Kabir S
Department of Biochemistry, State University of Groningen, The Netherlands.
Zentralbl Bakteriol Mikrobiol Hyg A. 1989 Jan;270(3):361-72. doi: 10.1016/s0176-6724(89)80004-5.
Antigens from Vibrio cholerae O1 were analyzed by crossed immunoelectrophoresis (CIE) using sera from immunized rabbits. Thirty different anode-migrating antigens were detected in sonicated antigen preparations of V. cholerae. These antigens were numbered in order to establish a reference precipitation pattern. Antigen no. 30 was identified as the lipopolysaccharide (LPS) antigen, because it reacted with (i) periodic acid-Schiff (PAS) reagent and (ii) the affinity-purified anti-LPS antibodies. Treatment with proteinase K demonstrated that most of the precipitation lines were due to proteins, a part of which were localised at the cell surface. The major outer membrane protein was found to be closely associated with the precipitation line due to the LPS (antigen no. 30). The antigenicity and immunogenicity of V. cholerae cells killed by different methods (merthiolate, heat, phenol, formalin) were examined. As determined by CIE, killing with merthiolate preserved most of the major components of V. cholerae. Heat, phenol and formalin altered the antigenic mosaic of V. cholerae. These results suggested that CIE can be used to analyze several aspects of V. cholerae antigens.
利用免疫兔血清，通过交叉免疫电泳（CIE）分析霍乱弧菌O1抗原。在霍乱弧菌的超声破碎抗原制剂中检测到30种不同的向阳极迁移的抗原。为建立参考沉淀模式，对这些抗原进行了编号。第30号抗原被鉴定为脂多糖（LPS）抗原，因为它与（i）过碘酸希夫（PAS）试剂以及（ii）亲和纯化的抗LPS抗体发生反应。用蛋白酶K处理表明，大多数沉淀线是由蛋白质引起的，其中一部分定位于细胞表面。发现主要外膜蛋白与由LPS（第30号抗原）引起的沉淀线密切相关。检测了用不同方法（硫柳汞、加热、苯酚、福尔马林）杀死的霍乱弧菌细胞的抗原性和免疫原性。通过CIE测定，硫柳汞杀死的霍乱弧菌保留了大部分主要成分。加热、苯酚和福尔马林改变了霍乱弧菌的抗原镶嵌模式。这些结果表明，CIE可用于分析霍乱弧菌抗原的几个方面。
