHAGSMKKCNGEAEKVDTSSKANTFYTCFLGTSSAQAFKYAVDYVXAXRAGKLDMGTTFNAGQV

SEQ ID NO:66 - >gi|18389893|gb|AAL68783.1|AF457553_1 mezinarodni protein [Anopheles gambiae]

AGGFSLFEALKQTTTRGEMFRRKLTPTVVVVLLCLTFVADALTIQELRAQIAQQRIQQRYGVTVATTSAATTTAATTSAATTSEATTTAAASTTQASDSDNTTTTAEATTTTEAQTTSSSDNSTTTEAAATTTAASETTADSSSTGTTSVEAGLRAQYRDQVRQQAIERALARAAAFG

SEQ ID NO:67 - >gi|18389881|gb|AAL68777.1|AF457547_1 selenoprotein [Anopheles gambiae]

MRLFAITCLLFSIVTVIGAEFSAEDCRELGLIKSQLFCSACSSLSDYGLIELKEHCLECCQKDTEADSKLKVYPAAVLEVCTCKFGAYPQIQAFIKSDRPAKFPNLTIKYVRGLDPIVKLMDEQGTVKETLSINKWNTDTVQEFFETRLAKVEDDDYIKTNRV

SEQ ID NO:68 - >gi|18389879|gb|AAL68776.1|AF457546_1 protein with Mm 30 kDa [Anopheles gambiae]

MAGAITYICFILHGVSEIIPQQQKKTMKFLLLVASVLCLVLIVSARPADDTSDQESSTELSDDAGAEEGAEDAGSDAEADAGAADGEEGATDTESGAEGDDSEMDSAMKEGEEGAGSDDAVSGADDETEESKDDAEEDSEEGGEEGGDSASGGEGGEKESPRNTYRQVHKLLKKIMKVDTKD

SEQ ID NO:69 - >gi|18378603|gb|AAL68639.1|AF458073_1 D7-related protein 5 [Anopheles gambiae]

MEWRYFVVIALICPLIIVETLAVSDCVRHVSESARNTVCDVRQYRVTKGVEADRYVQCFMTALGFADESGSIQRSNVLTALDAVETHDGVYTDAVDVCLSKAKKLPGTERSGYFFSCMLRTESALNFRDAVELQELRVASKWPEGERFDRSKVQQMMRELNSQLRC

SEQ ID NO:70 - >gi|18389897|gb|AAL68785.1|AF457555_1 1-like protein 4 salivary glands [Anopheles gambiae]

GREAIETMRTEQRNHRQQLLLLYLDAADLRRALHQYQLLAAQGDRHLPQQIVKFVYAAPRHENRRLENLLDLVRQLPARQDQRTLYQLLQPEIMKRPAQNQSTLAMLTALEMGQVVEGNGELKKQQDAMYQLVLKRWMFLCLAGQYREIVQFATKHPRLFE

SEQ ID NO:71 - >gi|18389883|gb|AAL68778.1|AF457548_1 antigen 5-related protein 1 [Anopheles gmbiae]

MAIWIVCATLLLAVLSVVSVGGQYCSSDLCPRGGPHVGCNPPSSSGGPTCQGKQKARKVLLTPALQAYIMDEHNLNRSNIALGRIRPYPSAVKMPTLTWDPELASLADANARSCNYGHDRCRATKKFPYAGQNIAITQFFGYRFTEKDLIHKFVSSWWSEYLDARPEHVRKYPSSYSG

SEQ ID NO:72 - >gi|83016748|dbj|BAE53441.1|DsRed [synthetic construct]

MKLASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVKLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHHLFLRSRAPPPPPLT

SEQ ID NO:73 - >gi|18389895|gb|AAL68784.1|AF457554_1 1-like protein 3 salivary glands [Anopheles gambiae]

MAGQRHLIEQAWQYGAQLQHELMLTSMESDRVQRALVLHSMLVNASLAEMVKESYQTHGADGRMVVRMLKFVRLLPGADERVAVYKQLAELLKSNGQDGRFPAVIFSTDVRQLEDRYKPDHAQYEGKVVERWLAELQAGTFHEVVEFARDYPEYFARVEEPLYETLKQQWSAEGLDRMVSFPNALPVGVQRVRALRALLETLLQHQGEQNNDVYLIRLAHETGRVEATVGQADAAVRQALDDVKKLFEQFKYQRGFPDYEALYKLFKGL

SEQ ID NO:74 - >gi|18389891|gb|AAL68782.1|AF457552_1 long form D7-protein [Anopheles gambiae]

MIVPRVLLFILLELFVQATQAFKALDPEEAWYVYERCHEDHLPSGPNRETYLKTWKFWKLEPNDAVTHCYVKCTLAGLQMYDEKTNTFKPETVPVQHEAYKSFTEVESSKVNELQQALSSLNAGSGSCAEVFNAYLPVHNKYIGVSRKIYHGTVDSVAKIYEAKPEIKKQEESFFAYCAKKALGANGKEGYKKIRDYELADSAEFRNAMDCVFRGFRYMDDSGLKVDEVVRDFNLINKSDLEPEVRSVLASCTGTHAYDYYSCLLNSSVKEDFRNAFYFHELRSANYGYLAMGKVYEGPEKVKEELKKLNY

SEQ ID NO:75 - >emb|CAC35527.1|gSG9-protein [Anopheles gambiae]

MCKFYRLISTLLVVVVIAPRHQCSPFFFQYNRPYLSQPSSQLASTAANVVQRSNVTVALGNRINTDTALDDYGTRV

SEQ ID NO:76 - >sp|Q9U9L1|RS17_ANOGA protein S17 40S subunit of the ribosome

MGRVRTKTIKKASKVIIEKYYTRLTMDFDTNKRIVEEVAIIPTKPLRNKIAGFVTHLMKRLRHSQVRGISIKLQEEERERRDNYVPDVSALEQDIIEVDPETKEMLKHLDFNNIVVQLTNPTAPGYSNRRN

SEQ ID NO:77 - >emb|CAC35523.1|gSG7 protein [Anopheles gambiae]

MHAKPAFVLIALGVICLLQTTPTSASTNHVQQLMKVFRSMTQNFDYTKKPSYLQRAKYGVQNQLRNPLVQKAGNLPKSAKLSDGCLKQMVARVTDLEASFYASFSYNCHDHDQYSMECLEAAEPKYLDGLKTLADETAQCMRDQQ

SEQ ID NO:78 - >gb|AAD47075.1|AF164151_1 factor 4C (1A) of translation initiation [Anopheles gambiae]

MPKNKGKGGKNRRRGKNENESEKRELIFKEDEQEYAQVTKMLGNGRLEAMCFDGVKRLCHIRGKLRKKVWINQGDIILIGLRDYQDSKADVILKYTPDEARNLKTYGEFPESVRINETVTFVENDMDDDIEFGDDYSSSEEGDAIDAI

SEQ ID NO:79 - >emb|CAC35519.1|gSG2-podobn the th protein [Anopheles gambiae]

MKLFLTLLSTLSVAMVFALPAHHHSRGGDGSSANSTGNSDNNSAGVPDFGFNSQSNVPGFGNGQQPGQQQQGQQGQGFPFFGQGQSGFPSFGNRLQPFFGQNQQGQDGDAQQGRGVPFFGQGGGQGGIPSFGSGQQNGGVPFLGNGQGQSGFPSFGNGQQGGNFPFFG

SEQ ID NO:80 - >emb|CAC35451.1|hypothetical protein [Anopheles gambiae]

MKLYAFALVLCVGLAVGAEVDSVPEVPSDLQQQLDELQLADKPEAPVDDAEQPLPPNGDELPEDAPEPVPEDGSPDEEHLEEEQEEEAEADEEEADESESEESEESDELEEARLVAEELEERQQELDYLKRYLVGRLQAVAILDRRVRPAVIRRPWIRRPWIRRPG

SEQ ID NO:81 - >emb|CAC35524.1|D7r4 protein [Anopheles gambiae]

MIRQVIISYFLTVCLLALVQSETVQDCENKLPPSLKSRLCEIRRYEIIEGPEMDKHIHCVMRALDFVYEDGRGDYHKLYDPLNIIELDKRHDVNLEKCIGECVQVPTSERAHVFYKCLLKSTTGRTFKKVFDLMELKKAGKVPQHQRYTAEFVQIMKDYDKALNC

SEQ ID NO:82 - >ref|XP_001230998.1|ENSANGP00000014906 [Anopheles gambiae strain PEST]

MEAISEALQPYKEQVGMAAGILTVGQMFSGCFVCNDIRKKGTTDGFSAMPFVGGCGLTVLFLQHGMLMNDSAMTNANLVGLTISLAYAIFFLLYTPPTGRSSYWRQVGGTALFTITLLGYVKVENPSVVEDRFGMIITVLMLALIGQPLFGLPDIIRRKSTEGLPFAMILSGTIVGLSWLLYGVILNNVFVVCQNLAAVTLSGIQLALFAIYPSKAAPPSKKRE

SEQ ID NO:83 - >ref|XP_316361.2|ENSANGP00000012984 [Anopheles gambiae strain PEST]

MESIAVALQPYKDTVGLTAAIVTVVQFFSGVLALNAIRRQGNTRGFSALPFLGGTVFCLLNIQFGQMLRDDGMIRVNFIGLALNLLYVCGFYLYTEGPAKTAVWGQIGLAGALTAGVLSYVQYEDPQLVEFRFGLILTGLLWTLVGMPLLGLGDILKKKSTEGLPFPIIFLGAVVSFAWLLYGIILRSNFLVVQNLMALALSAVQLSLFIIFPSGAAKPPPTPAKKRN

SEQ ID NO:84 - >ref|XP_314140.3|ENSANGP00000015780 [Anopheles gambiae strain PEST]

MDGIMSKGSLASLATVATVLQFLTGTVICNRYIRKKSTGDTSAFPFISGFLSCFMWLKYGVLTEESTLILVNFIGSALFFSYTVVFFIFCVNKREVIRQMMVISCIILSATLYTLFETDDEKSIRVIGLLCCCLAVLFFASPLTMLAHVIRTQNTDSLPFPIIMASFFVCLLWTAYGVLIGDRFIQIPNLLGGILAGIQLTLYVIYPKKKASFSGGPRYSPLVSENPIL

SEQ ID NO:85 - >emb|CAC35522.1|gSG6 protein [Anopheles gambiae]

MAIRVELLLAMVLLPLLLLESVVPYAAAEKVWVDRDKVYCGHLDCTRVATFKGERFCTLCDTRHFCECKETREPLPYMYACPGTEPCQSSDRLGSCSKSMHDVLCDRIDQAFLEQ

The peptides of the present invention, such as the peptides with SEQ ID NO:1-44 and peptides within SEQ ID NO:131-134 described above preferably include one or more additional amino acids at one or both ends to facilitate their processing in vaccines. Typically, these on the additional amino acids are amino acids, adjacent to each of the ends of the SEQ ID NO:1-44, as demonstrated above in large proteins SEQ ID NO:45-86 (these large proteins contain sequences SEQ ID NO:1-44). Preferably the number of additional amino acids at each end is 1-5, more preferably 1-3 and most preferably 2 at each end. In each of these cases, if at the end of the sequences SEQ ID NO:1-44 is less than two additional amino acids, additional amino acids include no more than the remaining amino acids (amino acids) on this end. Particularly preferred sequences of this type, corresponding to SEQ ID NO:1-44, are the following sequence:

