The way the specific prevention of aleutian mink disease

 

The invention relates to biology, medicine, immunology, biotechnology, immunopharmacology, fur farming. The aim of the invention is to obtain vaccine drug for the prevention of acquired immunodeficiency virus - Aleutian mink disease. This goal is achieved through the use as a vaccine preparation antiidiotypic antibodies or F(ab) fragments, which immunochemical analogues ("mimic") of the main antigens of viruses Aleutian mink disease. The technical result is to reduce morbidity and mortality mink 3-4 times. table 1.

The technical field: biology, medicine, biotechnology, immunology, immunopharmacology, farming TECHNIQUES LEVEL Aleutian disease of mink (ABN) is a species-specific acquired immunodeficiency virus. To a certain extent ABN homologous syndrome acquired immunodeficiency virus human. ABN is a highly contagious disease spread primarily with food and water and almost completely damaging the stud holes when the infection in the fur farms. ABN leads to a sharp reduction in fertility and a significant increase in mortality of the animals associated in frequent is Eesa and with the spread of the disease are struggling with the destruction of infected animals and the burning of used equipment.

Accordingly, the feasibility of implementation in fur practice effective way to prevent ABN obvious and does not require special justification.

In addition, taking into account the pronounced similarity of the acquired immunodeficiency virus - ABN mink and AIDS human development and testing of effective prophylactic vaccines against ABN can be an important step in obtaining preventive vaccine, "Anti-AIDS".

It is important that modern theoretical immunology and immunobiotechnology create preconditions for the development of simple and effective ways to get vaccines, based on the substitution of viral antigens (which are usually expensive, difficult, time consuming, and in some cases dangerous to health) they are cheaper and safer counterparts. As such analogues can theoretically be used specific antiidiotypic antibodies (AIAT) or Fab fragments (Fb2-fragments), because of "molecular mimicry" which immunochemical analogs of viral antigens.

The basis for this patent on the results of experiments in which he tested the possibility that AYAT or Fab-fragments of AIAT received on antiviru the vaccine, suitable for prevention of ABN.

Analogues of the described method is not available.

As a prototype of the present invention can be considered theoretical work describing the potential use of AIAT corresponding specificity for the treatment of bacterial and viral infections (Kingman, New Sci., 1989, 121, 1650, 30; Monroe a. Greene, Immunol. Investigations, 1986, 15, 3, 263-286). Note that this theoretical work has so far not gone beyond research laboratories.

DISCLOSURE OF THE INVENTION 1. Obtaining amounts of viral antigens that are targets of antibodies in animals, sick ABN.

Conventional methods (for example, by using a combination of ammonium sulfate precipitation and ion exchange chromatography on DEAE-cellulose) get the total fraction of immunoglobulins from serum mink, patients ABN and healthy mink.

Prepare extracts of kidney patients and healthy animals, homogenizing tissue in extracting water-salt buffer (e.g., on the basis of 0.05 M phosphate buffer, pH 7.5, containing 0.3 M NaCl and 0.05% Triton X-100), centrifuged for clarification and filtered to eliminate particles.

Pairs (separately) mixing the extracts of kidneys healthy mink and patients mink solutions selected complexes (for example, the polyethylene glycol-6000) and spend the incubation of the resulting mixtures (10-24 hours at+2...+8oC).

Centrifuged the mixture to obtain a precipitate of the precipitates. Washed precipitates extracting buffer, dissolve them and carry out electrophoretic separation of material precipitates in the polyacrylamide gel according to standard methods.

Analysis of electrophoregram of precipitates obtained with the use of immunoglobulins and extracts of kidney patients and healthy mink, identify and cut out from the gel zone, found only in the precipitates kidney mink, patients ABN and deposited only immunoglobulins (antibodies), isolated from the blood of sick animals. In this way we obtain trace amounts of "major" virus antigens ABN, which induces the production of the main anti-virus antibodies by the immune system of infected animals.

2. Receiving antiviral antibodies from the blood of mink, patients ABN.

Viral antigens identified in electrophoregram of immunoprecipitated, as described above, were extracted from the gels and immobilizovana on inert carriers for affinity chromatography (e.g., CNBr-sepharose, according to the instructions of the manufacturer Pharmacia, Sweden).

Obtained is passed through them in 2-5 ml of blood serum mink patients ABN. After washing the columns from unbound material, the elution of antiviral antibodies, contacting the immobilized viral antigens (e.g., 3 m solution of NaSCN), and conducted their dialysis against any neutral buffer solution.

