Mishra K K, Srivastava A K, Kar S K
Centre for Biotechnology, Jawaharlal Nehru University, New Delhi, India.
Immunol Cell Biol. 1994 Oct;72(5):419-26. doi: 10.1038/icb.1994.62.
We have analysed the suppressive effect of cyclophosphamide, when given in vivo, on the antibody response of BALB/c mice against Shigella dysenteriae antigens using ELISA and immunoblot. Out of various protocols tried, it was found that co-injection of cyclophosphamide at 150 mg/kg bodyweight, i.p., at the time of antigen administration and then after a lapse of 24 h during both primary and secondary immunizations, was the most effective in suppressing antibody response of mice. Analysis of sera by ELISA demonstrated the presence of some antibodies to S. dysenteriae antigens after secondary immunization, but immunoblot analysis using the same sera revealed complete suppression of antibody response. Animals whose antibody response was almost completely suppressed after two immunizations with co-injection of cyclophosphamide, when immunized again after the lapse of 14 days from the date of secondary immunization with Shigella antigens but without administration of cyclophosphamide, partially recovered their ability to respond to the same antigens. This protocol can now be used in mice to analyse the hierarchy of immunogenic epitopes present in a complex mixture of antigens.
我们使用酶联免疫吸附测定（ELISA）和免疫印迹法，分析了环磷酰胺在体内给药时对BALB/c小鼠针对痢疾志贺氏菌抗原的抗体反应的抑制作用。在尝试的各种方案中，发现腹腔注射150mg/kg体重的环磷酰胺，在初次和二次免疫时于抗原给药时及之后24小时注射，对抑制小鼠抗体反应最为有效。ELISA分析血清表明，二次免疫后存在一些针对痢疾志贺氏菌抗原的抗体，但使用相同血清的免疫印迹分析显示抗体反应完全受到抑制。在用环磷酰胺共同注射进行两次免疫后抗体反应几乎完全被抑制的动物，在二次免疫痢疾志贺氏菌抗原后14天，不再注射环磷酰胺再次免疫时，部分恢复了对相同抗原的反应能力。该方案现在可用于小鼠，以分析复杂抗原混合物中存在的免疫原性表位的层次结构。
