Cell line of calf kidney bos taurus rbt (rene bos taurus) for reproduction of animal viruses

FIELD: biotechnologies.

SUBSTANCE: permanent cell line of calf kidney Bos taurus RBT, deposited in 2010 into the Russian Collection of Cell Cultures (V) (Russian Academy of Agricultural Sciences, agricultural animals) under the No.74. The RBT line is produced from primarily trypsinized kidney cells of a 2-month calf in process of 65 serial passages in a single layer in the medium of 0.4 GLA on Earl with 10% foetal serum of cattle. Forms a population of epithelial cells, which is homogeneous by morphology, with considerable polymorphism in karyotype. At the same time population of cells with 51 chromosomes makes 32%, in contrast to normal karyotype Bos taurus (60 chromosomes), in RBT there are 9 metacentric and 1 submetacentric chromosomes, the number of polyploids is not more than 2%.

EFFECT: RBT line has high sensitivity to a corona virus, a virus of infectious rhinotracheitis, a plague virus and virus diarrhoea, and also high proliferative activity and ability for growth in rotary vessels.

4 dwg, 5 tbl, 4 ex

 

The invention relates to the field of biotechnology and Virology and the receipt of a new cell line kidney calf (Bos taurus), intended for reproduction animal viruses for the purpose of detecting, selecting, holding virological studies, as well as for the manufacture of diagnostic and preventive veterinary biologics.

Permanent cell line kidney of cattle (cattle) are widely used in scientific research and biotechnology [1, 2]. They are derived from embryos, newborn and adult animals [3]. Known cell line kidney and other organs of cattle:

- kidney calf - MDBK - Madin-Darby Bovine Kidney cells;

- kidney calf - Taurus - 1;

- kidney cattle - PT-80;

- kidney embryo bull - AUBEK;

- trachea embryo bull - FBT.

These and other cell lines of calf kidney and other organs of cattle are used for cultivation of virus diarrhea, bovine rhinotracheitis [4], influenza, parainfluenza III, coronaviruses, adenoviruses and other (A.S. USSR №1347448, 1985; A.S. USSR №1212043, 15.10.1985 [5]). Cell lines differ in index of cell proliferation, adhesion, sensitivity to viruses, their ability to reproduce in one way or another nutrient medium, antigenicity, cytogenetic characteristics, morphology. Permanent cell line derived from the kidney and other organs of cattle, divided according to the article the penalties chromosomal transformation into 2 groups: the first group with allodiploid set of chromosomes (58-62 chromosomes) and without significant chromosomal rearrangements, which includes cell line Taurus-1, PT-80, AUBEK [5, 6, 7]; the second group with hypoploidy modal class (50-52 chromosomes) and the presence of from 8 to 11 marker, transformed chromosomes, which includes cell line MDBK, FBT [8, 9].

During the long passage above permanent cell lines, usually heterogeneous cytogenetic, morphological, and physiological characteristics that are of limited use in virological work that requires extensive Arsenal of animal cells by obtaining new cell lines with stable cultural properties and a wide range of applications in the production of vaccines.

The closest cell line by cytomorphological, karyological and cultural characteristics is MDBK, which was obtained in 1958 [8] and has several tratariamos [9]. On biotechnological parameters, it is inferior to newly acquired line: average productivity MDBK with Klin mattress 100÷130 million cells, with the roller volume of 3 l 120÷150 million cells. Also, this cell line is rapidly eliminated after reducing serum less than 5%.

The problem to which this invention is directed, was the receipt of a new line of cells from the kidney of veal with improved biotechnological characterized the ticks, suitable for detection and selection of a wide range of viruses, but also for the production of diagnostics and specific prophylaxis of viral diseases of animals.

The problem is solved by obtaining a cell line kidney calf (Bos Taurus) RBT - Rene Bos Taurus.

The essence of the invention is illustrated in the following graphics:

Figure 1 - Metaphase plate cell lines RBT - 51 chromosome;

Figure 2 - Ideograma karyotype RBT;

Figure 3 - Monolayer cell line RBT in the logarithmic phase of growth;

4 is a Diagram of the spectrum of LDH isoenzymes primary culture (FRI) and permanent crops (RBT).

