鼠伤寒沙门氏菌外膜蛋白诱发BALB／C小鼠免疫保护的研究

STUDY ON PROTECTIVE IMMUNITY INDUCED BY OUTER MEMBRANE PROTEINS OF SALMONELLA TYPHIMURIUM IN BALB/C MICE

本文采用超声破碎ＴｒｉｔｏｎＸ—１００和超速离心技术提取了鼠伤寒杆菌（Ｓａｌｍｏｎｅｌｌａｔｙｐｈｉｍｕｒｉｕｍ，ＳＴＭ）的外膜蛋白（Ｏｕｔｅｒｍｅｍｂｒａｎｅｐｒｏｔｅｉｎｓ，ＯＭＰｓ），并发现ＯＭＰｓ中脂多糖ＬＰＳ的含量约为５％，ＯＭＰｓ经ＳＤＳ—ＰＡＧＥ显示１０余条蛋白带。对ＯＭＰｓ诱发ＢＡＬＢ／Ｃ小鼠产生典型迟发型变态反应ＤＴＨ和ＩＬ—２的水平进行了检测。经腹腔免疫的小鼠用５００ＬＤ５０鼠伤寒杆菌（５０１１５）攻击，１００％可得到保护；用５００ＬＤ５０伤寒杆菌（Ｅ６８６）攻击，３３．３％可得到交叉保护。免疫ＢＡＬＢ／Ｃ小鼠的Ｔ淋巴细胞，经尾静脉注射给非免疫小鼠，可使后者得到８５．７％的被动免疫保护，上述结果说明ＯＭＰｓ能诱发ＢＡＬＢ／Ｃ小鼠细胞免疫和保护性免疫，并提示成为分子疫苗的可能性。

Outer membrane proteins(OMPs)from Salmonella typhimurium were extracted by using super sonication, Triton X-100 and super speed centrifugation,and in OMPs were found 5% lipopolyaccharide(LPS)approximately.OMPs were shown 10 more bands by SDS-PAGE.In OMPs immunized BALB/C mice significant level of delayed-type hypersensitivity(DTH)response and interleukin-2(IL-2)production were detected.Active immunization with OMPs induced 100%protection to a intraperitoneal(i·P)challenge with 500 LD50 of Salmonella typhimurium(50115)in BALB/C mice, In addition,33.3% crossed protection against challenge with up to 500 LD50 of Salmonella typhi(E686)was also achieved. Furthermore,We found that T cells from the OMPs-immmunized mice could transfer the protection(85.7%)against Salmonella typhimurium to nonimmunity mice.All these results suggest that STM-OMPs can induce more powerful cell-mediated immunity and protective immunity,develop the probability of a molecular vaccine.