To eliminate possible impurity materials obtained antibody was passed through a column with immobilized antigens extracts of kidneys healthy mink.

Purified anti-virus antibodies were subjected to hydrolysis by conventional methods to obtain F(AB) fragments or F(AB)2-fragments.

3. Obtaining specific antiidiotypic antibodies, used as a vaccine preparation.

Derived F(AB) fragments or F(AB)2-fragments norocea anti-virus antibodies were immunized rabbits to conventional circuits. Upon completion of the immunization of animals received hyperimmune serum.

From the received hyperimmune sera were isolated antiidiotype, specifically interacts with F(AB)-fragments norocea antibodies to antigens of viruses ABN using immunoaffinity sorption (conventional schemes) on columns with immobilized immunoglobulins mink, patients ABN.

To eliminate possible is with immobilized purified immunoglobulins from the serum of healthy mink.

Received antiidiotypic the drug is used as a prophylactic vaccine to prevent the disease ABN.

In some cases as a vaccine preparation was used F(ab) or F(ab)2-fragments of antiidiotype obtained by conventional circuits.

3. Preventive vaccination.

Prophylactic efficacy derived vaccines were tested on young dark brown mink. The experiments were carried out on the infected ABN populations of animals in the breeding animal-breeding sovkhoz "Codesense" Olonets district the Republic of Karelia.

For vaccination were selected animals without apparent clinical symptoms ABN. They all got 3 consecutive injections (at 10-day intervals between injections) one of the options vaccines. The drugs were administered subcutaneously, 2-3 mg of total protein, adsorbed on the suspension of the hydroxide of aluminum.

We used the following vaccine preparations (each option vaccination were tested on 50 animals; control group - 600 animals): - VS30 - solid antiidiotypic analogs of viral protein with a molecular mass of 30 KD; - VS80 - solid antiidiotypic analogs of viral protein with a molecular mass of 80 KD; - VS30+VS80 protein with a molecular mass of 80 KD; - VF80 - Fab-fragments antiidiotypic analogs of viral protein with a molecular mass of 80 KD.

Over the next 4 months compared levels of mortality among unvaccinated animals and burrows same population vaccinated with different drugs. Also evaluated the presence or absence of symptoms ABN animals visually and by necropsy (the conclusion of a veterinarian of animal-breeding sovkhoz). Examples (results) - see the table at the end of the text.

CONCLUSIONS: 1. Vaccine antiidiotypic drugs VF80 and VS80 effectively prevent the development of acquired immunodeficiency virus - Aleutian mink disease not less than 80-90% of vaccinated animals, and the most effective is the use of the drug VF80, presents Fab-fragments antiidiotypic analogs of viral protein with a molecular mass of 80 KD.

2. For example (experimental model) create a new way of prevention of Aleutian mink disease developed schematic diagram of the receiving and proven practical effectiveness of antiviral vaccines derived from antiidiotypic analogs of viral antigens. The developed method can be potentially effective and animals.

3. The use of preventive antiidiotypic vaccine Aleutian mink disease not less than 3-4 times to reduce morbidity and mortality of the animals and can be used effectively in the conditions of specialized fur farms affected Aleutian disease.

Claims

The way the specific prevention of Aleutian disease of mink, characterized in that conduct injecting the holes antiidiotypic antibodies or F (ab) fragments, which are analogues of antigen targets of causal virus.

 

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FIELD: clinical immunology.

SUBSTANCE: preparation represents 9.0-11.0% solution of horse blood serum immunoglobulins with pH 7.0-7.5 and content of γ-immunoglobulin fraction at least 90.0% of the total protein. Nonspecific fractions in preparation are albumin, α- and β-globulin fractions in amount not larger than 10.0%, and residual ethyl alcohol in amount not larger than 4.0%. Preparation is characterized by virus-neutralizing antibodies against virus Marburg. Titer of virus-neutralizing antibodies is at least 1:2048 and can be used for emergency prevention of Marburg fever in humans.

EFFECT: extended immunoglobulin application area.

1 tbl

FIELD: biotechnology, immunology, veterinary science.

SUBSTANCE: invention proposes the preparation that represents 9,0-11.0% immunoglobulins solution of horse blood serum with pH value from 7.0 to 7.5. The content of γ-globulin fraction in this preparation is 90.0%, not less, of the total protein amount. The preparation comprises albumin, α- and β-globulin fractions as nonspecific impurities in the amount 10.0%, not above, and residual ethyl alcohol in the amount 4.0%, not above. The preparation is apyrogenic, non-toxic and sterile.

