RU2067002C1 - Living cultural vaccine against carnivorous plague - Google Patents

Abstract

FIELD: veterinary. SUBSTANCE: vaccine strain "ЭПМ" of carnivorous plague virus is used for preparing living cultural vaccine against carnivorous plague. Strain is grown on Eagle medium with KPC serum or with foetal serum and antibiotics. Repeated virus growing is carried out on serumless medium that ensures to decrease concentration of heterologic proteins up to 0.3 mg/ml "ВСЖ". 0.7 vol. p.p. "ВСЖ" at biological virus activity 10^3,7-0,2ТЦД_50/мл is mixed with 0.3 vol. p.p. stabilizing agent that is a mixture of sorbitol, polyglucine and gelatose (0.05; 0.05 and 0.2 vol. p.p., respectively). Vaccine is poured into ampoules or small bottles (by 0.5 ml) and lyophilized. Vaccine can be used for prophylaxis against carnivorous plague. EFFECT: improved method of vaccine preparing. 2 tbl

Description

 The invention relates to veterinary medicine and can be used to prevent carnivore plague.

Known live culture vaccine against carnivore plague containing the virus-containing liquid of the vaccine strain "EPM" of the plague virus with biological activity of 10 ^3.5 TPD _{50 / ml} and a stabilizer sorbitol and gelatin [1]
However, the known vaccine contains a high concentration of heterologous proteins (about 7 mg / ml), which causes adverse reactions in animals after vaccination. In addition, the peak of immunity tension is reached on the 30th day.

The objective of the invention is to develop a vaccine with a reduced concentration of heterologous proteins (up to 0.3 mg / ml of virus-containing liquid) and biological activity of the virus 10 ^3.7 ± 0.2 TTCD _{50 / ml} , providing long-term intense immunity. In addition, as a stabilizer, a mixture of sorbitol, polyglucin and gelatose taken in vol. hours 0.05, 0.05 and 0.2 to 0.7 vol. including virus-containing fluid (VSL).

 The technical result obtained by solving this problem is expressed in reducing the reactogenicity of the vaccine and increasing immunity after vaccination.

 Example 1. Obtaining a vaccine.

To prepare the vaccine, a single-layer trypsinized tissue culture of Japanese quail embryo tissue cells obtained in roller units at a rotation speed of 2 rpm is used. Trypsinized cells are suspended in a growth medium consisting of Eagle medium supplemented with 8% cattle serum or 2% fetal serum and B-lactamase antibiotics. The suspension is poured into roll bottles and the virus is introduced in a dose of at least 100 thousand TCD _{50 / ml} per bottle.

Incubation of the virus-infected cell culture is carried out in roller units at 35 ^{° C.} at a rotation speed of 2 rpm. 5 days after the formation of the monolayer, the medium is drained and fresh growth medium without serum is added. At 8 days after infection, VSL is collected in vials. To all sterile individual viral collections with biological activity of the virus 10 ^3,7 TCD _{50 / ml} add a stabilizer at the rate of 0.7 vol. hours VSJ and 0.3 vol.h. stabilizer. A mixture of sorbitol, polyglucin and gelatose is used as a stabilizer based on their content of 0.3 vol.h. 0.05, 0.05 and 0.2 vol.h. respectively.

 After the vaccine is poured into ampoules or vials, it is subjected to lyophilization and control for sterility, antigenic activity and specificity for CPD and PFU, harmlessness and immunogenicity. One ampoule contains one vaccination dose of 0.5 ml vaccine.

 Example 2. Testing a vaccine.

 The vaccine was tested on 300 thousand thorsophrenics. This type of animal was chosen as the model as the most sensitive to this infection compared to other animal species (foxes, arctic foxes, etc.), which allows more reliable data to be obtained.

 As a result of the examination of immunized thorzofrets, data were obtained indicating the absence of pathological reactions due to a decrease in the concentration of heterologous proteins in the vaccine to 0.3 mg / ml VSL, i.e. vaccine areactogenic.

 In the serum of vaccinated animals, antibodies appear on the 7th day after vaccination. The neutralizing activity of antibodies is studied in the neutralization reaction on a primary culture of Japanese quail embryo cells. The results are presented in table 1.

 In all immunized animals, neutralizing antibodies to the carnivorous plague virus were detected, i.e. almost 100% seroconversion is observed.

 However, antibody titers indicating the presence of seroconversion in animals do not always guarantee the completeness of immunity, i.e. the immunity of animals to subsequent infection with a field virulent virus. Therefore, the evaluation of the vaccine only in terms of antibody titer is incomplete and, as a rule, should be supplemented by direct protection.

 Immunogenic activity was studied on thorsophrenics when they were infected subcutaneously with the virulent Snyder Hill virus. Vaccinated animals showed no signs of disease after the introduction of the virulent strain of the virus, while control animals died in the presence of a typical plague clinic. The research data are given in table.2.

 Thus, the resulting vaccine has a specific immunogenic activity, reduced reactogenicity and provides long-term intense immunity. TTT1

Claims (1)

Live culture vaccine against carnivorous plague, containing the virus-containing liquid of the vaccine strain "EPM" of the virus of the carnivorous plague virus with an admixture of heterologous proteins, sorbitol and gelatin, characterized in that it additionally contains polyglucin, the virus-containing liquid with a concentration of heterologous proteins up to 0.3 mg / ml of virus-containing liquid and biological activity of the virus 10 ^3.7 ± ^0.2 TCD _{50 / ml} in the following quantitative ratio of components, vol.h. Virus-containing liquid of the vaccine strain "EPM" of the plague virus with a concentration of heterologous proteins up to 0.3 mg / ml of the virus-containing liquid with biological activity 10 ^3.7 ± ^0.2 TCD _{50 / ml} 0.7
Sorbitol 0.05
Polyglukin 0.05
Gelatose 0.2

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