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Jiang J.-J.,Jiangsu Xian Sheng Wei Ke Biological Technology Stock Co. | Pan X.,Jiangsu Xian Sheng Wei Ke Biological Technology Stock Co. | Qian R.,Jiangsu Xian Sheng Wei Ke Biological Technology Stock Co. | Wu Q.-L.,Jiangsu Xian Sheng Wei Ke Biological Technology Stock Co. | And 3 more authors.
Chinese Journal of Biologicals | Year: 2014

Objective To determine the immune activity of tracheitis vaccine by using micro-quantitative hemolysis spectrophotometry. Methods BALB / c mice were divided into one blank control, one negative control and three test groups. The mice in blank control group were unimmunized, while those in negative control group were immunized s.c. with physiological saline, and those in test groups 1, 2 and 3 with three batches of tracheitis vaccine, 0. 5 ml for each, on days 1, 7 and 14. The mice, except those in blank control group, were injected i.p. with 20% sheep red blood cell (SRBC) suspension on day 22 0. 2 ml for each, and killed 4 d later, of which the spleens were prepared into splenocyte suspension. The splenotye suspension was incubated with SRBCs at 37 °C to dissolve the SRBCs around the antibody-forming cells with the participation of complement. The hemoglobin released after dissolution of SRBCs in the hemolytic reaction immediate by antibody-forming cells was measured by microplate reader. Another seventy mice were divided into seven groups, 10 for each. The mice in blank control group were unimmunized, while those in negative control group were immunized with physiological saline, and those in test groups 4, 5, 6, 7 and 8 tracheitis vaccine at concentrations of 6 x 108, 3 x 108, 1. 5 x 108, 0. 75 x 108 and 0. 375 x 108 bacteria/ml respectively. The immune activities of vaccine at various concentrations were determined by above-mentioned method. Results The hemoglobin levels released after dissolution of SRBCs in the hemolytic reaction immediate by antibody-forming cells were significantly higher in three test groups than in negative control group (P < 0. 05). However, the level showed a decreasing tendency with the decreased concentration of tracheitis vaccine. The immune activities of tracheitis vaccine at concentrations of more than 1. 5 X 108 bacteria/ml were significantly higher than (P<0.05), while those at concentrations of less than 0. 75 x 108 bacteria/ml showed no significant difference with, that in negative control group (P> 0. 05). Conclusion Micro-quantitative hemolysis spectrophotometry may be used for the determination of immune activity of tracheitis vaccine. The vaccine at concentrations of more than 1. 5 x 108 bacteria/ml enhanced the immune function of mice. Source

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