SEQ ID NO:86 MHLTLFTVAVLLLAAAALLLLLPPAYSTTLTPPAP

SEQ ID NO:87 MPLSYCHLFLTHTLARALSFSRSDCLKF

SEQ ID NO:88 MKNVFFALLLVVLVCCLVSVQGNEIIQ

SEQ ID NO:89 GIKLLVLLICLFFYHTHCTTAYLWLAMGVEA

SEQ ID NO:90 MFLKGSFPRFQMCVMLIGFFSSAKCLMC

SEQ ID NO:91 KQNDYQALLGLCCPWIDLAAADLPMRRHAKARE

SEQ ID NO:92 MKFYSVGKLVKVLLVMAVCCLLLCTAPTGADPLPG

SEQ ID NO:93 MKFAFAFVLIALFAVFAVSQALPQPEQAAASS

SEQ ID NO:94 SNDGASAITKIVLELTPEQAAAVQK

SEQ ID NO:95 AITLFIFLVCCQIPLFGIMSSDSADPFYWIRVILASN

SEQ ID NO:96 MGRVMCLLRLMSTLLVVLSIVGKKT

SEQ ID NO:97 MKLYSGYRLLVLLVMTVCCLLLFIAPTGADPLPGQTQRTLGY

SEQ ID NO:98 CKMYCVIKGKTGGYCNSEGLCTCRAEDLHFLLKPIINKD

SEQ ID NO:99 TVNAEDPRTELIGCGSVLFHLAANRLSLQLEEFAVCKRSN

SEQ ID NO:100 CVALIGLLLCSVQSVTANDPVDALGACSGNLFGLLMTRLQQ

SEQ ID NO:101 MASKLFVLAFLCLALVVVVQSAPQYARGDVPTYD

SEQ ID NO:102 MKSMLVAFATLSVALVVVVAIPANFNYGGGGGYFINGTGQSF

SEQ ID NO:103 NAIYEKLPAYLSEVSARVNVLQVSLQHDLPNLQE

SEQ ID NO:104 STEMKLAKVALVTISLWFMAWTPYLVINFTGIFK

SEQ ID NO:105 GDLLPAKVIPDKTAAYVAYGGQETLVEHVEVLVHK

SEQ ID NO:106 NTFYTCFLGTSSLAGFKNAVDYDELLKAGKM

SEQ ID NO:107 QVLEVLGFVEDNGELVFQELLGVLKMVDPDGDHA

SEQ ID NO:108 RRKLTPTVVVVLLCLTFVADALTIQELRAQIAQQRIQQRYGVTVATTSA

SEQ ID NO:109 CSSLSDYGLIELKEHCLECCQKDTEADSKLKVYPAAVLEVCT

SEQ ID NO:110 AITYICFILHGVSEIIPQQQKKTMKFLLLVASVLCLVLIVS

SEQ ID NO:111 EWRYFVVIALICPLIIVETLAVSD

SEQ ID NO:112 QLLLLYLDAADLRRALHQYQLLAAQGDRHLPQQIVKFVYA

SEQ ID NO:113 RKVLLTPALQAYIMDEHNLNRSNIALGRIRPYPSAVKMPTL

SEQ ID NO:114 DGVLKGETHKALKLKDGGHYLVEFKSIYMAK

SEQ ID NO:115 ALVLHSMLVNASLAEMVKESYQTHGADGRMVVRMLKFVRLLPGA

SEQ ID NO:116 VQRVRALRALLETLLQHQGEQNNDVYLIRLAHETGR

SEQ ID NO:117 VNELQQALSSLNAGSGSCAEVFNAYLPVHNKYIGVSRKIYH

SEQ ID NO:118 MCKFYRLISTLLVVVVIAPRHQCSPFFFQYNRPYLSQ

SEQ ID NO:119 RDNYVPDVSALEQDIIEVDPETKEMLKHLDFNNIVVQLTN

SEQ ID NO:120 HDQYSMECLEAAEPKYLDGLKTLADETAQCMR

SEQ ID NO:121 EQEYAQVTKMLGNGRLEAMCFDGVKRLCHIRGKLRK

SEQ ID NO:122 MKLFLTLLSTLSVAMVFALPAHHHSRGGD

SEQ ID NO:123 SDELEEARLVAEELEERQQELDYLKRYLVGRLQAVAI

SEQ ID NO:124 IISYFLTVCLLALVQSETVQDCE

SEQ ID NO:125 DSAMTNANLVGLTISLAYAIFFLLYTPPTGRSSYW

SEQ ID NO:126 VVSFAWLLYGIILRSNFLVVQNLMALALSAVQLSLFIIFP

SEQ ID NO:127 TSAFPFISGFLSCFMWLKYGVLTEESTLILVNFIGSALFF

SEQ ID NO:128 VIGLLCCCLAVLFFASPLTMLAHVIRTQ

SEQ ID NO:129 VELLLAMVLLPLLLLESVVPYAAAEKVWVD

Such particularly preferred peptides include the following balances:

residues 1-35 >gi|18389913|gb|AAL68793.1|AF457563_1 hypothetical protein 16 [Anopheles gambiae]

residues 1-28 >gi|18389909|gb|AAL68791.1|AF457561_1 hypothetical protein 14 [Anopheles gambiae]

residues 1-27 >gi|18389907|gb|AAL68790.1|AF457560_1 hypothetical protein 13 [Anopheles gambiae]

the remains 8-38 >gi|18389903|gb|AAL68788.1|AF457558_1 hypothetical protein 11 [Anopheles gambiae]

residues 1-28 >gi|62546227|gb|AAX86005.1|hyp3.5-precursor [Anopheles gambiae]

the remains 12-44 gi|18389899|gb|AAL68786.1|AF457556_1 7-like protein in the salivary glands [Anopheles gambiae]

residues 1-35 >gi|18389911|gb|AAL68792.1|AF457562_1 hypothetical protein 15 [Anopheles gambiae]

residues 132 > gi|62546225|gb|AAX86004.1|hyp6.3-precursor [Anopheles gambiae]

the remains of 32-56 >gi|62546225|gb|AAX86004.1|hyp6.3 - precursor [Anopheles gambiae]

the remains of 36-72 >gi|17026153|emb|CAD12038.1|Sec61 protein [Anopheles gambiae]

residues 1-25 >gi|62546223|gb|AAX86003.1|hyp6.2-precursor [Anopheles gambiae]

the remains 15-56 >gi|18389915|gb|AAL68794.1|AF457564_1 hypothetical protein 17 [Anopheles gambiae]

the remains 55-93 >gi|87080391|gb|ABD18596.1|defensin [Anopheles gambiae]

the remains of 20-59 >gi|18389901|gb|AAL68787.1|AF457557_1 hypothetical protein 10 [Anopheles gambiae]

the remains of 5-45 >gi|18389905|gb|AAL68789.1|AF457559_1 hypothetical protein 12 [Anopheles gambiae]

residues 1-34 >gi|4127344|emb|CAA76832.1|cE5-protein [Anopheles gambiae]

residues 1-42 >gi|4210617|emb|CAA10259.1|SG2 protein [Anopheles gambiae]

the remains 89-122 >gi|4127309|emb|CAA76820.1|hypothetical protein [Anopheles gambiae]

the remains 63-96 >gi|4375824|emb|CAA76825.1|opsin has been performed [Anopheles gambiae]

the remains of 39-73 >gi|62546233|gb|AAX86008.1|unknown protein [Anopheles gambiae]

the remains of 115-145 >gi|3378531|emb|CAA03872.1|D7r2 protein [Anopheles gambiae]

residues 61-95 >gi|3378529|emb|CAA03871.1|D7r3 protein [Anopheles gambiae]

the remains 21-69 >gi|18389893|gb|AAL68783.1|AF457553_1 muzykopodobnyh protein [Anopheles gambiae]

the remains 41-82 >gi|18389881|gb|AAL68777.1|AF457547_1 selenoprotein [Anopheles gambiae]

the remains 4-44 >gi|18389879|gb|AAL68776.1|AF457546_1 protein with Mm 30 kDa [Anopheles gambiae]

residues 2-25 >gi|18378603|gb|AAL68639.1|AF458073_1 D7-related protein 5 [Anopheles gambiae]

the remains of 18-57 >gi|18389897|gb|AAL68785.1|AF457555_1 1-like protein 4 salivary glands [Anopheles gambiae]

the remains 57-97 >gi|18389883|gb|AAL68778.1|AF457548_1 antigen 5-related proteins is 1 [Anopheles gambiae]

the remains 156-186 >gi|83016748|dbj|BAE53441.1|DsRed [synthetic constructs]

the remains 35-78 >gi|18389895|gb|AAL68784.1|AF457554_1 1-like protein 3 salivary glands [Anopheles gambiae]

the remains 189-224 >gi|18389895|gb|AAL68784.1|AF457554_1 1-like protein 3 salivary glands [Anopheles gambiae]

the remains 111-151 >gi|18389891|gb|AAL68782.1|AF457552_1 long form D7-protein [Anopheles gambiae]

residues 1-37 >emb|CAC35527.1|gSG9-protein [Anopheles gambiae]

the remains 81-120 >sp|Q9U9L1|RS17_ANOGA protein S17 40S subunit of the ribosome

the remains 111-142 >emb|CAC35523.1|gSG7 protein [Anopheles gambiae]

the remains 32-67 >gb|AAD47075.1|AF164151_1 factor 4C (1A) of translation initiation [Anopheles gambiae]

residues 1-29 >emb|CAC35519.1|gSG2-like protein [Anopheles gambiae]

the remains 106-142 >emb|CAC35451.1|hypothetical protein [Anopheles gambiae]

residues 6-28 >emb|CAC35524.1|D7r4 protein [Anopheles gambiae]

the remains 70-104 >ref|XP_001230998.1|ENSANGP00000014906 [Anopheles gambiae strain PEST]

the remains 174-213 >ref|XP_316361.2|ENSANGP00000012984 [Anopheles gambiae strain PEST]

the remains 41-80 >ref|XP_314140.3|ENSANGP00000015780 [Anopheles gambiae strain PEST]

the remains 126-153 >ref|XP_314140.3|ENSANGP00000015780 [Anopheles gambiae strain PEST]

residues 5-34 >emb|CAC35522.1|gSG6 protein [Anopheles gambiae]

In alternative embodiments, the implementation of the present invention it is directed to compositions containing polypeptides that are homologous to the above sequences, in particular peptides that are homologous to any of SEQ ID NO:1-134. Homology referred to above in respect of these sequences is predpochtitelno%, 75%, 80%, 85%, 90%, 95% or essentially 100%.

The homology percentage of the first sequence of the second polypeptide sequence of the polypeptide referred to in the context of the present invention, is defined as the number of amino acid residues in the second sequence that match both the position and identity of amino acid residues in the first sequence, divided by the total number of amino acid residues of the second polypeptide (first and second polypeptides must have the same number of amino acid residues) and multiplied by 100. In the present invention, preferably, when the homology of the polypeptide defined above sequence is 75% or more, 80% or more, 85% or more, 90% or more, 95% or more or 100% (or essentially 100%).

In the present invention redistributable arthropod disease is not particularly limited, and polypeptides can be immunogenic in respect of any known distributed arthropod disease and/or derived from it. Examples of diseases, pathogens and disease vectors covered by the present invention described above in table 1. However, preferably the relevant disease is malaria (including any malarial strain)caused by any of the strains of plasmodia.

Specific sequences, homola is similar to any of SEQ ID NO:1-134, defined above, are preferably sequences in the relevant provisions in known proteins arthropods that can be found in the database for proteins common use NCBI, which you can obtain at the following URL: http://www.ncbi.nlm.nih.gov/entrez/query/static/help/helpdoc.html#Protein. The list is typically presented in the form of |version number (gi number)|the ID of the database (e.g., gb for GenBank)|incoming number in NCBI|optional additional information (for example, incoming number the nucleotide sequence from which is deduced sequence of the protein). Protein database contains sequence data derived from the translated coding regions of DNA sequences in GenBank, EMBL and DDBJ, as well as sequences of proteins represented in the protein Information resource (PIR), SwissProt, the Foundation for the study of proteins (PRF) and the data Bank of protein structure (PDB) (sequences from solved structures).

Epitopes within the above defined sequences are not particularly limited provided that they contain 7 amino acids or more. Preferably, the epitopes are at least the length that is appropriate for the least immunogenic epitopes, such as CTL epitopes, T-helper cells and b-cells in the spine, such as man. Typically, epitopes will win 8, 9, 10, or 11 amino acid residues, but may contain more amino acid residues, if desired.

Although the polypeptide can include more amino acids, as a rule, it contains 100 amino acids or less, preferably between 7 and 100 amino acids, and more preferably 8-75 amino acids. The size should not be so large that suitable epitopes may suffer from competition with aprotective the epitopes in the immune system (for this reason, a full-sized proteins not included), the size should not be so small that only a very narrow range of protection is provided. More preferred ranges are 15-75 amino acids, 20-55 amino acids and 23-50 amino acids. Particularly preferably, when the polypeptide is (or essentially consists of a sequence selected from the above defined sequences.

In addition to the above polypeptides of the present invention is also provided polyepitope immunogenic polypeptides containing two or more polypeptides of the present invention or in the form of a repeatedly branched polypeptides, or connected in a chain sequence. The size of these polyepitope polypeptide is not limited and may be, for example, up to 1400, or up to 900, or up to 550 amino acids. Thus, they apply not only n is defined above polypeptides, but also for large polypeptides, provided that these large polypeptides include two or more units, each unit consists of a polypeptide of the present invention. Therefore, the present invention is covered by a polypeptide having 100 recurring 7-dimensional units in accordance with the present invention, as a polypeptide having, say, 52 units of one 8-dimensional epitope and 23 units of the second 10-dimensional epitope. The polypeptides of this type will not suffer the problems associated with competition with polypeptides of similar length, which include only one or two epitope. For the avoidance of doubt polyepitope polypeptide can include multiple copies of the same epitope, or single copies of a number of different epitopes, or multiple copies of 2 or more epitopes. Particularly preferably, when polyepitope polypeptide contains two or more sequences described in SEQ ID NO:1-44 (and especially the sequences in SEQ ID NO:1-6, 7, 8, 11, 12, 15, 16, 20, 26, 28, 30-32, 35, 41 and 42) or in SEQ ID NO:86-134.

As already mentioned, the present invention provides polypeptide composition comprising one or more, preferably two or more different polypeptides, as defined above. Thus, the polypeptide composition may include any number of polypeptides of the present invention along the same sequence, the mixture or composition. The presence together of many polypeptides useful, because each of them can induce its own immune response, expanding the protective effect of the composition. Particularly preferably, when the composition contains two or more (or all) of the sequences SEQ ID NO:1-44 (and especially the sequences in SEQ ID NO:1-6, 7, 8, 11, 12, 15, 16, 20, 26, 28, 30-32, 35, 41 and 42) and/or two or more epitopes within SEQ ID NO:86-134. The composition of each sequence and/or the epitope may be present or in isolated peptide, or in the form of a number of larger peptides comprising multiple connected in the circuit of epitopes and/or sequences (for example, three sequences connected in the circuit of one larger peptide, and the other 4 sequences connected in the circuit of another larger peptide, and the like).

The present invention also provides polypeptide construct that includes the above-defined polypeptide and the media. The design can be created by combining one or more epitopes and/or polypeptides, as defined above, with a carrier. The media can be a molecule, such as an adjuvant and/or excipient. In this context, the Association means or mixing together, or linking together (for example, through a covalent bond).

The present invention also provide the correspondingly defined above polypeptide for use in medicine. Also provided drug or vaccine composition against distributed arthropod diseases, comprising the above-defined polypeptide and one or more suitable excipients and/or adjuvants or polypeptide defined above design and optional one or more suitable excipients and/or adjuvants (if which is the carrier of the structure itself is an excipient or adjuvant, an additional excipient or adjuvant may not be needed). The excipient or adjuvant is not particularly limited, and can be used any excipient or adjuvant used in medicines and vaccines. Drug or vaccine composition can be created in accordance with any known method, suitably adapted to the present invention, for example by mixing the polypeptide of the present invention with a suitable filler.

The present invention also provides a method of obtaining defined above polypeptide. The method is not particularly limited and usually involves a join of two or more epitopes with the formation of the polypeptide. The polypeptide can, however, be synthesized by direct chemical synthesis (for example, the inclusion of one amino acid at a time before the formation of Polarstern the second polypeptide) or recombinant methods. Such General methods well known qualified and can be adapted to the present invention, if desired. In some cases, the polypeptide of the present invention may include additional amino acid sequences at one or both ends to facilitate the synthesis of the polypeptide. Preferably the length of these additional sequences is 1-5 amino acids. Usually consist of 2 amino acids. Examples of such sequences are provided as SEQ ID NO:86-129 described above.