EFFECT: valuable properties of preparation.

1 tbl, 1 ex

FIELD: immunology, biotechnology.

SUBSTANCE: invention describes antibody and its fragments neutralizing rabies virus and a method for treatment of patient subjected for effect of rabies virus by using indicated antibody and its fragment. Invention discloses variants of isolated nucleic acids encoding polypeptides carrying light and heavy chain of antibody, respectively. Also, invention describes expressing vector carrying at least one of indicated nucleic acids. Using this invention enhances span-life of patients after effect with rabies virus on them and can be used in corresponding prophylactic therapy of such patients.

EFFECT: valuable medicinal properties of antibody and nucleic acid.

14 cl, 1 dwg, 1 tbl, 2 ex

FIELD: veterinary science.

SUBSTANCE: the present innovation deals with immunization of down-calving cows with a live dry culture associated vaccine against paragrippe-3 (PG) and infectious rhinotracheitis in cattle at the dosage of 2 ml 1.5 mo before calving at simultaneous intramuscular injection of Selecor at the dosage of 10 mcg/kg animal's body weight; in 14 d one should introduce Selecor once more at the same dosage; then Selecor should be once injected intramuscularly for calves born in above-mentioned cows for 0.5-1 h after their birth. The innovation increases colostral cell and humoral immunity in calves, decrease sickness rate and accelerates the recovery.

EFFECT: higher efficiency of prophylaxis.

3 ex, 6 tbl

FIELD: veterinary science.

SUBSTANCE: one should immunize down-calving cows 1.5 mo before calving with a live, dry culture associated vaccine against paragrippe-3 (PG) and infectious rhinotracheitis in cattle (IRT) and simultaneously it is necessary to introduce Ligfol per 5 ml/animal; in 14 d Ligfol should be introduced once more at the same dosage; for calves born form these cows for 0.5-1 h after the birth it is necessary to introduce Ligfol once intramuscularly at the dosage of 2 ml. The innovation decreases the quantity of diseases and accelerates the recovery in animals.

EFFECT: higher efficiency of immunoprophylaxis.

3 ex, 6 tbl

FIELD: veterinary science.

SUBSTANCE: on proving the diagnosis one should introduce immune serum of animals-donors for sick calves till clinical recovery at the titers of hemagglutinins to IRT virus being 1:256, to PG-3 virus being 1:1280 and to VD-BC virus being 1:1024, and, additionally, it is necessary to apply a probiotic preparation lactobifadol at 32x108 microbial cells bifido- and 4x107 lactobacteria. The innovation increases the quantity of recovered animals, shortens the terms of therapy and the number of relapses.

EFFECT: higher efficiency of therapy.

2 ex, 3 tbl

FIELD: veterinary, immunology.

SUBSTANCE: healthy calves are immunized three time with 10-day interval by hypodermic injection of immune serum from donor animal with hemagglutinin titer to infective rhinotracheitis virus of 1:256; to parainfluenza-3 virus of 1:1280; and to viral diarrhoea virus of 1:1024. After the first injection additionally probiotic preparation lactobifadol in amount of 16x108 microbial bifido- cells and 2x107 lactobacteria for 10 days in three courses.

EFFECT: nocifensor activation; decreased calf morbidity rate.

3 ex, 5 tbl

FIELD: veterinary science.

SUBSTANCE: the suggested remedy contains alcoholic extract of the mixture of the equal parts of Echinacea and coltsfoot herb and inflorescences, flowers of small-leaved linden and licorice roots. Method for preventing and treating respiratory diseases in calves includes subcutaneous injection of hyperimmune serum that contains antibodies to the viruses of parainfluenza-3 and infectious rhinotracheitis in titers being 1:1280 and 1:256, not less, moreover, additionally for inner intake it is necessary to apply alcoholic extract of equal quantities of Echinacea and coltsfoot herb and inflorescences, flowers of small-leaved linden and licorice roots as a 7-8%-aqueous solution at prophylactic purpose at the dosage of about 1.5-2.5 ml/kg animal body weight for 15 d at interval of 12-24 h, at curative purpose - at the dosage of about 3.5-4.5 ml/kg animal body weight at 12-h-long interval till clinical recovery.

EFFECT: higher efficiency.

2 cl, 3 ex, 3 tbl

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