伤寒杆菌SOD抗体对小鼠腹腔吞噬细胞杀灭伤寒杆菌的影响

Influence of the anti Salmonella typhi Fe SOD antibody on phagocytic killing effect of mice peritoneal macrophages to Salmonella typhi

为了制备伤寒杆菌ＳＯＤ抗体，研究该抗体对小鼠腹腔吞噬细胞杀灭伤寒杆菌的影响，探讨ＳＯＤ 与伤寒杆菌抗吞噬作用的关系。采用纯化的伤寒杆菌（Ｓｔｗ ｌ）Ｆｅ－ＳＯＤ 免疫家兔制备兔抗Ｆｅ－ＳＯＤ抗体。利用ＥＬＩＳＡ 法检测抗体的效价。免疫电泳、双向琼脂扩散试验和抗体对酶活性抑制试验分析抗体的特性。用不同浓度的抗体预先处理伤寒杆菌，然后进行小鼠腹腔吞噬细胞的吞噬杀菌试验。试验结果抗体的ＥＬＩＳＡ 效价为１∶１０２４０，免疫电泳结果显示纯化的Ｆｅ－ＳＯＤ 及伤寒沙门氏菌无细胞溶解物与Ｆｅ－ＳＯＤ抗体在相同位置各形成一条沉淀线。双向琼脂扩散试验结果显示该抗体与牛红细胞ＳＯＤ 不形成沉淀线，抗体对酶活性抑制试验结果显示抗体可抑制伤寒杆菌及鼠伤寒杆菌无细胞溶解物中部分ＳＯＤ 活性，对牛红细胞ＳＯＤ 的抑制率较低。小鼠腹腔吞噬细胞对伤寒杆菌的吞噬杀菌结果显示经１ ｍ ｌ抗体处理的细菌在６０ ｍ ｉｎ、１２０ ｍ ｉｎ 时和经２ ｍ ｌ抗体处理的细菌在３０ ｍ ｉｎ、６０ ｍ ｉｎ 及１２０ ｍ ｉｎ 时ＣＦＵ值明显低于对照组（Ｐ＜ ０．０１）。伤寒杆菌ＳＯＤ 抗体可促进小鼠腹腔吞噬细胞对伤寒杆菌的杀灭，说明此抗体具有免疫保护作用，也间接地说明了伤寒杆?

s Objective:To prepare the anti Salmonella typhi Fe SOD antibody and study the effect of this antibody on phagocytic killing of mice peritoneal macrophages to S.typhi,thereby clarifing the relationship of the anti phagocytic kill of S.typhi to its SOD.Methods:The anti Fe SOD antibody was rasied in the rabbit immunized with purified S.typhi Fe SOD.The titer of the antibody was determined by ELISA.The properties of the antibody were determined by immuno electrophorisis、double agar difusion and enzym activity inhibited test.Different concentration of the antibody was mixed with the S.typhi,then tested the effect of the antibody on phagocytic killing of mice peritoneal macrophages to S.typhi.Results:The titer of the antibody was 1∶10240.The results of immuno electrophorisis shown that the cell free extracts and the purified Fe SOD from S.typhi formed one precipitated line with the antibody in the same area respectiively.The SOD from bovine erythrocyte did not form precipitated line with the anti Fe SOD antibody.The 42.6%~73.4% of SOD activities in cell free extracts from different Salmonella were inhibited by the antibody in NBT reduction inhibition test,and 24.9% SOD activity from bovine erythrocyte was inhibited by the antibody.The bacteria number of the groups dealing with 1 ml and 2 ml of the antibody were less than that of the control groups at 60 min、120 min and 30 min、60 min、120 min respectively in the phagocytic killing of mice peritoneal phagocytes on S.typhi in vitro(p<0.01).Conclusion:The results shown that the anti Fe SOD antibody can increase the phagocytic killing of mice peritoneal phagocytes on S.typhi in vitro and elucidated that the SOD of S.typhi related to its anti phagocytic kill by peritoneal phagocytes indirectly.