Still further, the present invention provides the use of certain of the above polypeptide or composition for the manufacture of a medicinal product or a vaccine that is effective for the treatment or prophylaxis of disseminated arthropod disease. Also provided is a method of treatment or prophylaxis of disseminated arthropod disease involving the introduction of a vertebral defined above polypeptide, composition, medicament or vaccine. The route of administration is not particularly limited and may include subcutaneous, intramuscular, intravenous, intradermal or intranasal introduction, or oral administration (for example, in the form of tablets or liquid preparation), or the introduction in the form of a suppository, if desirable. The form of preparations for such introductions special is not limited to, and can be used a known form with appropriate modifications that will be obvious to a qualified specialist. The dose is not particularly limited and may be in the range from 1 PG to 100 g, preferably from 1 ng to 100 g of the polypeptide for each individual, depending on size, weight and form of the involved individual.

The present invention is applicable to any vertebral because the immune system of vertebrates are functioning properly. Usually mentioned in the context of the present invention vertebrate is a mammal, bird, reptile or fish. Particularly preferably, when the vertebrate is a human, a domestic animal (such as a dog or cat), farm animals (such as pig or horse), cattle (for example, cows and bulls) or poultry (e.g., poultry, agricultural bird or bird fighting breed). If the vertebrate is a bird, it preferably is chicken, Turkey, duck or goose.

Examples of human MHC (HLA), which may contact a specific T-cell epitope in the present invention include the following MHC (HLA):

HLA-A

A*010101, A*010102, A*010103, A*0102, A*0103, A*0104N, A*0106, A*0107, A*0108, A*0109, A*0110, A*02010101, A*02010102L, A*020102, A*020103, A*020104, A*020105, A*020106, A*020107, A*020108, A*020109, A*020110, A*020111, A*0202, A*020301 A*020302, A*0204, A*0205, A*020601, A*020602, A*020603, A*0207, A*0208, A*0209, A*0210, A*0211, A*0212, A*0213, A*0214, A*0215N, A*0216, A*021701, A*021702, A*0218, A*0219, A*022001, A*022002, A*0221, A*0222, A*0224, A*0225, A*0226, A*0227, A*0228, A*0229, A*0230, A*0231, A*0232N, A*0233, A*0234, A*023501, A*023502, A*0236, A*0237, A*0238, A*0239, A*0240, A*0241, A*0242, A*0243N, A*0244, A*0245, A*0246, A*0247, A*0248, A*0249, A*0250, A*0251, A*0252, A*0253N, A*0254, A*0255, A*0256, A*0257, A*0258, A*0259, A*0260, A*0261, A*0262, A*0263, A*0264, A*0265, A*0266, A*0267, A*0268, A*0269, A*0270, A*0271, A*0272, A*0273, A*03010101, A*03010102N, A*03010103, A*030102, A*030103, A*0302, A*0303N, A*0304, A*0305, A*0306, A*0307, A*0308, A*0309, A*0310, A*0311N, A*0312, A*0313, A*0314, A*110101, A*110102, A*1102, A*1103, A*1104, A*1105, A*1106, A*1107, A*1108, A*1109, A*1110, A*1111, A*1112, A*1113, A*1114, A*1115, A*1116, A*1117, A*1118, A*1119, A*2301, A*2302, A*2303, A*2304, A*2305, A*2306, A*2307N, A*2308N, A*2309, A*2310, A*2311N, A*2312, A*24020101, A*24020102L, A*240202, A*240203, A*240204, A*240205, A*240206, A*240301, A*240302, A*2404, A*2405, A*2406, A*2407, A*2408, A*2409N, A*2410, A*2411N, A*2413, A*2414, A*2415, A*2417, A*2418, A*2419, A*2420, A*2421, A*2422, A*2423, A*2424, A*2425, A*2426, A*2427, A*2428, A*2429, A*2430, A*2431, A*2432, A*2433, A*2434, A*2435, A*2436N, A*2437, A*2438, A*2439, A*2440N, A*2441, A*2442, A*2443, A*2444, A*2445N, A*2446, A*250101, A*250102, A*2502, A*2503, A*2504, A*2601, A*2602, A*2603, A*2604, A*2605, A*2606, A*260701, A*260702, A*2608, A*2609, A*2610, A*2611N, A*2612, A*2613, A*2614, A*2615, A*2616, A*2617, A*2618, A*2619, A*2620, A*2621, A*2622, A*2623, A*29010101, A*29010102N, A*290201, A*290202, A*290203, A*2903, A*2904, A*2905, A*2906, A*2907, A*2908N, A*2909, A*2910, A*2911, A*300101, A*300102, A*300201, A*300202, A*3003, A*3004, A*3006, A*3007, A*3008, A*3009, A*3010, A*3011, A*3012, A*310102, A*3102, A*3103, A*3104, A*3105, A*3106, A*3107, A*3108, A*3109, A*3110, A*3201, A*3202, A*3203, A*3204, A*3205, A*3206, A*3207, A*3208, A*3301, A*330301, A*330302, A*3304, A*3305, A*3306, A*3307, A*3401, A*3402, A*3403, A*3404, A*3405, A*3406, A*3601, A*3602, A*3603, A*3604, A*4301, A*6601, A*6602, A*6603, A*6604, A*680101, A*680102, A*680103, A*6802, A*680301, A*680302, A*6804, A*6805, A*6806, A*6807, A*6808, A*6809, A*6810, A*6811N, A*812, A*6813, A*6814, A*6815, A*6816, A*6817, A*6818N, A*6819, A*6820, A*OF 6,821, A*6822, A*6823, A*OF 6,824, A*6825, A*6826, A*6827, A*6901, A*7401, A*7402, A*7403, A*7404, A*7405, A*7406, A*7407, A*7408, A*7409, A*7410, A*8001.

HLA-B

B*070201, B*070202, B*070203, B*070204, B*0703, B*0704, B*0705, B*0706, B*0707, B*0708, B*0709, B*0710, B*0711, B*0712, B*0713, B*0714, B*0715, B*0716, B*0717, B*0718, B*0719, B*0720, B*0721, B*0722, B*0723, B*0724, B*0725, B*0726, B*0727, B*0728, B*0729, B*0730, B*0731, B*0732, B*0733, B*0734, B*0735, B*0736, B*0737, B*0738, B*0801, B*0802, B*0803, B*0804, B*0805, B*0806, B*0807, B*0808N, B*0809, B*0810, B*0811, B*0812, B*0813, B*0814, B*0815, B*0816, B*0817, B*0818, B*0819N, B*0820, B*0821, B*0822, B*1301, B*1302, B*1303, B*1304, B*1306, B*1307N, B*1308, B*1309, B*1310, B*1311, B*1312, B*1313, B*1401, B*1402, B*1403, B*1404, B*1405, B*140601, B*140602, B*15010101, B*15010102N, B*150102, B*150103, B*150104, B*150105, B*1502, B*1503, B*1504, B*1505, B*1506, B*1507, B*1508, B*1509, B*1510, B*151101, B*151102, B*1512, B*1513, B*1514, B*1515, B*1516, B*15170101, B*15170102, B*1518, B*1519, B*1520, B*1521, B*1523, B*1524, B*1525, B*1526N, B*1527, B*1528, B*1529, B*1530, B*1531, B*1532, B*1533, B*1534, B*1535, B*1536, B*1537, B*1538, B*1539, B*1540, B*1542, B*1543, B*1544, B*1545, B*1546, B*1547, B*1548, B*1549, B*1550, B*1551, B*1552, B*1553, B*1554, B*1555, B*1556, B*1557, B*1558, B*1560, B*1561, B*1562, B*1563, B*1564, B*1565, B*1566, B*1567, B*1568, B*1569, B*1570, B*1571, B*1572, B*1573, B*1574, B*1575, B*1576, B*1577, B*1578, B*1579N, B*1580, B*1581, B*1582, B*1583, B*1584, B*1585, B*1586, B*1587, B*1588, B*1589, B*1590, B*1591, B*1592, B*1593, B*1594N, B*180101, B*180102, B*1802, B*1803, B*1804, B*1805, B*1806, B*1807, B*1808, B*1809, B*1810, B*1811, B*1812, B*1813, B*1814, B*1815, B*1817N, B*1818, B*1819, B*1820, B*2701, B*2702, B*2703, B*2704, B*270502, B*270503, B*270504, B*270505, B*270506, B*270507, B*2706, B*2707, B*2708, B*2709, B*2710, B*2711, B*2712, B*2713, B*2714, B*2715, B*2716, B*2717, B*2718, B*2719, B*2720, B*2721, B*2723, B*2724, B*2725, B*2726, B*350101, B*350102, B*3502, B*3503, B*3504, B*3505, B*3506, B*3507, B*3508, B*350901, B*350902, B*3510, B*3511, B*3512, B*3513, B*351401, B*351402, B*3515, B*3516, B*3517, B*3518, B*3519, B*3520, B*3521, B*3522, B*3523, B*3524, B*3525, B*3526, B*3527, B*358, B*3529, B*3530, B*3531, B*3532, B*3533, B*3534, B*3535, B*3536, B*3537, B*3538, B*3539, B*3540N, B*3541, B*3542, B*3543, B*3544, B*3545, B*3546, B*3547, B*3548, B*3549, B*3550, B*3551, B*3552, B*3553N, B*3701, B*3702, B*3703N, B*3704, B*3705, B*3706, B*3707, B*3801, B*380201, B*380202, B*3803, B*3804, B*3805, B*3806, B*3807, B*3808, B*3809, B*3810, B*390101, B*390103, B*390104, B*390201, B*390202, B*3903, B*3904, B*3905, B*390601, B*390602, B*3907, B*3908, B*3909, B*3910, B*3911, B*3912, B*3913, B*3914, B*3915, B*3916, B*3917, B*3918, B*3919, B*3920, B*3922, B*3923, B*3924, B*3925N, B*3926, B*3927, B*3928, B*3929, B*3930, B*3931, B*3932, B*400101, B*400102, B*400103, B*400104, B*400105, B*400201, B*400202, B*4003, B*4004, B*4005, B*40060101, B*40060102, B*4007, B*4008, B*4009, B*4010, B*4011, B*4012, B*4013, B*401401, B*401402, B*401403, B*4015, B*4016, B*4018, B*4019, B*4020, B*4021, B*4022N, B*4023, B*4024, B*4025, B*4026, B*4027, B*4028, B*4029, B*4030, B*4031, B*4032, B*4033, B*4034, B*4035, B*4036, B*4037, B*4038, B*4039, B*4040, B*4042, B*4043, B*4044, B*4045, B*4046, B*4047, B*4048, B*4049, B*4050, B*4051, B*4052, B*4053, B*4054, B*4055, B*4056, B*4057, B*4101, B*4102, B*4103, B*4104, B*4105, B*4106, B*4201, B*4202, B*4204, B*420501, B*420502, B*4206, B*44020101, B*44020102S, B*440202, B*440203, B*440301, B*440302, B*4404, B*4405, B*4406, B*4407, B*4408, B*4409, B*4410, B*4411, B*4412, B*4413, B*4414, B*4415, B*4416, B*4417, B*4418, B*4419N, B*4420, B*4421, B*4422, B*4423N, B*4424, B*4425, B*4426, B*4427, B*4428, B*4429, B*4430, B*4431, B*4432, B*4433, B*4434, B*4435, B*4436, B*4437, B*4438, B*4439, B*4440, B*4501, B*4502, B*4503, B*4504, B*4505, B*4506, B*4507, B*4601, B*4602, B*4603, B*4604, B*47010101, B*47010102, B*4702, B*4703, B*4704, B*4705, B*4801, B*4802, B*4803, B*4804, B*4805, B*4806, B*4807, B*4808, B*4809, B*4810, B*4901, B*4902, B*4903, B*5001, B*5002, B*5004, B*510101, B*510102, B*510103, B*510104, B*510105, B*510201, B*510202, B*5103, B*5104, B*5105, B*5106, B*5107, B*5108, B*5109, B*5110, B*5111N, B*5112, B*511301, B*511302, B*5114, B*5115, B*5116, B*5117, B*5118, B*5119, B*5120, B*5121, B*5122, B*5123, B*5124, B*5126, B*5127N, B*5128, B*5129, B*5130, B*5131, B*5132, B*5133, B*5134, B*5135, B*5136, B*520101, B*520102, B*520103, B*520104, B*5202, B*5203, B*5204, B*5205, B*5206, B*530101, B*530102, B*5302, B*5303, B*5304, B*5305, B*5306 B*5307, B*5308, B*5309, B*5401, B*5402, B*5501, B*5502, B*5503, B*5504, B*5505, B*5507, B*5508, B*5509, B*5510, B*5511, B*5512, B*5513, B*5514, B*5515, B*5516, B*5601, B*5602, B*5603, B*5604, B*560501, B*560502, B*5606, B*5607, B*5608, B*5609, B*5610, B*5611, B*5612, B*5613, B*5614, B*570101, B*570102, B*5702, B*570301, B*570302, B*5704, B*5705, B*5706, B*5707, B*5708, B*5709, B*5801, B*5802, B*5804, B*5805, B*5806, B*5807, B*5808, B*5809, B*5810N, B*5901, B*670101, B*670102, B*6702, B*7301, B*7801, B*780201, B*780202, B*7803, B*7804, B*7805, B*8101, B*8102, B*8201, B*8202, B*8301.

