RU1692022C - Foot-and-mouth disease vaccine adjuvant - Google Patents

Abstract

FIELD: veterinary medicine; immunology. SUBSTANCE: polyglycerol alkanoates mixture produced by using unsaturated fatty acids as the base is applied as VNIIZh emulsifier. The following proportions were taken in by mass: VNIIZh emulsifier -10-15, oil base 85-90. EFFECT: improved immunostimulating effect of the adjuvant, lowered viscosity and solidification temperature of the preparation produced by using the adjuvant as the basic component. 4 tbl

Description

 The invention relates to veterinary virology, and more specifically to oil adjuvants, and can be used in research and biotechnology industry in the development and production of oil adjuvants, and based on them immunizing preparations.

 The aim of the invention is to increase the immunostimulating activity of the adjuvant, lowering the viscosity and pour point of the immunizing preparation obtained on its basis.

PRI me R 1. For experimental verification of the inventive adjuvant prepare seven mixtures of ingredients, three of which showed optimal results (table 1)
The optimal composition of the proposed adjuvant is determined by studying the physical properties of the immunizing emulsions from foot and mouth disease virus A ₂₂ obtained on its basis. Emulsions are obtained by mixing the aqueous (antigen) and oil (adjuvant) phases in a 1: 1 ratio in a tissue grinder PT-2 for 5 minutes at 5000 rpm.

As the oil base in the manufacture of the proposed adjuvant use mineral oil TU 3810188-84. As the aqueous phase, FMD virus antigen A _{22 is used} , prepared in accordance with the Provisional Instructions for the Production of Bivalent Type A and O Vaccines against Foot and Mouth Disease Control (from lapinized virus).

 Information about the stability, viscosity and fluidity of immunizing emulsions using various adjuvants are also given in table 1.

Studies have established that the following compounds of the proposed adjuvant containing, by weight, possess optimal properties. Emulsifier ARRIAH 10-15 Oil base 85-90
Examining samples of emulsion immunizing preparations prepared using various formulations of the proposed adjuvant. It was found that when the ratio of adjuvant ingredients (wt.) 80-82.5: 17.5-20 immunizing preparations have a high viscosity (more than 500 cm ³ / s). Emulsion preparations based on adjuvants containing less than 10% of the VNIIZh emulsifier (samples 6 and 7) are not stable, since they do not pass the generally accepted stability test for emulsion veterinary preparations.

 Similar results are obtained when using other oil bases: medical vaseline oil, perfume oil, vaccine oil, etc.

 The adjuvant is prepared as follows.

Take a sample of the emulsifier VNIIZH and dissolve in mineral oil in optimal proportions. The resulting solution was sterilized by known methods, either by heating in an oven to 120-130 ^{° C} for 2 hours, or by sterilizing filtration using a plastic marks "SF", EKS, UAM-150, etc. at 40-50 ^about C.

 The finished adjuvant is a clear, oily yellow to brown color liquid depending on the concentration and quality of the emulsifier. The adjuvant is poured into sterile vials and stored at room temperature until use.

The effectiveness of the proposed adjuvant in comparison with the prototype was tested in several experiments in the composition of emulsion FMD vaccines A ₂₂ .

PRI me R 2. Experiment emulsion FMD vaccines from the virus of type A ₂₂ based on various oil adjuvants. The values of fifty-percent immunizing doses (BMI ₅₀ ), determined 21 days after vaccination of white mice with control infection, are presented in table 2.

 From table 2 it is seen that the preparations containing the proposed adjuvant have a higher immunogenic activity compared with preparations containing the adjuvant (prototype) and incomplete Freund's adjuvant.

In the next series of experiments, the immunogenic activity of FMD vaccines from the lapinized virus A ₂₂ based on various oil adjuvants is studied in dynamics: respectively, 7.14 and 21 days after vaccination. The results of the experiment are presented in table.3.

From table 3 it follows that vaccines containing the proposed adjuvant have a more pronounced immunogenic activity than vaccines with an analogue and prototype both on the 7th day after vaccination, and on the 14th and 21st days. Only on the 7th day, the IMD value of _{50 of the} drug based on the proposed adjuvant is higher than that of the vaccine based on the incomplete Freund's adjuvant (IMD _{50 of} 0.23 ml versus 0.13 ml, respectively).

Example 3. For a comparative assessment of the immunogenic activity of FMD drugs from lapinized virus A ₂₂ , three series of emulsion vaccines are prepared: on the basis of the proposed adjuvant (series 405), the prototype adjuvant (series 403) and incomplete Freund's adjuvant (404). Control infection is carried out 10 days after vaccination. Vaccines are administered at a dose of 1 ml intramuscularly. 8 days after the challenge, the animals are sacrificed and the reactogenicity of the vaccine is evaluated by opening the injection site. The results of the experiment are presented in table.4.

 During the experiment, all animals looked clinically healthy.

 Thus, the data in table 4 indicate that the proposed adjuvant has pronounced immunostimulating properties, and the apparatus based on it has a low reactogenicity.

Claims (1)

 ADDJUVANT FOR VACCINE AGAINST FMD, containing an oil base and an emulsifier, characterized in that, in order to increase the immunostimulating activity, lower the viscosity and pour point of the immunizing preparation obtained on its basis, the VNIIZ emulsifier is used as an emulsifier, which is a mixture of non-polyglycerol-based alkanoic fatty acids acids in the following ratio of components, wt. Emulsifier ARRIAH 10 15
Oil base 85 90

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