Obtaining cell line kidney calf (Bos Taurus)

RBT - Rene Bos Taurus

We received a new line of cells RBT from primary trypsinization kidney 2-month-old calf. The monolayer cells primary cultures of kidney calf was formed on the fifth day after sowing primary trypsinization cells at a concentration of 300,000 cells in 1 ml (with a change of medium after 2 days of cultivation). For growing primary cultures were used nutrient medium on the basis of the saline Earl with 0.4% hydrolyzed lactalbumin (Peptone from lactalbumin ensimatic, readilly soluble "Serva"), 0.3 g/l glutamine and vitamins, with the addition of 10% fetal cattle serum. After the first subculture remained significant potency to proliferation of the t is to the first passage monolayer was formed on day 3 or 4 when seeding concentration of 150,000 cells per milliliter. Initial passages of subculture were only on the environment of 0.4% GLA at the Earl with the addition of 10% fetal serum. Synthetic and semi-synthetic environments (Wednesday Needle MEM and SRP) has been a sharp decrease in the mitotic activity of the cells. Until the 30th passage of the cultivation was carried out on the environment of 0.4% GLA at the Earl. Until the 11th passage subculture kidney calf differed polymorphism: attended fibroblast-like, epithelioid, spindle-shaped and polygonal cells. Productivity was insignificant - 15÷20 million cells Klin mattress. Sieving was performed once a week in the proportions 1:2, 1:1.

To 30-th passage appeared a tendency to isomorphism and increase growth activity subculture: fibroblast-like cells began to grow in size and eliminirovat. There were groups epitheliopathy cells of small size (up to 15 µm in adhered condition), which were actively growing in large colonies and replaced the fibroblast-like cells, large cells. 10÷15% of the population of cells began to appear 1-2 metacentric chromosomes (except the X chromosome). To the same period began to increase productivity subculture (up to 20÷25 million cells Klin mattress). After each subculture epitheliopathy cells and fibroblast-like cells were enlarged and destroyed under the action of the trypsin. With this clause the period cultured cells may have adapted to environments Needle MEM and SRP. One condition remained unchanged - the use of fetal cattle serum, free from Mycoplasma and latent viruses.

The following detailed testing subculture held at 50-52 passages. To this period the culture was fully adapted to the environments Needle, SRP, MEM; productivity of cells with Klin mattress increased to 50 million Cell population predominantly consisted of epitheliopathy (polygon) cells, fibroblast-like cells almost completely eliminated, but there was a giant cells (up to 10% of the total area of the monolayer), which remained in the passages. There have been significant changes in the karyotype: appeared 6-7 meta - and submetacentric autosomes; the modal class was formed within 53-54 chromosomes, poliploids and giant cells was 6÷8%.

Further study of the culture was carried out at 65-m passage, when the productivity of cells with Klin mattress grew to 100 million or more, and sieving cells could be carried out in the proportions 1:4 to 1:8. Also was revealed the ability of cell lines to growth in a rotating 3-liter vessels.

Formed the population of cells at this stage of its existence, on the above grounds, appeared as a regular (human) cell line. Karyological study of cells 65 and 81 passages showed that mo is social class has stabilized at a level 51 chromosomes, there were 10 of meta - and submetacentric chromosomes, which are markers of a new cell culture (figure 1, 2).

The cells formed a monolayer are isomorphism epitheliopathy and fusiform cells, the number of poliploids (giant cells) decreased significantly and amounted to less than 1%. Cell nuclei mainly rounded, contain 1-2 large nucleolus (Figure 3). In the stationary growth phase cell membrane weakly kontroliruyutsya (formed pseudointimal). At the same time, the cell culture well trypsinized to individual cells.

The analysis of the LDH isoenzymes confirmed the species origin of this cell cultures from cattle Bos taurus (Figure 4). Range lines of LDH isoenzymes diagram for primary cell culture PT (kidney calf), are identical for a new transplantable cell culture from kidney calf - RBT.

The sowing of backside at all stages of the formation of cell cultures showed no contamination with bacteria, yeast, fungi and Mycoplasma.

Cells underwent a total of 65 passages to the transformation from a subculture in a highly productive continuous cell line with the above properties. Their samples on 67-94 passages, were laid stored in the Cryobank of the fgbi "ARRIAH" under the copyright on the requirement Line cells of the kidney calf (Bos Taurus) RBT Rene Bos Taurus".

Sample permanent cell line kidney calf (Bos Taurus) RBT - Rene Bos Taurus selected passage 78 deposited in the 2010 collection of somatic cells of agricultural and game animals the all-Russian research Institute of experimental veterinary RCCC(P) (she RAAS) under No. 74.