HLA-C

Cw*010201, Cw*010202, Cw*0103, Cw*0104, Cw*0105, Cw*0106, Cw*0107, Cw*0108, Cw*0109, Cw*0110, Cw*020201, Cw*020202, Cw*020203, Cw*020204, Cw*020205, Cw*0203, Cw*0204, Cw*0205, Cw*0206, Cw*0207, Cw*0208, Cw*0209, Cw*030201, Cw*030202, Cw*030301, Cw*030302, Cw*030303, Cw*030304, Cw*030401, Cw*030402, Cw*030403, Cw*0305, Cw*0306, Cw*0307, Cw*0308, Cw*0309, Cw*0310, Cw*0311, Cw*0312, Cw*0313, Cw*0314, Cw*0315, Cw*0316, Cw*0317, Cw*0318, Cw*04010101, Cw*04010102, Cw*040102, Cw*0403, Cw*040401, Cw*040402, Cw*0405, Cw*0406, Cw*0407, Cw*0408, Cw*0409N, Cw*0410, Cw*0411, Cw*0412, Cw*0413, Cw*0414, Cw*0415, Cw*050101, Cw*050102, Cw*0502, Cw*0503, Cw*0504, Cw*0505, Cw*0506, Cw*0507N, Cw*0508, Cw*0509, Cw*0510, Cw*0602, Cw*0603, Cw*0604, Cw*0605, Cw*0606, Cw*0607, Cw*0608, Cw*0609, Cw*0610, Cw*0611, Cw*070101, Cw*070102, Cw*070103, Cw*07020101, Cw*07020102, Cw*07020103, Cw*0703, Cw*070401, Cw*070402, Cw*0705, Cw*0706, Cw*0707, Cw*0708, Cw*0709, Cw*0710, Cw*0711, Cw*0712, Cw*0713, Cw*0714, Cw*0715, Cw*0716, Cw*0717, Cw*0718, Cw*0719, Cw*0720, Cw*0721, Cw*0722, Cw*0723, Cw*0724, Cw*0725, Cw*0726, Cw*0727, Cw*0728, Cw*0729, Cw*080101, Cw*080102, Cw*0802, Cw*0803, Cw*0804, Cw*0805, Cw*0806, Cw*0807, Cw*0808, Cw*0809, Cw*0810, Cw*0811, Cw*0812, Cw*120201, Cw*120202, Cw*120203, Cw*120301, Cw*120302, Cw*120303, Cw*120401, Cw*120402, Cw*1205, Cw*1206, Cw*1207, Cw*1208, Cw*1209, Cw*1210, Cw*1211, Cw*1212, Cw*1213, Cw*1214, Cw*1215, Cw*140201, Cw*140202, Cw*140203, Cw*1403, Cw*1404, Cw*1405, Cw*150201, Cw*150202, Cw*1503, Cw*1504, Cw*150501, Cw*150502, Cw*150503, Cw*150504, Cw*1506, Cw*1507, Cw*1508, Cw*1509, Cw*1510, Cw*1511, Cw*1512, Cw*1601, Cw*1602, Cw*160401, Cw*1606, Cw*1701, Cw*1702, Cw*1703, Cw*1801, Cw*1802.

HLA-E

E*0101, E*010301, E*010302, E*01303, E*0104.

HLA-F

F*010101, F*010102.

HLA-G

G*010101, G*010102, G*010103, G*010104, G*010105, G*010106, G*010107, G*010108, G*0102, G*0103, G*010401, G*010402, G*010403, G*0105N, G*0106.

HLA-DRA

DRA*0101, DRA*010201, DRA*010202.

HLA-DRB1

DRB1*010101, DRB1*010102, DRB1*010103, DRB1*010201, DRB1*010202, DRB1*010203, DRB1*010204, DRB1*0103, DRB1*0104, DRB1*0105, DRB1*0106, DRB1*0107, DRB1*0108, DRB1*0109, DRB1*0110, DRB1*0111, DRB1*030101, DRB1*030102, DRB1*030201, DRB1*030202, DRB1*0303, DRB1*0304, DRB1*030501, DRB1*030502, DRB1*0306, DRB1*0307, DRB1*0308, DRB1*0309, DRB1*0310, DRB1*0311, DRB1*0312, DRB1*0313, DRB1*0314, DRB1*0315, DRB1*0316, DRB1*0317, DRB1*0318, DRB1*0319, DRB1*0320, DRB1*0321, DRB1*0322, DRB1*0323, DRB1*0324, DRB1*0325, DRB1*0326, DRB1*0327, DRB1*0328, DRB1*040101, DRB1*040102, DRB1*0402, DRB1*040301, DRB1*040302, DRB1*0404, DRB1*040501, DRB1*040502, DRB1*040503, DRB1*040504, DRB1*0406, DRB1*040701, DRB1*040702, DRB1*040703, DRB1*0408, DRB1*0409, DRB1*0410, DRB1*0411, DRB1*0412, DRB1*0413, DRB1*0414, DRB1*0415, DRB1*0416, DRB1*0417, DRB1*0418, DRB1*0419, DRB1*0420, DRB1*0421, DRB1*0422, DRB1*0423, DRB1*0424, DRB1*0425, DRB1*0426, DRB1*0427, DRB1*0428, DRB1*0429, DRB1*0430, DRB1*0431, DRB1*0432, DRB1*0433, DRB1*0434, DRB1*0435, DRB1*0436, DRB1*0437, DRB1*0438, DRB1*0439, DRB1*0440, DRB1*0441, DRB1*0442, DRB1*0443, DRB1*0444, DRB1*0445, DRB1*0446, DRB1*0447, DRB1*0448, DRB1*0449, DRB1*0450, DRB1*070101, DRB1*070102, DRB1*0703, DRB1*0704, DRB1*0705, DRB1*0706, DRB1*0707, DRB1*0708, DRB1*080101, DRB1*080102, DRB1*080201, DRB1*080202, DRB1*080203, DRB1*080302, DRB1*080401, DRB1*080402, DRB1*080403, DRB1*080404, DRB1*0805, DRB1*0806, DRB1*0807, DRB1*0808, DRB1*0809, DRB1*0810, DRB1*0811, DRB1*0812, DRB1*0813, DRB1*0814, DRB1*0815, DRB1*0816, DRB1*0817, DRB1*0818, DRB1*0819, DRB1*0820, DRB1*0821, DRB1*0822, DRB1*0823, DRB1*0824, DRB1*0825, DRB1*0826, DRB1*0827, DRB1*0828, DRB1*0829, DRB1*090102, DRB1*090103, DRB1*0902, DRB1*0903, DRB1*100101, DRB1*100102, DRB1*110101, DRB1*110102, DRB1*110103, DRB1*110104, DRB1*110105, DRB1*1102, DRB1*1103, DRB1*110401, DRB1*110402, DRB1*1105, DRB1*110601, DRB1*110602, DRB1*1107, DRB1*110801, DRB1*110802, DRB1*1109, DRB1*1110, DRB1*1111, DRB1*111201, DRB1*111202, DRB1*1113, DRB1*1114, DRB1*1115, DRB1*1116, DRB1*1117, DR1*1118, DRB1*1119, DRB1*1120, DRB1*1121, DRB1*1122, DRB1*1123, DRB1*1124, DRB1*1125, DRB1*1126, DRB1*112701, DRB1*112702, DRB1*1128, DRB1*1129, DRB1*1130, DRB1*1131, DRB1*1132, DRB1*1133, DRB1*1134, DRB1*1135, DRB1*1136, DRB1*1137, DRB1*1138, DRB1*1139, DRB1*1140, DRB1*1141, DRB1*1142, DRB1*1143, DRB1*1144, DRB1*1145, DRB1*1146, DRB1*1147, DRB1*1148, DRB1*1149, DRB1*1150, DRB1*1151, DRB1*1152, DRB1*1153, DRB1*1154, DRB1*120101, DRB1*120102, DRB1*120201, DRB1*120202, DRB1*120302, DRB1*1204, DRB1*1205, DRB1*1206, DRB1*1207, DRB1*1208, DRB1*1209, DRB1*1210, DRB1*130101, DRB1*130102, DRB1*130103, DRB1*130201, DRB1*130202, DRB1*130301, DRB1*130302, DRB1*1304, DRB1*1305, DRB1*1306, DRB1*130701, DRB1*130702, DRB1*1308, DRB1*1309, DRB1*1310, DRB1*1311, DRB1*1312, DRB1*1313, DRB1*131401, DRB1*131402, DRB1*1315, DRB1*1316, DRB1*1317, DRB1*1318, DRB1*1319, DRB1*1320, DRB1*1321, DRB1*1322, DRB1*1323, DRB1*1324, DRB1*1325, DRB1*1326, DRB1*1327, DRB1*1328, DRB1*1329, DRB1*1330, DRB1*1331, DRB1*1332, DRB1*1333, DRB1*1334, DRB1*1335, DRB1*1336, DRB1*1337, DRB1*1338, DRB1*1339, DRB1*1340, DRB1*1341, DRB1*1342, DRB1*1343, DRB1*1344, DRB1*1345, DRB1*1346, DRB1*1347, DRB1*1348, DRB1*1349, DRB1*1350, DRB1*1351, DRB1*1352, DRB1*1353, DRB1*1354, DRB1*1355, DRB1*1356, DRB1*1357, DRB1*1358, DRB1*1359, DRB1*1360, DRB1*1361, DRB1*1362, DRB1*1363, DRB1*1364, DRB1*1365, DRB1*140101, DRB1*140102, DRB1*1402, DRB1*140301, DRB1*140302, DRB1*1404, DRB1*140501, DRB1*140502, DRB1*1406, DRB1*140701, DRB1*140702, DRB1*1408, DRB1*1409, DRB1*1410, DRB1*1411, DRB1*1412, DRB1*1413, DRB1*1414, DRB1*1415, DRB1*1416, DRB1*1417, DRB1*1418, DRB1*1419, DRB1*1420, DRB1*1421, DRB1*1422, DRB1*1423, DRB1*1424, DRB1*1425, DRB1*1426, DRB1*1427, DRB1*1428, DRB1*1429, DRB1*1430, DRB1*1431, DRB1*1432, DRB1*1433, DRB1*1434, DRB1*1435, DRB1*1436, DRB1*1437, DRB1*1438, DRB1*1439, DRB1*1440, DRB1*1441, DRB1*1442, DRB1*1443, DRB1*1444, DRB1*1445, DRB1*1446, DRB1*1447, DRB1*1448, DRB1*150101, DRB1*150102, DRB1*150103, DRB1*150104, DRB1*150105, DRB1*150201, DRB1*150202, DRB1*150203, DRB1*1503, DRB1*1504, DRB1*1505, DRB1*1506, DRB1*1507, DRB1*1508, DRB1*1509, DRB1*1510, DRB1*1511, DRB1*1512, DRB1*1513, DRB1*1514, DRB1*1515, DRB1*1516, DRB1*160101, DRB1*160102, DRB1*160201, DRB1*160202, DRB1*1603, DRB1*1604, DRB1*160501, DRB1*160502, DRB1*1607, DRB1*1608.

HLA-DRB2-9

DRB2*0101, DRB3*010101, DRB3*01010201, DB3*01010202, DRB3*010103, DRB3*010104, DRB3*0102, DRB3*0103, DRB3*0104, DRB3*0105, DRB3*0106, DRB3*0107, DRB3*0108, DRB3*0109, DRB3*0110, DRB3*0111, DRB3*0201, DRB3*020201, DRB3*020202, DRB3*020203, DRB3*020204, DRB3*0203, DRB3*0204, DRB3*0205, DRB3*0206, DRB3*0207, DRB3*0208, DRB3*0209, DRB3*0210, DRB3*0211, DRB3*0212, DRB3*0213, DRB3*0214, DRB3*0215, DRB3*0216, DRB3*0217, DRB3*0218, DRB3*0219, DRB3*030101, DRB3*030102, DRB3*0302, DRB3*0303, DRB4*01010101, DRB4*0102, DRB4*01030101, DRB4*01030102N, DRB4*010302, DRB4*010303, DRB4*010304, DRB4*0104, DRB4*0105, DRB4*0106, DRB4*0107, DRB4*0201N, DRB4*0301N, DRB5*010101, DRB5*010102, DRB5*0102, DRB5*0103, DRB5*0104, DRB5*0105, DRB5*0106, DRB5*0107, DRB5*0108N, DRB5*0109, DRB5*0110N, DRB5*0111, DRB5*0112, DRB5*0113, DRB5*0202, DRB5*0203, DRB5*0204, DRB5*0205, DRB6*0101, DRB6*0201, DRB6*0202, DRB7*010101, DRB7*010102, DRB8*0101, DRB9*0101.

HLA-DQA1

DQA1*010101, DQA1*010102, DQA1*010201, DQA1*010202, DQA1*0103, DQA1*010401, DQA1*010402, DQA1*0105, DQA1*0106, DQA1*0107, DQA1*0201, DQA1*030101, DQA1*0302, DQA1*0303, DQA1*040101, DQA1*040102, DQA1*0402, DQA1*0403N, DQA1*0404, DQA1*050101, DQA1*050102, DQA1*0502, DQA1*0503, DQA1*0504, DQA1*0505, DQA1*060101, DQA1*060102, DQA1*0602.

HLA-DQB1

DQB1*020101, DQB1*020102, DQB1*0202, DQB1*0203, DQB1*030101, DQB1*030102, DQB1*030201, DQB1*030202, DQB1*030302, DQB1*030303, DQB1*0304, DQB1*030501, DQB1*030502, DQB1*030503, DQB1*0306, DQB1*0307, DQB1*0308, DQB1*0309, DQB1*0310, DQB1*0311, DQB1*0312, DQB1*0313, DQB1*0401, DQB1*0402, DQB1*050101, DQB1*050102, DQB1*050201, DQB1*050202, DQB1*050301, DQB1*050302, DQB1*0504, DQB1*060101, DQB1*060102, DQB1*060103, DQB1*0602, DQB1*0603, DQB1*060401, DQB1*060402, DQB1*060501, DQB1*060502, DQB1*0606, DQB1*0607, DQB1*0608, DQB1*0609, DQB1*0610, DQB1*061101, DQB1*061102, DQB1*0612, DQB1*0613, DQB1*0614, DQB1*0615, DQB1*0616, DQB1*0617, DQB1*0618, DQB1*0619, DQB1*0620, DQB1*0621, DQB1*0622, DQB1*0623.

