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• W2041555547 abstract "Beni‰ek Z. , J . Sul i , · . ·vraek, J . MojIi‰ova, D. Takaaova, J . Zavadova, R. Ondrejka, A. Ondrejkova: Experimental Inactivated Concentrated Adjuvant Rabies Vaccine. Evaluation of its Efficacy in Cattle. Acta Vet. Brno 2000, 69: 39–44. Efficacy of an experimental inactivated concentrated and purified rabies vaccine, potentiated with lipoid adjuvant was evaluated in comparative experiments performed in cattle by testing of specific cell-mediated immune response (CMI). Commercial inactivated rabies vaccines – Lyscelin (Czech Republic) and Rabisin (France) were used for comparison. Together with CMI evaluation by leukocytes migration inhibition test (LMI), postvaccination rabies antibodies were quantified by the ELISA method and by the rapid fluorescent focus inhibition test (RFFIT) on days 14, 28, 60 and 180 after immunization of the animals. The results of antigenic activity of tested vaccines show the best efficacy of the experimental vaccine from the point of view of humoral immunity, as well as cell-mediated immunity, even though all tested vaccines induced sufficient level of rabies antibodies (≥ 0.5 IU/cm3) at all evaluated intervals. Rabies vaccine, cell-mediated immunity, humoral immunity, cattle In model experiments on laboratory animals efficacy of rabies vaccines can be evaluated on the basis of their immunogenic activity (e.g. NIH method – standard test recommended by WHO 1992; OIE 1996) or antigenic activity – determination of humoral but also cellular immune response after vaccination of target animals. Almost an absolute correlation was proved between the titre of serum rabies antibodies and the level of rabies protection (Sikes et al. 1971; Blancou et al. 1983). Rabies antibodies can be quantified by more than 14 various techniques. At present, classic virus neutralization test on mice – VNT (Atanasiu 1973), rapid fluorescent focus inhibition test – RFFIT (Smith et al. 1973) and ELISA test (Atanasiu et al. 1977) are the most frequently used ones. Standardization of these tests is ensured by using the international standard reference serum with a declared value of international units in 1 cm3 (IU/cm3). Animals vaccinated against rabies must have a minimal level of rabies antibodies 0.5 IU/cm3 one month after immunization (European Pharmacopoeia 1997). Protective effect of rabies vaccination consists not only in the adequate specific antibody response, but it also depends on the defence mechanisms mediated by cells – CMI (Wiktor et al. 1977; Petermann et al. 1976; Murphy 1977). This fact led some authors to the opinion that the test for level determination of specific cell mediated immune response should be used in evaluation of post-vaccination immunity (in evaluation of rabies vaccines efficacy) (Kiss and Tuboly 1981; Pi l le and Matevosyan 1985). Materials and Methods Experiments aimed at comparing and evaluation of the antigenic activity of experimental and commercial rabies vaccines by determination of cellular and humoral immune response on one of the target farm animal species, cattle, were carried out. ACTA VET. BRNO 2000, 69: 39–44 Address for correspondence: MVDr. Zdenek Beni‰ek, CSc. Univerzita veterinarskeho lekarstva Komenskeho 73 041 81 Ko‰ice, Slovenska republika Phone: +421 95 62 29 924 Fax: +421 95 63 23 666 E-mail: benisek@VSVNOV. UVM.sk http://www.vfu.cz/acta-vet/actavet.htm Vaccines In the experiments the following vaccines were used: 1. Experimental – inactivated concentrated purified rabies vaccine (Sul iova et al. 1997; Beni‰ek et al. 1998), potentiated with lipoid adjuvant (Beni‰ek et al. 1999; Sul iova et al. 1999); 2. Commercial inactivated rabies vaccine Lyscelin, op. No. 260594, produced by Bioveta Ltd. Ivanovice na Hane (Czech Republic); 3. Commercial inactivated rabies vaccine Rabisin, op. No. 60H301-0799, produced by Rhone-Merieux (France). Tested animals Young fattened cattle of 250-300 kg were used in the experiments. A total of 25 animals were tested on the basis of non-specific cellular immunity level, determined by a dermatological test with dinitrofluorinebenzene – DNFB (Paul ik and Vrzgula 1989; Paul ik et al. 1991). Fifteen animals with the highest cellular immunity level (with the highest reactive number) were selected into 3 groups and used in the experiments; and the individual groups were averaged. The number of animals was 5 in each group. Immunizat ion scheme Group 1: experimental vaccine with lipoid adjuvant was administered to the animals intramuscularly (i.m.) at a dose of 1.0 cm3. Group 2: vaccine Lyscelin was applied i.m. at a dose of 2.0 cm3. Group 3: vaccine Rabisin was applied i.m. at a dose of 1.0 cm3. Blood sampling from the tested animals was performed on day 14, 28, 60 and 180 after immunization. CMI examinat ion For determination of specific cell-mediated immune response on immunization by rabies vaccines, leukocyte migration inhibition test (LMI) (Bendixen 1977) performed by a capillary method was used in the above mentioned intervals. Purified rabies vaccine strain Vnukovo-32/107 as a specific antigen was used in the LMI test. Its optimal concentration was determined before the experiment. This fact is important in the prevention of inducement of non-specific inhibiton of leukocyte migration by the antigen. Undiluted antigen with migration index MI = 1.04 was applied in the experiment. The migration index was calculated as the quotient of leukocyte migration area in the presence of antigen and the leukocyte migration area without antigen. MI 1.1 probable leukocyte migration stimulated by antigen should be considered. Serological examinat ion Rabies antibodies in the serum of experimental animals were quantified in parallel by two methods: ELISA test was carried out employing a kit developed in our laboratory (Sul iova et al. 1988; Beni‰ek et al. 1989). An antispecies conjugate – peroxidase labelled rabbit antibodies to bovine’s IgG – RAB/IgG Px (Sigma, USA) was used. RFFIT test was carried out by the method of Smith et al. (1973) in modification according to Zavadova et al. (1996). An international standard serum – 30 IU/cm3 (Statens Seruminstitut, Copenhagen, Denmark) was used in both tests. Rabies antibodies values in sera of tested animals were expressed in IU/cm3. Stat is t ical evaluat ion of resul ts Statistical evaluation of CMI results as well as of rabies antibodies quantification by the methods ELISA and RFFIT was carried out by means of the Student’s t-test. The results obtained from the Group 1 immunized by experimental vaccine served as the basis for comparison." @default.
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• W2041555547 title "Experimental Inactivated Concentrated Adjuvant Rabies Vaccine. Evaluation of its Efficacy in Cattle" @default.
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