Biotechnological characteristics

Cells RBT grown at pH 6.9÷7.2 and a temperature of 36÷38°C in a monolayer, in plastic or glass containers with a capacity from 50 to 1500 ml and in rotating vessels 2000-3000 ml at a ratio of environment and volume of the vessel in the range 1:6 to 1:10. While using MEM medium with 0.25% hydrolyzed lactalbumin or SRP salt solution Earl with the addition of 10% serum of cattle. Table 1 presents data on the productivity of cell cultures RBT under cultivation in different culture vessels.

With the multiplicity of sieving 1:4÷1:8 monolayer formed after 2-4 days of cultivation without changing the environment. Subcultivation is a common method, causing the dispersion of monolayer mixture, containing 0,02% of versene (ethylenediaminetetraacetate) and 0.25% trypsin in a 1:1 ratio. The percentage of viable cells determined by the method of electron microscope staining removed suspension - Trifunovi blue, and monitoring of the growth dynamics of seed culture. The kinetic nature of the stick line RBT 94 passage showed the maximum harvest level 150÷200 million cells in the mattress 1500 ml is provided in 4 days with seeding density of 100 thousand cells per ml (table 1). Productivity in roller containers reaches 400 million/CL In terms of size, productivity 1 cm2over 660 thousand cells.

In standard culture conditions cell line has a high level of glycolysis. When the acidification of the environment, degeneration of cell culture begins at 5÷7 days. The complete destruction of the monolayer at 37°C comes on 10÷12 days of growth.

In addition to the standard parameters of cultivation, cell line RBT steadily growing for more than 10 passages with the use of 2% cattle serum instead of 10%. The decrease in cell proliferation occurs after 5 consecutive passages with the use of 0.6% whey.

Ampoules with cells RBT, concentrated to a density of 5.0÷10,0×106cells/ml in fresh growth medium with 10% dimethyl sulfoxide, and stored in liquid nitrogen. Cells are thawed, Ottawa vials in a water bath at 37 ° ÷38°C. the Optimum level of proliferative activity of a cell culture marked before freezing, was achieved at 2 passage after thawing.

The monolayer formed by the cells RBT, is characterized by an ordered orientation of elongated polygonal epitheliopathy cells. Cell nuclei, as PR is usually round or oval shapes, various sizes (depending on the stage of the cell cycle), with 1-2 large and several small nucleoli.

In the process of getting constantly cell lines RBT species was tested methods and karyological analysis 11, 30, 50, 65, 81 passages. In all cases, the spectrum of LDH isoenzymes were detected signs of the species Bos Taurus. In the karyotype of the cell line occurred stabilization: formed the modal class of the population of cells with 51 chromosomes (32%); cells 50-52 chromosomes constitute 77%; polymorphism from 40 to 65 chromosomes. In fetuses present 9 metacentric and one submetacentric chromosome, Y-chromosome is eliminated, polypoids less than 1%.

When examining cells cytochemical methods (staining with acridine orange or olivomycin), as well as when sowing on backside (BCH, MPA, TGS and 0.3% PPLO agar), in the study by electron microscopy and PCR, the presence of bacteria, fungi and mycoplasmas not found.

Cells RBT are designed to highlight and accumulation of animal viruses to determine their infectious activity, the study of genetic and phenotypic properties, as well as for the manufacture of diagnostic and vaccines, including viruses or their components. These cells susceptible to infection with viruses of different families infe the operating rhinotracheitis (SEM. the herpes viruses), plague (SEM. paramyxoviruses) and coronavirus (SEM. coronaviruses) cattle, viral diarrhea is a disease of the mucous cattle (SEM. flavivirus).

The essence of the invention is illustrated by examples of its use, which do not limit the scope of the invention.

Example 1.

Reproduction of coronavirus cattle in cell cultures

Cell suspension concentration 50÷150 thousand cells/ml were scattered in stationary or rotating vessels. Cultivation of the cells was performed in the environment of the Needle from 0.25% GLA or SRP with 10% serum of cattle at a temperature of 36 to 37°C at pH of 6.9÷7,1 without changing the environment to form a full dense monolayer. For virus used a cell monolayer, no signs of contamination and degeneration.