HLA-DPA1

DPA1*010301, DPA1*010302, DPA1*010303, DPA1*0104, DPA1*0105, DPA1*0106, DPA1*0107, DPA1*0108, DPA1*020101, DPA1*020102, DPA1*020103, DPA1*020104, DPA1*020105, DPA1*020106, DPA1*020201, DPA1*020202, DPA1*020203, DPA1*0203, DPA1*0301, DPA1*0302, DPA1*0303, DPA1*0401.

HLA-DPB1

DPB1*010101, DPB1*010102, DPB1*010103, DPB1*0102, DPB1*020102, DPB1*020103, DPB*020104, DPB1*020105, DPB1*020106, DPB1*0202, DPB1*0203, DPB1*030101, DPB1*030102, DPB1*0302, DPB1*040101, DPB1*040102, DPB1*0402, DPB1*0501, DPB1*0601, DPB1*0801, DPB1*0901, DPB1*1001, DPB1*110101, DPB1*110102, DPB1*1301, DPB1*1401, DPB1*1501, DPB1*1601, DPB1*1701, DPB1*1801, DPB1*1901, DPB1*200101, DPB1*200102, DPB1*2101, DPB1*2201, DPB1*2301, DPB1*2401, DPB1*2501, DPB1*260101, DPB1*260102, DPB1*2701, DPB1*2801, DPB1*2901, DPB1*3001, DPB1*3101, DPB1*3201, DPB1*3301, DPB1*3401, DPB1*3501, DPB1*3601, DPB1*3701, DPB1*3801, DPB1*3901, DPB1*4001, DPB1*4101, DPB1*4401, DPB1*4501, DPB1*4601, DPB1*4701, DPB1*4801, DPB1*4901, DPB1*5001, DPB1*5101, DPB1*5201, DPB1*5301, DPB1*5401, DPB1*5501, DPB1*5601, DPB1*5701, DPB1*5801, DPB1*5901, DPB1*6001, DPB1*6101N, DPB1*6201, DPB1*6301, DPB1*6401N, DPB1*6501, DPB1*6601, DPB1*6701, DPB1*6801, DPB1*6901, DPB1*7001, DPB1*7101, DPB1*7201, DPB1*7301, DPB1*7401, DPB1*7501, DPB1*7601, DPB1*7701, DPB1*7801, DPB1*7901, DPB1*8001, DPB1*8101, DPB1*8201, DPB1*8301, DPB1*8401, DPB1*8501, DPB1*8601, DPB1*8701, DPB1*8801, DPB1*8901, DPB1*9001, DPB1*9101, DPB1*9201, DPB1*9301, DPB1*9401, DPB1*9501, DPB1*9601, DPB1*9701, DPB1*9801, DPB1*9901.

HLA-DMA

DMA*0101, DMA*0102, DMA*0103, DMA*0104.

HLA-DMB

DMB*0101, DMB*0102, DMB*0103, DMB*0104, DMB*0105, DMB*0106.

HLA-DOA

DOA*010101, DOA*01010201, DOA*01010202, DOA*01010203, DOA*010103, DOA*01010401, DOA*01010402, DOA*010105.

HLA-DOB

DOB*01010101, DOB*01010102, DOB*010102, DOB*010201, DOB*010202, DOB*0103, DOB*01040101, DOB*01040102.

MHC class I

H-2Db, H-2Dd, H-2Dk, H-2Dq, H-2Kb, H-2Kd and H-2Kk and H-2Ld, H-2M3, H-2Ad, H-2Ag7, H-2Ak, H2-Ab, H-2Ed, H-2Ek, H-2Bxk, H-2F, H-2I, H-2P, H-2R, H-2S, H-2Sxd, H-2T4, H-2U.

MHC class II

I-Ab, I-Ad, I-Ag7, I-Ak, I-Ap, I-Aq, I-Ar, I-As, I-Au I-Av, I-Ea, I-Eb, I-Ed, I-Ek, I-Es, I-Eu, H-2Q, H-2Qa-2, H-2Qa-2a, Qa-1, Qa-1b.

The present invention is not limited to such molecules MHC and HLA and can be adapted to newly discovered molecules, if desired, simply by creating the ability of substances, such as peptides, to respond to the Oia with these molecules. This can easily be successfully performed using known methods that are conventional in the art. Especially preferred for use with the present invention HLA alleles include the following alleles:

HLA class I

HLA-AHLA BHLA Cw
A*6802B*5801Cw*1701

A*6801B*5701Cw*1601
A*6601B*5501Cw*1502
A*3303B*5201Cw*1402
A*3301B*5101Cw*1203
A*3201B*5001Cw*0802
A*310102B*4901Cw*0801
A*3002B*4501Cw*0704
A*3001 B*4403Cw*0703
A*2902B*4402Cw*0702
A*2608B*4101Cw*0701
A*2601B*4002Cw*0602
A*2501B*4001Cw*0501
A*2402B*3901Cw*0401
A*2301B*3801Cw*0304
A*1101B*3701Cw*0303
A*0302B*3503Cw*0202
A*0301B*3501Cw*0102
A*0205B*2705
A*0201B*1801
A*0101B*1501
B*1402
B*1401
B*1302
B*0801
B*0705
B*0702

HLA class II

HLA DPBHLA DQAHLA DQBHLA DRB
DPB1*1701DQA1*0505DQB1*0604DRB1*1601
DPB1*1301DQA1*0501DQB1*0603DRB1*1501
DPB1*1001DQA1*0401DQB1*0602DRB1*1401
DPB1*0601DQA1*0303DQB1*0503DRB1*1302
DPB1*0501DQA1*0302 DQB1*0502DRB1*1301
DPB1*0402DQA1*0301DQB1*0501DRB1*1201
DPB1*0401DQA1*0201DQB1*0402DRB1*1104
DPB1*0301DQA1*0104DQB1*0303DRB1*1101
DPB1*0201DQA1*0103DQB1*0302DRB1*0801
DPB1*0101DQA1*0102DQB1*0301DRB1*0701
DQA1*0101DQB1*0202DRB1*0404
DQB1*0201DRB1*0401
DRB1*0301
DRB1*0103
DRB1*0102
DRB1*0101

The most preferred alleles in accordance with the present invention are the following alleles:

HLA-A*0201, HLA-A*0206, HLA-A*0301, HLA-A*1101, HLA-A*2402, HLA-A*3401, HLA-B*0702, HLA-B*0801, HLA-B*1301, HLA-B*27, HLA-B*4002, HLA-B*5101, HLA-Cw*03, HLA-cW*07

HLA-DRB1*0301, HLA-DRB1*0401, HLA-DRB1*0701, HLA-DRB1*1501, HLA-DRB1*1104, HLA-DRB1*1101, HLA-DRB4*0101

HLA-DQA1*01, HLA-DQA1*02, HLA-DQA1*05

HLA-DQB1*03, HLA-DQB1*04, HLA-DQB1*05, HLA-DQB1*06

HLA-DPA1*01, HLA-DPA1*02

HLA-DPB1*02, HLA-DPB1*04

The present invention will now be described only as an example, with reference to the following specific options for implementation.

EXAMPLES

Preparation of protein fractions of saliva arthropods

To determine the effect of specific sequences of the present invention their immunogenicity can be checked on the background of different protein fractions of arthropod saliva. Those sequences that cause spinal production of immune system cells that recognize at least one epitope specific protein fractions of saliva arthropods applicable in the vaccines of the present invention.

Protein fractions of saliva can easily be distinguished using standard laboratory methods, well known to the skilled Spa is ialist. You can use any protein fractions of arthropod saliva, because the authors of the present invention found that the really important weight protein fraction. In particular, the applicable fraction with a mass component 40 kDa or less, 30 kDa or less, preferably 20-40 kDa, and more preferably 20 kDa or less.

The following Protocol is provided for bringing the example of a protein fraction, against which you can test sequence candidates. It uses saliva from the mosquito Anopheles gambiae, although it can be used saliva of any arthropod.

A pair of salivary glands of Anopheles gambiae was dissected from the colonies of female mosquitoes. Fifteen pairs of salivary glands were collected in 20 μl PBS and literally by adding 5 μl of buffer for sample used for electrophoresis in SDS-page containing 0.25% 2-β-mercaptoethanol. After stirring and boiling for 5 min, the protein mixture was made in a 4-20%gradient Tris-glycine Novex gel (Invitrogen). Then the gel was stained with silver and photographed (see Fig.9).

The dotted arrows indicate the positions of the points cutoff for selection of fractions of pairs of salivary glands that can be used for immunization and research (ie <20 kDa, 20-40 kDa, 40-80 kDa and >80 kDa). The solid arrow indicates the position of the cutoff point for screening fractions of pairs of salivary glands, the components of <30 to the and and > 30 kDa.

Samples identification of protein candidates from the saliva of mosquitoes

Using an approach similar to that shown in example Protocol outlined above for the study was prepared several samples of proteins of saliva.

Salivary glands were dissected from female mosquitoes Anopheles gambiae and kept in PBS at -70°C until use.

For the analysis in gel salivary glands were literally freezing-thawing, was added to the buffer for sample (pH 3-10), used to perform IEF, Novex® (Invitrogen), and the resulting material was analyzed by gel IEF Novex® (Invitrogen). Then the gel was recorded using a 12% trichloroacetic acid, washed three times with water and stained Kumasi blue. The resulting gel is shown in figure 1.

After application and removal of the dye IEF gel was incubated in 20% ethanol for 10 minutes and the gel strip containing the line with proteins of the salivary glands, cut. This gel strip was balanced for 5 min in 2× buffer for sample used for electrophoresis in SDS-page, containing 20% ethanol, rinsed twice in buffer for sample used for electrophoresis in SDS-page, and made in one hole 4-20%gradient Tris-glycine gel (Novex® (Invitrogen). The resulting gel is shown in figure 2.

Finally, the gel containing separated b the CTL salivary glands, dyed using the dye Proteosilver (Sigma)according to manufacturer's instructions.

For the analysis identified four groups of proteins from the gel (named compounds 1, 2, 3 and 4):

Connection 1: protein fraction of the salivary glands by weight of <20 kDa

Connection 2: protein fraction of the salivary glands with a mass of 20 kDa < X < 40 kDa

Connection 3: protein fraction of the salivary glands with a mass of 40 kDa < X < 80 kDa

Compound 4: protein fraction of the salivary glands by weight of >80 kDa

EXPERIMENT 1

Objectives:

(A) to determine the effectiveness of the candidates in the protection of animals against infection by the bite of infected mosquitoes (i.e. Anopheles gambiae infected with Plasmodium yoelii nigeriensis).

(C) to Establish the level of cross-reactivity response against mosquitoes, induced by the candidates, for a set of different mosquito species (for example, Anopheles gambiae and Anopheles stephensi).

(C) to determine the efficacy of candidates in preventing infection of mosquitoes (Anopheles gambiae and Anopheles stephensi) agents Plasmodium yoelii nigeriensis bites immunized and infected mice.

Candidate connections

Selected compounds were defined above connections:

Connection 1: protein fraction of the salivary glands by weight of <20 kDa

Connection 2: protein fraction of the salivary glands with a mass of 20 kDa < X < 40 kDa

Connection 3: protein fraction of salivary the glands with a mass of 40 kDa < X < 80 kDa

Compound 4: protein fraction of the salivary glands by weight of >80 kDa

Strain and number of animals involved

Used CD1 mice. Used five experimental groups (1, 2, 3, 4 and 5), while group 1 was a negative control group, and groups 2-5 - subjects groups, with each group contained 9 animals.

Experimental Protocol

Day 1: 4 groups (groups 2, 3, 4 and 5) from 9 CD1 mice each (N=4×9=36) were immunized by subcutaneous injection of doses of candidate compounds to the vaccine (group 2 connection 1, group 3 connection 2 and so on).

Day 14: All mice were subjected to booster immunization with the same dose of the compounds-candidates for the vaccine (group 2 connection 1, group 3 connection 2 and so on).

Day 21: In all animals took blood samples. The samples were stored frozen (-20°C) prior to collection. Each group was further divided into subgroups: A - 5 animal - 4 animals.

Group 1A - 5 animalsGroup 1B - 4 animals
Group 2A - 5 animalsGroup 2B - 4 animals
Group 3A - 5 animalsGroup 3B - 4 animals
Group 4A - 5 alive is data Group 4B - 4 animals
Group 5A - 5 animalsGroup 5B - 4 animals

Subgroup a

Day 28: All animals in subgroups And were infected through bites 5-9 infected mosquitoes (i.e. Anopheles gambiae infected with Plasmodium yoelii nigeriensis) in the abdominal region. All animals were kept up to first establish the presence of parasites or for a maximum of 6 weeks after infection by infected mosquitoes. All animals were killed at the bleeding, and serum samples were stored frozen (-20°C) prior to collection.

Subgroup

Day 28: All animals in the subgroups used for feeding (in the abdomen) the following number of uninfected mosquitoes:

5-10 Anopheles gambiae and 5-10 Anopheles stephensi

All these mosquitoes were tested:

1. Survival during the 8-day period.

2. The number of eggs.

3. The number of produced eggs.

4. The number of adult mosquitoes (F)obtained from the eggs.

Day 32: All animals in subgroups In infected Plasmodium yoelii nigeriensis by direct intravenous injection of parasites.

Day 32: After the identification of active malaria infection in all (or at least 75%) of animals in the group In all infected animals used for feeding (in the abdomen) a large number (>10 for kardamyli) as uninfected Anopheles gambiaeand uninfectedAnopheles stephensi.

All these mosquitoes were tested:

1. Survival during the 8-day period.

2. The number of mosquitoes with malaria parasites in their salivary glands among those who survived the required incubation period (17 days).

3. The number of eggs.

4. The number of produced eggs.

5. The number of adult mosquitoes (F1)obtained from the eggs.

After feeding mosquitoes all animals were killed when the bleeding and serum samples were stored frozen (-20°C) prior to collection.

