SU704183A1 - Method of preparing killed blue purulent polyvalent corpuscular vaccine - Google Patents

Description

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The invention relates to the field of medical industry, namely, to the production of vaccine npenapas.

A known method for producing a killed Pseudo-Pseudo-Pseudompna lerug Lnosa vaccine strains by separate Pseudompna lerug Lnosa vaccine strains, followed by their mixing tlj

However, a vaccine obtained in a known manner does not have a high protective property and the duration of the immunity being created does not exceed 2-3 weeks.

The purpose of the invention is to improve the protective properties and duration of the created immunity.

This is achieved by growing shafy Pseudomonas aferugtnosa R 1311, No. 1312, 1313, 1314 ,. 1315, 1316 and No. 1317.

The method is carried out in the following manner.

Ps.aeruginosa vaccine strains are grown separately on various dense nutrient media (m sappeptone agar, Hottinger agar, synthetic media with casein peptone, etc.) for 13-24 h.

From these cultures, suspensions containing 1 billion microbial cells in 1 ml (according to the optical standard of turbidity TSGNKI or spectrophotometer SF-40) in a physiological solution or bactericidal Gorgiyev liquid (ekteritsid) are prepared. Then, for the preparation of a polyvalent vaccine, suspensions of all 7 strains are made in equal volumes. In the case of preparing a vaccine with physiological saline, a preservative is added to the mixture (0.05% mertiolate or 0.5% formalin). Thus obtained vaccine

5 hold in a thermostat, at 37 ° C for 18-24-h, after which it is produced. Control of sterility, apyrogenesis, pathogenicity for experimental animals (white mice) and immunogenicity.

The corpuscular body killed in this way, a polyvalent blue-pus pneumonic vaccine, must have the following medico-biological properties: sterility) apyrogenicity, toxicity for those who, when administered 0.5 ml under the skin, immunogenicity, which is expressed in the 80100% survival rate of animals (infected as part of

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vaccines / 5-7 days after vaccination.

To test the immunogenic properties, the vaccine was administered to white mice weighing 18-20 g subcutaneously once and twice with a weekly interval in a dose of 0.5 MP, followed by testing of its protective activity. Infection of animals Carried out 7 days after the last vaccination: for this purpose, each of the strains of Pseudomonas aeruginosa that are part of the vaccine, as well as strains of serotypes | 011 and 09 are administered to animals intravenously at a dose of 0.5-10 microbial cells / ml, causing 100% death, {control animals (without prior vaccination). To check the duration of the protective effect of the vaccine, immunized animals are infected with a culture of Pepurus coli after various periods Last vaccination: on the 7th, 21st and 56th day.

The effect of the protective effect of the vaccine is evaluated on the basis of calculating the percentage of surviving immunized animals after infection with homologous strains of the Pyloric Syndrome.

The results of an experimental study of the obtained polyvalent vaccine prepared from 7 strains of Pseudomonas aeruginosa showed that after a single immunization of animals followed by infection with homologous strains of Pseudomonas bacillus, a sufficiently high protective effect was observed already on the seventh day after immunization (Table 1). So, for strains PS. aeruga nosa 1314 and 1317 death protection occurs in 100% of cases, for strains B (J 1312 and 1315 from 80% to 100%, for strain No. 1316 to 80% and only for strain No. 1311 does the protective effect appear low (0 -20%). The high protective effect of the vaccine was on the majority of Ps.aerug 1nosa shtafs (f 1313, 1314, 1315, 1316, and 1317) and 21 days after immunization and 8 weeks. With respect to sht1mma PS. A decrease in the protective effect was observed after 21 days and 8 weeks after the vaccine of animals.