The infected culture was performed by contact of concentrated suspensions of virus in monolayer Klin mattresses, after removing the used growth medium and double rinsing the cells with warm Hanks solution. The contact of the virus with the cells was carried out for 1 hour with 5÷8 ml of viral suspension at 37°C, added in an amount to provide a multiplicity of infection of 1.0 to 5.0 TCD50/ml per cell. This portion of the virus evenly watered monolayer, gently shaking the culture vessel. To ensure the indicated multiplicity of infection concentrate seed VIR is sa and supportive environment mixed, as a rule, in the ratio of 1:30 to 1:50. Then in the culture vessels were added Wednesday SRP without serum and incubated at pH 7,2÷7,3 within 45÷96 hours at 37°C until complete lysis of the infected culture.

Thus obtained viral material was frozen at a temperature of minus 40°C to extract the maximum virus yield. Then it was titrated in cell culture RBT and Taurus-2 in penicillin vials. The titer of the virus obtained from the culture of RBT was within 8,0÷9,0 lg TCD50/ml (table 2).

In subsequent experiments it was found that infection of cells RBT at any level after 65 passage of the coronavirus cattle his crops consistently played with titers of infectious activity, were in the same range.

Example 2.

The reproduction of the virus of infectious bovine rhinotracheitis cattle

Cells and virus were treated as described in example 1. The titration was carried out on cell cultures RBT, MDBK, ADC-04. Virus reproduction in cell culture RBT was 24÷48 hours, faster than in other cultures. The titer reached 7,0÷8,0 lg TCD50/ml (table 3).

Example 3.

Reproduction viral diarrhea is a disease of the mucous cattle

Cells and virus were treated as described in example 1. The difference was that when the contact of the virus with the monolayer of cells in the environment in RosNOU suspension was added trypsin at a concentration of 0.01%. The titration was carried out on cell cultures RBT and Taurus-2. The titer of infectious activity in the cell culture RBT reached 6,0÷7,0 lg TCD50/ml and 1:100 in ELISA (table 4). The proliferative activity of the cell culture RBT in 2-3 times above, than at Taurus-2, therefore, the production of vaccines based on it more profitable.

Example 4.

Reproduction of rinderpest virus of cattle

Reproduction of rinderpest virus of cattle occurs within 96÷120 hours. For the cultivation of the virus used the environment of the SRP with 2.5% serum of cattle. The titration was carried out on permanent cell lines RBT, Taurus-2 and subculture FRI. The titer of infectious activity during the cultivation of the virus in cell culture RBT reached 7,0 lg TCD50/ml (table 5).

Studies on the cultivation of coronavirus and a virus of cattle in cell lines RBT indicate a high sensitivity of this culture to the above viruses - 8,0÷9,0 lg TCD50/ml. In the case of a virus of cattle cells RBT was a convenient system for titration of the virus.

A comparative study of the reproduction of coronavirus, virus, rhinotracheitis, diarrhea, and cattle plague on different crops allow us to conclude that the proposed new continuous cell line RBT (derived from kidney calf) has a high sensitivity to the above viruses and, in combination with high proliferat the main activity and the ability to grow in rotating vessels, makes it suitable for the production of prophylactic antivirals.

Sources of information

1. Syurin V.N., Samuyilenko YA, Soloviev BV, Fomin, NV - a Viral disease of animals. M: Spa (scientific production Association and BP, 1998.

2. Spier R.E., Griffiths JB - Biotechnology animal cells. M: Agropromizdat, 1989. - 185 S.

3. Deacons L.P., Shitikov VI - Animal cell in culture. Methods and their application in biotechnology. M: Sputnik+, 2009, 656 C.

4. Kozyrenko TI, deacons, L.P., Pozdnyakov A.A. study of the sensitivity of transplantable cells to the virus of infectious bovine rhinotracheitis cattle // Scientific basis of the technology of industrial production of veterinary drugs: proc. Dokl. the second vsesojuz. proc. M. 1981. - P.20-21.

5. The CATALOGUE. The specialized collection transplantable somatic cell cultures of agricultural and game animals RCCC(P), (she RAAS), Moscow, 2006, 115 S.

6. Mironov L.L., Transfiguration NICHOLAS, Shitikova G.S. and other Domestic line transplantable cells of the kidney calf - substrate for biotechnology - Biotechnology, 1998, No. 5, s-31.