Any number of experimental compounds remaining in the end of the study, and kept frozen (-20°C) prior to collection.

Results

Graphical representation of the results of the experiments are shown in figa-3I and 4. In the first set of figures (figure 3) demonstrates the following data on the effect of the vaccine on fertility mosquitoes, for each of groups 1-5:

3A: Percentage of mosquitoes that received food.

3B: Average number of produced eggs.

3C: Average number of eggs.

3D: Percentage of hatching.

3E: the Average number of larvae.

3F: the Average number of pupae.

3G: the Percentage of pupation.

3H: Percentage departure.

3I: Average number of adult mosquitoes.

Numeric data supporting each of these graphical representations of a-I, placed respectively (column And DL is Figo etc) in table 2 below.

Table 2
Data for figures 3A-3I
Mean ± standard errorABCDE
Group 182,7±429,6±537,5±4,620,1±3,954,6±4,7
Group 266,7±6,517,1±3,812±39,7±2,451,9±7,4
Group 381,2±3,419,1±4,616±3,712,9±444,3±8,7
Group 473,7±4,924,1±3,926,4±3,915±2,856,6±6,6
Group 5to 75.7±4,530,1±4,132,4±516,9±2,5 55±4,6

Mean ± standard errorFGHI
Group 1the 15.6±3to 82.9±314±2,8to 92.4±2,6
Group 27,8±1,8of 86.1±5,76,9±1,880±7,8
Group 312,6±3,7to 77.7±7,312,5±3,692±1,8
Group 412,3±2,188,5±3,310,8±1,9and 88.8±2,4
Group 514,4±2,2to 88.4±2,712,1±1,8of 84.8±3,7

The data presented in tables 3A-3I below indicate the p-values obtained using non-parametric statistical analysis Mann-Whitney above data.

Table 3A
P-values for the percentage of mosquitoes that received food
Connection 1Connection 2Connection 3Connection 4
Group 1
Group 20,10504
Group 30,417270,14813
Group 40,148130,265430,10504
Group 50,232350,232350,265430,5

Table 3B
P-values for the average number of produced eggs
Connected to the e 1 Connection 2Connection 3Connection 4
Group 1
Group 24,6E-05
Group 30,000420,1612
Group 40,040380,00320,01602
Group 50,178360,00180,005510,22172

Table 3C
P-values for the average number eggs
Connection 1Connection 2Connection 3Connection 4
Group 1
Group 20,09451
Group 30,076260,49308
Group 40,273290,094160,10891
Group 50,306160,011710,008790,15145

Table 3D
P-values for the percentage hatching
Connection 1Connection 2Connection 3Connection 4
Group 1
Group 20,46767
Group 30,191550,29445
Group 40,366990,338030,14632
Group 50,400580,479070,150410,40187

Table 3E
P-values for the average number of larvae
Connection 1Connection 2Connection 3Connection 4
Group 1
Group 20,04897
Group 30,073710,45038/td>
Group 40,298010,061970,11695
Group 50,44990,01640,065240,23691

Table 3F
P-values for the average number of pupae
Connection 1Connection 2Connection 3Connection 4
Group 1
Group 20,06224
Group 30,286620,22088
Group 40,440570,04081,35186
Group 50,405460,011180,173730,25066

Table 3G
P-values for percentage pupation
Connection 1Connection 2Connection 3Connection 4
Group 1
Group 20,11277
Group 30,36640,13511
Group 40,118480,507980,07764
Group 50,106270,419710,08354,37388

Table 3H
P-values for percentage departure
Connection 1Connection 2Connection 3Connection 4
Group 1
Group 20,06876
Group 30,128150,25697
Group 40,0590,337120,2767
Group 50,020060,48530,185320,2943

Connection 1
Table 3I
P-values for the average number of adult mosquitoes
Connection 2Connection 3Connection 4
Group 1
Group 20,04391
Group 30,471270,06735
Group 40,39810,039080,47468
Group 50,464880,01230,313280,24603

Figure 4 shows the survival percentage of mosquitoes in each of the five groups during the time period of 8 days. The number of mosquitoes in each group at day 0 was as follows:

Group 1 (control):100
Group 2:89
Group 3:100
Group 4:99
Group 5:98

Data related to the graphical representation in figure 4, are presented in table 4 below.

Table 4
The data for figure 4
Mean ± standard errorDay 0Day 1Day 2Day 3Day 4
Group 1100±086±4,277±573±5,261±4,6
Group 2100±053,4±5,540,4±5,336,4±4,928±5,2
Group 3100±067±4,551±4,744±4,936±4,4
Group 478±5,162±5,760±5,651±4,9
Group 5100±073±5,653,4±6,748,4±6,341,4±5,6

Mean ± standard errorDay 5Day 6Day 7Day 8
Group 158±4,856±4,453±4,553±4,5
Group 223,7±521,4±4,820±4,820±4,8
Group 330±4,128±3,728±3,728±3,7
Group 446±546±543±5,243±5,2
Group 56,7±5 33,7±5,329,4±529,4±5

The data presented in table 5(i)-5(viii) below, show the p-values obtained using non-parametric statistical analysis Mann-Whitney above data.

Table 5(i)
P-values for day 1
Day 1Connection 1Connection 2Connection 3Connection 4
Group 1
Group 22,6E-05
Group 30,0021
7
0,0326
Group 40,132280,00070,0387
Group 50,0526 0,00710,15880,2738

Table 5(ii)
P-values for day 2
Day 2Connection 1Connection 2Connection 3Connection 4
Group 1
Group 21,1E-05
Group 30,00030,0676
Group 40,02740,004530,06759
Group 50,00490,078330,445980,166

Table 5(iii)
P-values for day 3
Day 3Connection 1Connection 2Connection 3Connection 4
Group 1
Group 21,1E-05
Group 3is 0.00020,20756
Group 40,049550,002530,0213
Group 50,003110,076920,267340,10538

Table 5(iv)
P-values for day 4
Day 4Connection 1Connection 2Connection 3Connection 4
Group 1
Group 23,8E-05
Group 30,000320,12615
Group 40,095150,001750,01525
Group 50,00650,047590,248540,09848

2,6E-05
Table 5(v)
P-values for day 5
Day 5Connection 1Connection 2Connection 3Connection 4
Group 1
Group 2
Group 36,9E-050,14515
Group 40,079760,001980,00944
Group 50,003930,039550,149620,09848

Table 5(vi)
P-values for day 6
Day 6Connection 1Connection 2Connection 3Connection 4
Group 1
Group 21E-05
Group 32,3E-050,12817
Group 40,120220,000810,00393
Group 50,00340,058020,221720,0558

Table 5(vii)
P-values for day 7
Day 7Connection 1Connection 2Connection 3Connection 4
Group 1
Group 22,3E-05
Group 38,4E-050,07147
Group 40,136480,001750,01315
Group 50,001320,090320,407950,03713

Table 5(viii)
P-values for day 8
Day 8Connection 1Connection 2Connection 3Connection 4
Group 1
Group 22,3E-05
Group 38,4E-050,07147
Group 40,136480,001750,01315
Group 50,001320,090320,407950,03713

EXPERIMENT 2: the Indus the Ktsia cytokine production by immunization which the polypeptide antigens

Peptides and recombinant proteins

The most effective in experiment 1 faction investigated further to identify polypeptides that can be used in vaccines. The effectiveness of the peptides of the present invention was determined using the following Protocol.

Immunization

All study polypeptides (drugs antigens) are synthesized by means of chemical technology, using Fmoc-protective group.

Of C57BL/6 mice aged six to ten weeks are subjected to immunization subcutaneous injection of doses of antigens in a volume of 200 μl per mouse. In the tested group each dose of the drug antigens contains equimolar mixture of peptides (10 nmol each), prepared in Freund (Sigma) according to manufacturer's instructions. In the control group each dose of the antigen has an equal dose of unrelated polypeptide prepared in IFA (Sigma) according to manufacturer's instructions (preparation NRP).

At day 15 after immunization, all animals receive a booster immunization using the same dose and route of administration, as well as the initial dose and route of administration.

Finally, at day 20 and all sort of animals and collect their spleen and serum.

ELISA of cytokines

Spleen of mice belonging to the same experimental group, unite, careful evaluation is to provide separation in a grid filter for cells and erythrocytes are removed by treatment with a buffer for lysis of erythrocytes (nine parts of 0.16 M NH 4Cl and one part of 0.17 M Tris, pH of 7.2). Suspension of splenocytes from each experimental group are seeded in 96-well plates in four repetitions at a density of 4 × 106cells/well in IMDM medium (Invitrogen)supplemented with 0.02 mm β-mercaptoethanol (Sigma), 50 IU/50 mg/ml penicillin/streptomycin (Sigma) and 10% FCS (Sigma) and containing each of the investigated antigen polypeptides (2 μm). After 3 days incubation at 37°C. the supernatant is collected and analyzed for IFN-γ and IL-4 using a sandwich ELISA for cytokines in accordance with the manufacturer's Protocol (Pharmingen). The lower limit of detection for this assay are 9,77 PG/ml for IL-4 and 39,06 PG/ml for IFN-γ.

Specific IgG2a ELISA

96-well plates to micrometrology used for ELISA (Becton-Dickinson), cover with 2 mm of each experimental polypeptide in PBS. After incubation over night at 4°C tablets washed twice in PBST (PBS containing 0.5% Tween-20) and the wells blocked with 1% fraction V BSA in PBST. After incubation for 1 h tablets washed three times in PBST and holes add a series of dilutions of the tested and control sera in PBST. After incubation for 2 h tablets washed six times in PBST and to all wells add primary serum against Ig2a mouse. After incubation for 1 h tablets washed six times in PBST and to all wells add serum against primary sera against Ig2a Michiel incubation for 1 h tablets washed seven times in PBST and to all wells add the TMB substrate. After incubation for 20-30 minutes and the reaction stopped by the addition of HCl and read optical density at 450 nm.

Statistical analysis

Statistically significant differences in the production of IFN-γ in response to various antigens between the test and control groups set due to the non-parametric Mann Whitney analysis of samples. Differences are considered statistically significant if the p-value is below 0.05.

EXPERIMENT 3 - Evaluation of immune response to various polypeptides

Were studied the following polypeptides: SEQ ID NO:20, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32 and SEQ ID NO:35. These peptides were mixed together with the formation of a vaccine candidate to test (called AGS-peptides).

The type and extent of the immune response induced by immunization with these peptides was evaluated according to the following Protocol:

Day 1: Give 2 groups of 4 mice CD1 following each injected subcutaneously with doses of product candidates for the vaccine:

- A mixture of irrelevant peptides (NRP) (10 nmol each) + ISA-51

- AGS-mixtures of peptides (10 nmol each) + ISA-51

Day 15: to Carry out the booster immunization all animals the same doses of product candidates for the vaccine.

Day 21: In the conclusion to bleed all the animals. Collect separately spleen and check for the products of IFN-gamma in response to:

Separate the peptide AGS (2 μm each)

- AGS-mixture (0.5 µm and 2 µm of each)

- Concanavalin

Control

After day 21: to Test all serum on the ability to react with peptides AGS.

Results

Production of IFN-gamma after 96 hours of stimulation in vitro antigens shown in the diagram in figure 5.

Production of IFN-gamma in response to SEQ ID NO:28, SEQ ID NO:30 and SEQ ID NO:35, and the drug AGS-mixtures are statistically significant (p<0,05).

SEQ ID NO:20, SEQ ID NO:31 and SEQ ID NO:32 induce a response greater extent in animals immunized with AGS-mix, but they also, apparently, are stimulating, non-specific way, splenocytes from mice immunized with NRP-mixture.

Education Ig in General in the serum in response to the antigens shown in the diagram of figure 6. Education Ig as a whole in response to SEQ ID NO:20 and SEQ ID NO:30 are statistically significant (p<0,05).

EXPERIMENT 4 - a Study of infection after immunization AGS-peptides

To test the ability of the drug AGS mixture to provide protection from natural malaria infection CD1 mice were subjected to immunization and infection in accordance with the following Protocol:

Day 0: Take blood samples from all animals. Save the samples for further analysis.

Day 1: Give 2 (two) groups of 8 mice CD1 each following subcutaneous injection of doses of product candidates for the vaccine:

- With whom thou irrelevant peptides (NRP) (10 nmol each) + ISA-51

- AGS-mixtures of peptides (10 nmol each) + ISA-51

Day 7: Take blood samples from all animals. Save the samples for further analysis.

Day 14: to Make the booster immunization of animals with the same doses of product candidates for the vaccine.

Day 21: Take blood samples from all animals. Save the samples for further analysis.

Day 28: infect All animals through bites of 8 infected mosquitoes (i.e. Anopheles gambiae infected with Plasmodium yoelii nigeriensis) in the abdominal region. All animals remain to first establish the presence of parasites or for a maximum of 6 weeks after infection by infected mosquitoes.

Day 70 (maximum): In conclusion, to bleed all animals.

Results

Education Ig in General in the serum in response to the AGS mixture is shown in graph 7.

One animal in the group immunized with the AGS mixture demonstrated a significantly lower education Ig in General than the rest of the animals in this group (<50% secondary education Ig in General in the group immunized with AGS-mix).

On the day of infection one animal in the group immunized with NRP-mixture, and two animals in the group immunized with the AGS mixture, it was impossible to infect due to the lack of infected mosquitoes.

Among the exposed to infected animals animals in the group immunized with the AGS mixture showed Bo the greater degree of survival, than animals in the control group immunized with NRP-mixture (see Fig). One animal that died in immunized AGS-mixture group was the same animal, which has developed a strong humoral immune response to the drug AGS.