Along with a one-time cited and double subcutaneous immunization of animals. After two-time vaccination of mice followed by infection with the same doses of the test cultures of the pyocyanic sticks and according to the same scheme, the protective effect of the vaccine bjol is in general similar to that obtained by riocuie of a single vaccination of animals. There were no significant fluctuations: G percent of surviving animals after infection and only in relation to the strain PS. aeruginosa number 1312 percent

animal protection against death was maintained at a fairly high level, especially 8 weeks after the vaccine. of If, after a single immunization after 8 weeks, the survival rate of the animal experiment was 0–20%, then, after a two-time immunization, with an interval of 7 days, 60–80% of the stomach survived. (Table 2) ..-

In parallel with experiments on

n survival of animals was carried out, radiation was emitted from the formation of agglutinating antibodies, determined by the reaction of agglutination with formalinized strains of Ps.aeruginosa introducing the vaccine. As with

5 double and single immunization agglutinins were determined in vaccinated uninfected animals at various times after immunization (1, 7, 14, 21, 56 days)

0 and in animals that survived infection (1, 2.3 days)

With a single subcutaneous immunization in the sera of the smallest, an increase in the agglutinin titer to the blue5 purulent daddy is observed already by the 7th day after the introduction of the vaccine (in relation to all 7 strains). Another picture is observed in relation to the strain Ps.aeruginosa No. 1312, the growth of agglutinating antibodies to which already occurs,

0 on the first day compared with the stern of natural antibodies in pure unvaccinated animals. On day 14 "after immunization, the highest titer of agglutinins is observed on the strain

5 PS .aerug i nosa No. 1313 (1: 64P) antibody titers to the remaining six strains range from 1: 40-1: 160. The highest levels of specific antibody titers are found on

0 21 days after immunization (the titer of antibodies to PCs Ps .aeruginosa t 1313 and 1317 was 1: 1280, to strain 1311 - 1: 640, to strains No. 1315 and 1316 - 1: 320, with respect to strains No. 1312 and 1314 - did not exceed 1: 160) Further, a decrease in the level of specific antibodies is observed: by 8 weeks in strains No. 1314, 1311, 1315, 1316, they remained at a rather high level (1: 160 - 1: 320),

 whereas for strains nos. 1312, 1313 and 1317, their decrease is observed almost to the initial level (1: 40-1: 20)

With double subcutaneous immunization. The increase in titer agglutinating

5 antibodies were identical, with the only reason that, starting from the first day after the last vaccination, the level of antibodies to all seven strains of Pz, aeruginosa significantly exceeded the level

0 after a single vaccination, i.e. agglutinin titers increased significantly faster and higher levels of antibodies were observed. So, already on the first day after the last immunization, the titer of agglutinins to the Ps strains. ae g ug i nosa (J # 1313, 13127l 1311, 1317 and 1314, 1315 and 1316 amounted to Is 640, 1: 320, 1: 160 and 1:80, respectively. Later, there was an increase in antibody titers and by 21 days the highest level of agglutinins to four strains - №№ 1311, 1312 1313 and 1314 - reached 1: 1280, to the other three 1: 640. By the eighth week after vaccination, there was a slight decrease in the level of antibodies in strains PS .aeruginosa №№ 1311 1314 1315 and 1316, whereas with respect to strains (No. 1312, 1313 and 1317 a decrease in the level of specific antibodies was observed to 1:40. In the study of the titers of agglutinins of immunized animals after infection and on days 1, 2, and 3 it was found that infection of vaccinated mice stimulates an increase in antibody tita on the first day, with their subsequent increase. Thus prepared, the blue-pus multivalet corpuscular vaccine provides good protective immunity and its long-term preservation homologous strains in subcutaneous immunization of animals. The frequency of vaccination is not critical for the manifestation of a protective effect, but has an effect on the level of specific antibodies. . The vaccine can be used for immunoprophylaxis of wound and burn infections, as well as an immunizing drug for the preparation of hyper-hyper-antisex plasma, intended for immunotherapy of patients with pseudomonas sepsis., -, Table 1

30 30 30 30 30

01 02 03 04

05

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Claims (1)

1. Latent Germany No. 1667908, ::. „Kl, 30 h 6, published. 1970.