Electrophoretic Studies on Swine Sera Hyperimmunized with Hog Cholera Virus (HCV)

Abstract

The electrophoretic analysis was applied to sera of 15 normal swine, those of 9 swine hyperimmunized with hog cholera virus (HCV), and those of various stages of the immunization of four swine hyperimmunized with HCV.Methods of electrophoretic analysisThe electrophoretic analysis was carried out with the “HT-B Hitachi Tiselius” apparatus. Electrophoretic conditions were; 9-11MA, 100-110V., 40-50 minutes, and 15-22°C. The phosphate buffer solution was used. Total serum protein was determined with the “Hitachi Hand Protein Refractometer”. Protein fraction was measured with a ascending pattern by the weight method.Experimental results1) The results of electrophoretic analysis on sera of 15 swine which were considered to be in normal conditions are shown in Table 1; total protein 7.1g/dl, albumin (alb) 46.2%, α-globulin (glob) 19.7%, β-glob 15.7%, γ-glob 18.4%, and A/G 0.877.2) The results of electrophoretic analysis on antisera of 9 swine hyperimmunized with HCV are shown in Table 2; total protein 7.6g/dl, alb 40.8%, α-glob 19.4%, β-glob 14.7%, γ-glob 25.1%, and A/G 0.701.3) Mathews et al. have reported the increase of γ-glob of sera of swine hyperimmunized with HCV. This paper shows that as they reported, the average percent of γ-glob of hyperimmune sera was observed to be higher than that of normal sera, and that the increase of γ-glob was not always observed in all hyperimmune sera (see Table 2).4) The results of electrophoretic analysis on sera of various stages of the immunization of 4 swine hyperimmunized with HCV are shown in Table 3. No remarkable electrophoretic changes of the sera were observed at any stages of the immunization of the cases concerned, and the antiserum which showed a remarkable increase of γ-glob was not observed in any cases.5) The relationship of viral antibody to the protein fraction of swine sera hyperimmunized with HCV is of considerable importance. This paper reports that the relationship was investigated electrophoretically and immunologically.The serum protein fractions were isolated from hyperimmune sera by means of the starch-electrophoresis and the neutralizing antibody titer of each fraction (alb, α-, β-, γ-glob) was detected by the tissue culture method, and the protective antibody was proved by injection of each fraction into swine.From the above experimental results, it has been found that the neutralizing antibody is contained in β-glob and γ-glob fraction of the hyperimmune sera, and that the protective antibody consists mainly in γ-glob fraction of the hyperimmune sera.