1. Polypeptide composition comprising one or more polypeptides that are immunogenic in the spinal, so they cause spinal products of cells of the immune system that can recognize at least one epitope in the protein fraction of arthropod saliva, and protein fractions of arthropod saliva has mass, component <20 kDa, and polypeptides independently selected from:
the polypeptide sequences SEQ ID NO:1-28, 33-39, 44 or subsequences of these sequences having 7 amino acids or more:
SEQ ID NO:1 HLTLFTVAVLLLAAAALLLLLPPAYSTTLTPP
SEQ ID NO:2 PLSYCHLFLTHTLARALSFSRSDCL
SEQ ID NO:3 KNVFFALLLWLVCCLVSVQGNEI
SEQ ID NO:4 KLLVLLICLFFYHTHCTTAYLWLAMGV
SEQ ID NO:5 FLKGSFPRFQMCVMLIGFFSSAKCL
SEQ ID NO:6 NDYQALLGLCCPWIDLAAADLPMRRHAKA
SEQ ID NO:7 FYSVGKLVKVLLVMAVCCLLLCTAPTGADPL
SEQ ID NO:8 MKFAFAFVLIALFAVFAVSQALPQPEQAAA
SEQ ID NO:9 DGASAITKIVLELTPEQAAAV
SEQ ID NO:10 TLFIFLVCCQIPLFGIMSSDSADPFYWIRVILA
SEQ ID NO:11 GRVMCLLRLMSTLLWLSIVGK
SEQ ID NO:12 LYSGYRLLVLLVMTVCCLLLFIAPTGADPLPGQTQRTL
SEQ ID NO:13 MYCVIKGKTGGYCNSEGLCTCRAEDLHFLLKPIINKD
SEQ ID NO:14 NAEDPRTELIGCGSVLFHLAANRLSLQLEEFAVCKR
SEQ ID NO:15 ALIGLLLCSVQSVTANDPVDALGACSGNLFGLLMTRL
SEQ ID NO:16 SKLFVLAFLCLALWWQSAPQYARGDVPT
SEQ ID NO:17 SMLVAFATLSVALWWAIPANFNYGGGGGYFINGTGQ
SEQ ID NO:18 IYEKLPAYLSEVSARVNVLQVSLQHDLPNLQ
SEQ ID NO:19 EMKLAKVALVTISLWFMAWTPYLVINFTGI
SEQ ID NO:20 LLPAKVIPDKTAAYVAYGGQTLVEHVEVLV
SEQ ID NO:21 FYTCFLGTSSLAGFKNAVDYDELLKAG
SEQ ID NO:22 VLEVLGFVEDNGELVFQELLGVLKMVDPDGD
SEQ ID NO:23 KLTPTWWLLCLTFVADALTIQELRAQIAQQRIQQRYGVTVATT
SEQ ID NO:24 SLSDYGLIELKEHCLECCQKDTEADSKLKVYPAAVLEV
SEQ ID NO:25 TYICFILHGVSEIIPQQQKKTMKFLLLVASVLCLVLI
SEQ ID NO:26 RYFWIALICPLIIVETLAV
SEQ ID NO:27 LLLYLDAADLRRALHQYQLLAAQGDRHLPQQIVKFV
SEQ ID NO:28 VLLTPALQAYIMDEHNLNRSNIALGRIRPYPSAVKMP
SEQ ID NO:33 KFYRLISTLLWWIAPRHQCSPFFFQYNRPYL
SEQ ID NO:34 NYVPDVSALEQDIIEVDPETKEMLKHLDFNNIWQL
SEQ ID NO:35 QYSMECLEAAEPKYLDGLKTLADETAQC
SEQ ID NO:36 EYAQVTKMLGNGRLEAMCFDGVKRLCHIRGKL
SEQ ID NO:37 KLFLTLLSTLSVAMVFALPAHHHSRG
SEQ ID NO:38 ELEEARLVAEELEERQQELDYLKRYLVGRLQAV
SEQ ID NO:39 SYFLTVCLLALVQSETVQD
SEQ ID NO:44 LLLAMVLLPLLLLESWPYAAAEKVW.

2. Polypeptide composition according to claim 1, where the composition
optionally contains one or more sequences independently selected from:
the polypeptide sequences SEQ ID nos:29-32 or 40-43 or subsequences of these sequences having 7 amino acids or more:
SEQ ID NO:29 VLKGETHKALKLKDGGHYLVEFKSIYM
SEQ ID NO:30 VLHSMLVNASLAEMVKESYQTHGADGRMWRMLKFVRLLP
SEQ ID NO:31 RVRALRALLETLLQHQGEQNNDVYLIRLAHET
SEQ ID NO:32 ELQQALSSLNAGSGSCAEVFNAYLPVHNKYIGVSRKI
SEQ ID NO:40 AMTNANLVGLTISLAYAIFFLLYTPPTGRSS
SEQ ID NO:41 SFAWLLYGIILRSNFLWQNLMALALSAVQLSLFII
SEQ ID NO:42 AFPFISGFLSCFMWLKYGVLTEESTLILVNFIGSAL
SEQ ID NO:43 GLLCCCLAVLFFASPLTMLAHVIR.

3. Polypeptide composition according to claim 1 or 2, which contains 2-12 polypeptides or more preferably 2-6 polypeptides.

4. Polypeptide composition according to claim 1, in which one or more polypeptides are epitopes of cytotoxic T lymphocytes (CTL)and/or one or more polypeptide epitopes are T-cell-helper cells (Th), and/or one or more what about the polypeptides are epitopes of b-lymphocytes.

5. Polypeptide composition according to claim 1, containing 2, 3, 4, 5, 6 or more epitopes as defined in claim 1.

6. Polypeptide composition according to claim 1, which is immunogenic in respect of multiple proteins of arthropod saliva.

7. Polypeptide composition according to claim 1, in which the protein fraction of arthropod saliva is the protein fraction of the saliva of any mosquito species, preferably Anopheles gambiae.

8. Polypeptide composition according to claim 1, additionally containing one or more additional sequences of proteins of arthropod saliva.

9. Polypeptide composition according to claim 1, in which the subsequence contains 8, 9, 10 or 11 amino acids or more.

10. Polypeptide composition according to claim 1, essentially consisting of two or more polypeptides having the sequence of sequence SEQ ID NO:1-28, 33-39, 44.

11. Polypeptide composition according to claim 1, in which there are all polypeptides having the sequence of SEQ ID NO:1-6, and/or which contains all of the polypeptides having the sequence of SEQ ID NO:20, 28, 30-32 and 35, and/or which contains all of the polypeptides having the sequence of SEQ ID NO:6, 9, 29, 31 and 35, and/or which contains all of the polypeptides having the sequence of SEQ ID NO:6, 9 and 35.

12. Polypeptide composition according to claim 1, used in medicine.

13. Polypeptide composition according to claim 1, usage is suuwassea in veterinary medicine.

14. Polypeptide composition, which contains a polypeptide composition according to any one of claims 1 to 13, and the media.

15. Polypeptide composition according to 14, in which the carrier includes an adjuvant and/or excipient.

16. A method of obtaining a polypeptide composition according to any one of claims 1 to 13, comprising mixing or combining one or more polypeptides as defined in any one of claims 1 to 13, with one or more polypeptides as defined in any one of claims 1 to 13, and/or one or more additional components, such as carrier, excipient, adjuvant, buffer or stabilizer.

17. A method of obtaining a polypeptide composition according to item 16, which includes linking, combining or mixing the polypeptide composition according to any one of claims 1 to 13 with the carrier.

18. The method according to 17, in which the carrier includes an adjuvant and/or excipient.

19. Drug against distributed arthropod disease containing polypeptide composition according to any one of claims 1 to 13 and, optionally, a suitable excipient and/or adjuvant.

20. Drug in claim 19, where the arthropods is a blood-sucking claystone.

21. Drug in claim 20, where the blood-sucking claystone choose from mosquitoes, blood-sucking ticks, sand mosquitoes, ticks subclass Acarina and fleas.

22. Medical environments, the creation in claim 20, where claystone choose from Anopheles sp., Aedes sp., Culex sp., Mansonia sp., Chrysops sp., Simulium sp., Tnatoma sp., Rhodnius sp., Panstrongylus sp., Glossina sp., Tabanus sp., Phlebotomus sp., Lutzomyia sp., Pediculus sp., Ornithodoros sp., Ixodes sp., Pediculus sp., Pulex sp., Hemagogous spegazzani, Ochlerotatus sp. and Coquillettidia sp.

23. The drug according to any one of p-22, where distributed arthropod disease is a disease selected from malaria, filariasis, trypanosomiasis, leishmaniasis, onchocerciasis, dengue, disease Lima, tularemia, relapsing typhus, epidemic typhus, typhoid, plague, yellow fever, rift valley fever and encephalitis, including encephalitis St. Louis encephalitis West Nile, Eastern equine encephalitis, encephalitis, and La Crosse and Eastern equine encephalitis.

24. The method of obtaining drugs in claim 19, which includes linking, merging or blending of the composition of the nucleic acid according to any one of claims 1 to 13 with a suitable excipient and/or adjuvant.

25. The composition of the vaccine distributed arthropod disease containing composition according to any one of claims 1 to 13 and, optionally, a suitable excipient and/or adjuvant.

26. The composition of the vaccine on A.25, where the arthropods is a blood-sucking claystone.

27. The composition of the vaccine for p, where blood-sucking claystone choose from mosquitoes, blood-sucking ticks, sand mosquitoes, klimapolitik Acarina and fleas.

28. The composition of the vaccine for p where claystone choose from Anopheles sp., Aedes sp., Culex sp., Mansonia sp., Chrysops sp., Simulium sp., Triatoma sp., Rhodnius sp., Panstrongylus sp., Glossina sp., Tabanus sp., Phlebotomus sp., Lutzomyia sp., Pediculus sp., Ornithodoros sp., Ixodes sp., Pediculus sp., Pulex sp., Hemagogous spegazzani, Ochlerotatus sp. and Coquillettidia sp.

29. The vaccine composition according to any one of PP-28, where distributed arthropod disease is a disease selected from malaria, filariasis, trypanosomiasis, leishmaniasis, onchocerciasis, dengue, Lyme disease, tularemia, relapsing typhus, epidemic typhus, typhoid, plague, yellow fever, rift valley fever and encephalitis, including encephalitis St. Louis encephalitis West Nile, Eastern equine encephalitis, encephalitis, and La Crosse and Eastern equine encephalitis.

30. A method of obtaining a vaccine composition according A.25, which includes linking, merging or blending of the composition of the nucleic acid according to any one of claims 1 to 13 with a suitable excipient and/or adjuvant.

31. The method of treatment or prophylaxis of disseminated arthropod disease involving the introduction of a vertebral polypeptide composition according to any one of claims 1 to 15, the medicinal product according to any one of p-23 or vaccine according to any one of p-29.

32. The method according to p, in which the vertebrate is selected from a mammal, bird, reptile and fish, and preferably from human, Domash is it an animal or a farm animal, cattle, or poultry.

33. The method according to p or 32, where the arthropods is a blood-sucking claystone.

34. The method according to p, where blood-sucking claystone choose from mosquitoes, blood-sucking ticks, sand mosquitoes, ticks subclass Acarina and fleas.

35. The method according to p where claystone choose from Anopheles sp., Aedes sp., Culex sp., Mansonia sp., Chrysops sp., Simulium sp., Triatoma sp., Rhodnius sp., Panstrongylus sp., Glossina sp., Tabanus sp., Phlebotomus sp., Lutzomyia sp., Pediculus sp., Omithodoros sp., Ixodes sp., Pediculus sp., Pulex sp., Hemagogous spegazzani, Ochlrtatus sp. and Coquillettidia sp.

36. The method according to any of PP-35, where distributed arthropod disease is a disease selected from malaria, filariasis, trypanosomiasis, leishmaniasis, onchocerciasis, dengue, Lyme disease, tularemia, relapsing typhus, epidemic typhus, typhoid, plague, yellow fever, rift valley fever and encephalitis, including encephalitis St. Louis encephalitis West Nile, Eastern equine encephalitis, encephalitis, and La Crosse and Eastern equine encephalitis.

37. The use of a polypeptide composition according to any one of claims 1 to 15 for the manufacture of a medicinal product or a vaccine that is effective for the treatment or prophylaxis of disseminated arthropod disease.

38. The application of clause 37, where the arthropods is a blood-sucking claystone.

39. The application of § 38, where the blood-sucking claystones is selected from mosquitoes, blood-sucking ticks, sand mosquitoes, ticks subclass Acarina and fleas.

40. The application of § 38, where claystone choose from Anopheles sp., Aedes sp., Culex sp., Mansonia sp., Chrysops sp., Simulium sp., Triatoma sp., Rhodnius sp., Panstrongylus sp., Glossina sp., Tabanus sp., Phlebotomus sp., Lutzomyla sp., Pediculus sp., Ornithodoros sp., Ixodes sp., Pediculus sp., Pulex sp., Hemagogous spegazzani, Ochlerotatus sp. and Coquillettidia sp.

41. Use on any p-40, where distributed arthropod disease is a disease selected from malaria, filariasis, trypanosomiasis, leishmaniasis, onchocerciasis, dengue, Lyme disease, tularemia, relapsing typhus, epidemic typhus, typhoid, plague, yellow fever, rift valley fever and encephalitis, including encephalitis St. Louis encephalitis West Nile, Eastern equine encephalitis, encephalitis, and La Crosse and Eastern equine encephalitis.

42. The composition of nucleic acids containing one or more nucleic acids comprising a sequence encoding one or more polypeptides in the polypeptide composition according to any one of claims 1 to 13.

43. The composition of the nucleic acid according to § 42, which is composed of DNA or RNA.

44. The composition of the nucleic acid according to § 42 or 43, which contains a plasmid or a recombinant virus.



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to abdominal surgery and oncology, and may be used for treating patients with tumour intestinal obstruction. The method involves gastrointestinal decompression, active lavage, enterosorption, enteral nutrition. The enteral therapy is intraoperative. The minimum early enteral nutrition is prescribed on the first postoperative day and thereafter; adequate tolerance of the probe nutrition and the presence of minimum excretion allows alternation of various formulae for enteral nutrition. On the first postoperative day, a monomer formula with added 20% Nutriene Elemental 200 ml is introduced. On the second day, Nutriene Elemental is introduced in the volume of 10 kcal/day. On the third day, Nutrisone Standard is introduced in the volume of 15 kcal/day. On the forth day, the formula Nutrisone Energia with dietary fibres is introduced in the volume of 20 kcal/day. On the fifth day, after the nasointestinal probe has been removed and unassisted stool is observed, Pevzner's therapeutic diet No.1 which is added by the introduction of the formula Nutrisone Energia with dietary fibres by sipping method.