7. Mironov L.L., Popova E, Conosco I., Chapaev E, Zybin D.V., Hakobyan, AS the Experience of the Bank copyright lines transplantable cell lines and their application in Virology practice - Biotechnology, 2000, No. 6, P.41-47.

8. S.H. Madin, N.B. Darby Established Kidney Cell lines of Adult Bovine and Ovine Origin // Proc. Soc. Exp. Biol. Me. - 1958. - Vol.98. - P.574-576.

9. The CATALOGUE. Russian collection of cell cultures, Ed. by Gpeni, St. Petersburg, 2004, 315 S.

Table 1
Dynamics of cell proliferation cell lines RBT in different culture vessels
Conditions for growing cells in a monolayerThe seeding concentration (million/ml)Time of cultivation (days)The productivity of the cells from the culture vessel (million)The rate of increase
Glass culture vessels (Klin mattresses) with a volume of 1500 ml0,1297,0±0,4the 3.8
3120,0±0,14,8
4200,0±0,18,0
0,05255,0±0,24,2
390,0±0,17,0
120,0±0,29,6
0,0252--
3--
460,0±0,19,8
Rotating vessels with a volume of 3000 ml0,12150,0±0,25,0
3200,0±0,56,6
4350,0±0,4the 11.6
0,152180,0±0,24,0
3250,0±0,15,5
4400,0±0,38,8

Table 2
Reproduction of coronavirus cattle in cell cultures
Cell cultureTime of cultivation (h)The titer of infectious activity (lg TCD50/ml)Electron microscopy (particles per ml)
MDBK96-1202,0-3,0106,5-7
Taurus-296-1203,0-4,0107-7,5
RBT (Klin mattresses)40-486,0-7,0106,5-7
RBT (roller)40-488,0-9,0108-8,5

Table 3
The reproduction of the virus of infectious bovine rhinotracheitis cattle in cell cultures
Cell cultureTime of cultivation (h)The titer of infectious activity (lg TCD50/ml)
ADC-0448-727,0-8,0
MDBK72-807,0
RBT24-487,0-8,0

Table 4
Reproduction viral diarrhea - diseases of the mucous membranes of cattle in cell cultures
Cell cultureTime of cultivation (h)The titer of infectious activity in ELISAThe titer of infectious activity (lg TCD50/ml)
Taurus-2Lasts for 48-961:100-1:5007,0
RBT (Klin mattresses)96-1201:1006,0
RBT (roller)Lasts for 48-961:1006,0-7,0

Table 5
Reproduction of rinderpest virus of cattle in cell cultures
Cell culture Time of cultivation (h)The titer of infectious activity (lg TCD50/ml)
PT (primary culture of renal calf)72-96of 5.75 to 6.0
Taurus-472-966,0-6,5
SPAW96-1207,0-7,5
RBT96-1206,0-7,0

Continuous cell line kidney calf Bos taurus RBT deposited 2010 RCCC(P) (she RAAS) under No. 74, for reproduction animal viruses, characterized in that it is:
- obtained from primary trypsinization kidney 2-month-old calf in the process 65 serial passages in monolayer on the environment 0,4 CHAP on Earl with 10% fetal cattle serum;
- forms a homogeneous morphology of a population of epithelial cells with significant polymorphism in the karyotype, while the population of cells with 51 chromosomes is 32%, in contrast to the normal karyotype Bos taurus (60 chromosomes), RBT present 9 metacentric and 1 submetacentric chromosomes, the number of poliploids not more than 2%;
- under cultivation in the monolayer cyclic method provides a reproduction of karanvir the sa cattle with a title 8,0-8,5 lg TCD 50/ml of a virus of cattle with a title 7,0-8,0 lg TCD50/ml, rinderpest virus of cattle with a title 6,0-7,0 lg TCD50/ml titer infectious activity of viral diarrhoea of calves in ELISA reaches 1:100.



 

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SUBSTANCE: invention relates to biotechnology, genetic engineering and virology. Described is a chimeric protein comprising an L2-epitope of a human papillomavirus (HPV) type-16 inserted in a loop region of a human papillomavirus type-16 L1 protein; and a capsid which is an aggregate of the chimeric protein. The loop region to which the L2-epitope is to be inserted is located between an amino acid residue at position 430 and an amino acid residue at position 433. The L2-epitope has an amino acid sequence represented by any one of the following formulae: LYKTCKQAGTCPPDIIPKVEG (18-38 L2-epitope); GGLGIGTGSGTGGRTGYIPL (56-75 L2-epitope); and DPVGPLDPSIVSLVEESSFI (96-115 L2-epitope).