EFFECT: method provides clinical effectiveness including ensured by correction of protein-calorie and intestinal insufficiency in the patients with acute intestinal obstruction of tumour aetiology.

1 tbl, 2 ex

FIELD: food industry.

SUBSTANCE: invention is related to food industry and may be used for food allergy prevention. Hydrolysed by fermentative method egg whites with hydrolysis degree from 15% to 28% are applied in production of food compositions.

EFFECT: invention ensures tolerance to egg whites contained in food products with mammals.

5 cl, 5 dwg, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of pharmaceutics. Composition contains botulinum toxin, polysorbate 20 and methionine, where methionine is present in amount from 25 to 75 mM, and polysorbate 20 is present in amount from 0.1 to 2.5 mg/ml.

EFFECT: composition possesses improved stability; application of liquid pharmaceutical composition in form of combination of polysorbate 20 and methionine as botulinum toxin stabilisers instead of proteins of animal origin albumin and gelatin eliminates risk of organism contamination by pathogens or microorganisms of serum origin and makes it possible to introduce into organism safely; in addition, composition is convenient for application in direct introduction to patients.

8 cl, 8 tbl

FIELD: medicine.

SUBSTANCE: group of inventions relates to medicine, namely to resuscitation science and intensive therapy, and can be used for providing feeding of critically ill patients or patients exposed to stress. For this purpose first nourishing preparation with specified composition is introduced to patient in SIRS phase, and in phase CARS second nourishing preparation of specified composition is introduced.

EFFECT: application of claimed inventions reduces risk of infection and body weight loss increasing probability of recovery due to reduction of oxidative stress and muscle decomposition.

FIELD: medicine.

SUBSTANCE: invention represents application of composition, which includes protein material, where said protein material provides, at least, 24% of energy value of composition (en.%) and, at least, 12 wt % of all protein material is represented by leucine, for preparation of medication for treatment of subject with obesity.

EFFECT: extension of assortment of compositions, efficient for treatment of obesity.

35 cl

FIELD: medicine.

SUBSTANCE: invention refers to pharmaceutical industry, particularly to ointment for treating burn diseases. The ointment for treating burn diseases, containing as an active substance, dry albumin and ethanolic propolis extract, and as an ointment base, salve base, penthol and purified water in a certain ratio of components.

EFFECT: ointment exhibits reparative, antiinflammatory, dehydrating, necrolitic and antimicrobic action and is effective for treating burn diseases.

5 ex

FIELD: medicine; veterinary science.

SUBSTANCE: method provides inoculation and incubation of spore culture Bacillus subtilis 3 in a nutrient medium containing placental embryonic utricular hydrolyzate of animals' tissues and organs containing 90 mg % amine nitrogen and prepared from placenta, embryos and uterus of butchers to produce a culture fluid of culture Bacillus subtilis 3. Then montmorillonite fraction of zeolite is produced by conditioning in hydrochloric acid followed by removal of soluble salts and quartz with distilled water. There are remained zeolite particles of size 0.0006-0.0009 mm. Then the fraction is dried. After that, suspending montmorillonite fraction of zeolite and milk albumin hydrolyzate are added to the culture fluid Bacillus subtilis 3. The culture fluid Bacillus subtilis 3, suspending fraction of zeolite, and milk albumin hydrolyzate are mixed in the ratio 1:1:0.5 respectively. The produced preparation is sterilised and introduced once in doses 7-10 cm3 (to small animals) and 15-20 cm3 (to large animals) at 6.5-7.5 mg of solid per 1 kg of live weight within first postradiation 1-10 days.

EFFECT: higher protective effectiveness from radiation injuries and reduced application time.

3 cl, 1 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: there is offered application of the substance representing enzyme-processed fish protein hydrolyzate able to inhibit activity of acyl-CoA cholesterole acyltransferase and to intensify mitochondrial β-oxidation for making a pharmaceutical or food composition applied for treatment and/or prevention of hepatic adipose degeneration in animals, for treatment and/or prevention of hypercholesterolemia and for treatment and/or prevention of hyperhomocysteinemia, and also for treatment and/or prevention of atherosclerosis, coronary heart disease, stenosis, thrombosis, myocardial infarction and stroke in an animal. There is offered method for making enzyme-processed fish protein hydrolyzate able to inhibit activity of acyl-CoA cholesterole acyltransferase, to reduce triglyceride concentration in liver, to reduce homocysteine concentration in plasma and/or to intensify mitochondrial β-oxidation. The preferential embodiment of the invention concerns FPH application as an antiatherogenic and cardioprotective agent presented either as a pharmaceutical agent, or as a functional food.

EFFECT: improved properties of the substance.

14 cl, 7 ex, 5 tbl, 4 dwg

FIELD: medicine.

SUBSTANCE: invention is related to medicine, namely, to orthopaedic stomatology and may be used in treatment of inflammatory diseases of mouth tunica mucosa in area of dentures and/or implants. For this purpose affected section of tunica mucosa is coated with argacol of 0.3-0.6 mm layer. Treatment is carried out 2-3 times per day for 5-14 days.

EFFECT: application of argacol in specified mode makes it possible to provide for high treatment effect with no unpleasant feelings of patient.

2 cl, 2 ex

FIELD: food products.

SUBSTANCE: invention relates to compositions and food products made of oat fractions. Compositions includes at least 25 wt % of protein and at least 1 wt % of emulsifying lipids as water-insoluble mixture and not more than 8 wt % of β-yeast cellulose on dry weight basis count. Protein and emulsifying lipids are obtained of the same oat fraction. Mass ratio protein and emulsifying lipids makes from 1: 0.01 to 1: 0.6. Compositions includes at least 15 wt % of protein hydrolizate and at least 1 wt % of emulsifying lipids as water-insoluble mixture. Protein and emulsifying lipids are obtained of the same oat fraction which is hydrolised. Mass ratio of protein hydrolizate and lipids is from 1:0.01 to 1:1. Food product includes at least one main nutritious element and composition. Also method of obtaining mentioned compositions is shown.

EFFECT: improvement of wheyey lipidic profile.

2 tbl, 16 ex

FIELD: biotechnology, biochemistry.

SUBSTANCE: invention relates to producing the biologically active complex eliciting antioxidant and immunomodulating activity and used in medicine, cosmetics, veterinary science and food industry. The biologically active complex preparing by enzymatic hydrolysis of muscle tissue represents complex of biologically active compounds involving carnosine and anserine in the amount 85-97 wt.-% of the native content of these components in poultry muscle tissue, 1-7 weight parts of amino acids, 0.5-12 weight parts of oligopeptides of molecular mass 10 kDa, not above, and 0.1-15 weight parts of cyclic and polycyclic phenolic compounds as measured for 1 weight part of carnosine and anserine in the complex. This complex is prepared by enzymatic hydrolysis of milled and homogenized water muscle tissue in preferable dilution homogenate with water in the range 0.2-0.6 and with using proteolytic enzymes in the amount 2-5 wt.-% of the protein content and working at pH 4.5-8.5 and at enhanced temperature being preferably at 45-65°C. Product is isolated as extract or powder prepared in drying the extract. Invention provides enhancing effectiveness of the claimed complex.

EFFECT: improved method for preparing, valuable properties of complex.

7 cl, 6 tbl, 6 ex

FIELD: medicine.

SUBSTANCE: the present innovation deals with treating vestibular and auditory disorders, trigeminal nerve's neuralgia and peripheral paresis of facial nerve of herpetic etiology. The method deals with introduction of an antiherpetic preparation followed by immunomodelling therapy with the use of polyoxidonium. Moreover, after treating with antiherpetic preparation before introducing polyoxidonium it is necessary to conduct additional successive therapy: with preparations of trophic action as milgamma or neuromultivit at simultaneous introduction of antioxidant, then comes intratissue injection of cerebrolysine being behind the top of mastoid process.

EFFECT: the innovation enables to decrease the quantity of relapses due to stopping the development of herpes simplex virus, restore conductivity along nervous fiber and improve endoneural circulation.

3 cl, 3 ex

FIELD: medicine.

SUBSTANCE: method involves applying antiherpetic therapy using Phamcyclovir. Then, Trental is sequentially introduced during 5 days with 2-5 ml of cerebrolysin being concurrently intratissularly introduced behind mastoid process tip. Mexidol is administered for 10 days. The treatment is finished by introducing Polyoxydonium.

EFFECT: combined microcirculation improvement; high neurotrophic and antioxidant activity; secondary immune deficiency adjustment; eliminated labyrinth hydrops.

FIELD: medicine, neurology, in particular treatment of disseminated sclerosis.

SUBSTANCE: complex therapy includes plasma exchange, interferonotherapy, administration of copaxon, cytostatics, symptomatic and bracing agents, and cyclosporin A. Administration of steroids is excluded. Additionally ceruloplasmine intravenously drop-by-drop in dose of 100 mg and cerebrolysate intramuscularly in dose of 10 ml for 10 days are administered. Claimed method provides stable remission up to 12 months for 89.2 % of patients.

EFFECT: decreased invalidisation due to reconstitution of regulatory relationship between nervous and immune systems.

1 ex, 1 tbl

FIELD: medicine, neurology, pediatrics.

SUBSTANCE: method involves administration of aminolone in the dose 0.25-0.5 g, 2-3 times per a day for 6-8 months, cerebrolysine in the dose 1.0-1.2 ml by intramuscular route, course 15-20 injections, mydocalm in the dose 50-75 mg, 2 times per a day for 1-1.5 month, vitamin B12 in the dose 150-300 mcg, course 15-20 injections wherein injections of vitamin B12 and cerebrolysine are alternated in each other day; in 1.5 month after onset of this treatment prefizone in the dose 1 ml by intramuscular route is administrated every day, course 15-20 injections, pyrogenal in the dose 100-200 MTD is administrated by course 15-20 injections and curative physical culture is carried out with heat procedure in sauna (dry bath) for 3 months, and after termination of medicinal therapy hyppotherapy is carried out for 3-5 months for each other day. Method allows straightening the kyphoscoliotic carriage and to improve joins mobility due to increase of the motions volume in them. Invention can be used in treatment of spastic diplegia in infantile cerebral paralysis.

EFFECT: improved treatment method.

3 ex

FIELD: medicine, ophthalmology.

SUBSTANCE: after termination of surgery operation as to strabismus the method involves catheterization of Tenon's space by using the irrigative system for the successive fractional administration of preparations of riboflavin, cerebrolysin, dicynone, caffeine, emoxipine and taufon in the dose 0.3 ml for each drug. Also, method involves carrying out the computer training by designation of the complex "Shooting range/gallery" and "Pursuit" wherein the patient combines color symbols that change colors and sizes in cyclic change of positions, 1 séance for each type of exercise every day. Administration of drugs and computer trainings are carried out for 10 days. Such performance of the method provides the effective treatment of strabismus and recovery of binocular vision based on the complex effect on all links of coordination activity of sensor and motor eye apparatus. Invention can be used in treatment of strabismus.

EFFECT: improved and effective method of treatment.

2 ex

FIELD: veterinary, in particular agents for prophylaxis and treatment of gastric diseases in calves.

SUBSTANCE: the fist foremilk of calved in summer cows is collected, filtered, bottled in sterile 1 l vessels and frozen in freezing apparatus at 20-22°C. In winter-spring period foremilk is defrosted and conserved with potassium sorbate in ratio of 2.5 ml of 40 % potassium sorbate solution per 1 l of foremilk.

EFFECT: environment friendly method for stimulation of calves immunity.

3 tbl

FIELD: medicine, biochemistry, pharmacy, biotechnology.

SUBSTANCE: invention relates to a method for preparing polyelectrolyte microparticles containing the end substance and showing sensitivity to alteration of the environment composition. Method involves preparing oppositely charged polyelectrolytes on microaggregates containing an encapsulated substance. These polyelectrolyte microparticles can be used both in medicine as systems used in delivery drugs and providing pH-sensitive release of encapsulated substance and in biotechnology as biocatalysts stabilized with respect to unfavorable conditions. Invention provides preparing polyelectrolyte microparticles characterizing by the high content of active substance - up to 90% of microparticles mass. Proposed method is sample and involves lesser amounts of steps.

EFFECT: improved preparing method.

9 cl, 3 tbl, 2 dwg, 61 ex

FIELD: medicine.

SUBSTANCE: method involves applying mono therapy with Cerebrolysin intramuscularly introduced once a day in the morning at a daily dose of 0.1 ml/kg of weight as 20 days long course or Encephabol to be taken 3 times a day in 30 min after having taken meals. Encephabol is given at a dose of 50 mg in 2-5 years old children cases or at a dose of 100 mg in 6-10 years old children cases. Male children take Encephabol in not longer than 10 days long course. Female children take Encephabol in a course being longer than 10 days.

EFFECT: enhanced effectiveness of genome instability adjustment.

6 dwg

FIELD: medicine.

SUBSTANCE: method involves applying nutritive probe support with 1-2% Pecto solution in Rehydron at a dose of 30.0-50.0 ml per one feeding session 6-7 times a day beginning in 24-48-h from receiving injury. Daily quantity of introduced solution is gradually increased to 500.0 ml. Then, semi-element food mixtures are introduced in 20% dilution from a standard one at a dose of 30.0-60.0 ml per one feeding session with 2.0-2.5 h long pause 7 times a day. Mixture dilution is gradually increased to standard one and volume to 1500-2000 ml. Pecto introduction is continued as solution of 5.5 g per 50.0 ml of water at a dose of 5.0-20.0 ml per 20 min before every probe-mediated feeding.

EFFECT: enhanced effectiveness in supporting energetic and plastic organism needs; avoided stress ulcer formation.

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