EFFECT: invention can be used to obtain a vaccine against human papillomavirus.

18 cl, 7 dwg, 5 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: chimeric flavivirus contains at least one mutation in a coat protein of chimeric flavivirus, and one or more mutations in (i) 3'-untranslated region (3'-UTR) of a chimeric flavivirus genome and/or (ii) a capside protein of chimeric flavivirus.

EFFECT: chimeric flavivirus shows reduced viscerotropism as compared to mutation-free chimeric flavovirus; virus is completely attenuated.

32 cl, 8 dwg, 7 tbl

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science. A method for immunoprophylaxis of respiratory diseases in calves involves immunisation with an associated vaccine for parainfluenza-3, infectious rhinotracheitis and bovine viral diarrhoea viruses in a combination with the use of an alcoholic solution of a herbal raw material as an agent for non-specific prophylaxis with the herbal raw material presented by a mixture of equal proportions of purple Echinacea herb and blossom clusters, Syrian rue herb, tillet blossom and common licorice roots which is imbedded in 70% ethanol in relation 1:10, kept in a dark place at temperature 18-20°C for 7 days; the prepared tincture is prescribed orally in the form of 7-8% aqueous solution 1.5-2.0 ml/kg of animal's body weight for 15 days with immunising calves of 60 days old.

EFFECT: invention provides higher effectiveness of immunoprophylaxis.

4 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: method involves isolation, culture and replication of avian embryo stem cells in a complete culture medium containing all growth factors in the presence of an inactivated feeder layer, and added with animal serum; separation of the growth factors, serum and feeder layer; production of the non-adhesive embryo stem cells; proliferation of the produced cells in a suspension in the form of cell balls; infection of said cells with said virus; addition of the second serum-free medium to the cell culture; further culture of the infected cells for virus replication and collection of said virus.

EFFECT: invention is applicable for preparing vaccines and particularly enables producing viral strains which cannot be adequately growth in eggs.

38 cl, 23 dwg, 4 ex

FIELD: medicine.

SUBSTANCE: strain is recovered from a patient taking a specific chemotherapy for 3 years, has one basic and four secondary mutations which can change chemopreparation sensibility. The strain is deposited in the State Collection of Viruses on the basis of Federal State Government-Financed 'Ivanovsky Research Institution of Virology' of Ministry of Health and Social Development of the Russian Federation, No. GKV No.1033.

EFFECT: presented strain may be used as a biotechnogical natural model for the purpose of developing the diagnostic and vaccine preparations, and for screening of various compounds and drug test.

5 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology, virology and veterinary. An attenuated recombinant classical swine fever virus (CSFV) is described. Modification is carried out by progressively mutating a portion of the E2 gene of the highly pathogenic strain Brescia. As a result, two to six amino acids on the section 829-837 are replaced with two to six amino acids which are characteristic for the heterologous E2 glycoprotein of the Bovine Viral Diarrhea Virus (BVDV). A classical swine fever vaccine is also obtained based on the obtained attenuated CSFV. The invention can be used in veterinary.

EFFECT: classical swine fever vaccine is obtained.

7 cl, 7 dwg, 4 tbl, 7 ex

FIELD: biotechnology, veterinary science.

SUBSTANCE: invention relates to therapeutic vector used in therapy of infectious diseases in cats that comprises at least one foreign nucleic acid each of that (a) encodes protein taken among the group consisting of feline protein CD28 represented in SEQ ID NO:8 or its immunogenic moiety; feline protein CD80 represented in SEQ ID NO:2 or 3, or its immunogenic moiety; feline protein CD86 represented in SEQ ID NO:6 or its immunogenic moiety, or feline protein CTLA-4 represented in SEQ ID NO:10 or its immunogenic moiety; and (b) nucleic acid that is able to be expressed in insertion of vector in the corresponding host. Indicated therapeutic vector is used in effective dose as component of vaccine against infectious diseases in cats for their immunization and in methods for enhancement or inhibition of immune response in cats and reducing or eradication of tumor in cats. Invention provides stimulating the activation and proliferation of T cells and to enhance effectiveness of control of infectious diseases in cats.

EFFECT: valuable biological properties of recombinant virus.

41 cl, 13 dwg

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