鼠伤寒杆菌外膜蛋白作为保护性疫苗的初步实验研究

STUDY OF PRIMARY EXPERIMENT OF OUTER MEMBRANE PROTEIN OF SALMONELLA TYPHIMURIUM AS A PROTECTIVE VACCINE IN MICE

本文采用超声波破碎和Triton X-100处理、超速离心的方法,提取了鼠伤寒杆菌的外膜蛋白(OMP)。用提取的OMP免疫家兔和小鼠,经用ELISA方法测定小鼠、家兔及鼠伤寒意染患者血清中抗-OMP抗体含量,并用小鼠作足垫肿胀实验及主动和血清被动保护试验。结果免疫动物及患者血清中都含有较高滴度抗体,免疫小鼠足垫出现明显地DTH反应。50ug OMP免疫小鼠可保护500 LD50毒株的攻击,0.2ml免疫血清亦能够被动保护以上同样的毒株攻击。这些结果表明,提取的OMP有较强的免疫原性和明显的免疫保护作用,应用Western blot分析免疫血清均能识别36KD蛋白带,36KD蛋白带可能是鼠伤寒杆菌的主要免疫原。

OMP of S.typhinurium Were extracted with the method of described by Schanitman. Ribbits and mice were immunized with the extracted OMP. The serum antibody titer of ribbits, mice arid patients was measured and footpad swelling experiment be done with mice. Mice were used as active and passive protective experiment. The result showed that both ribbits and mice had high antibody titer in serum. Patients infected by S. typhimurium also had high antibody titer in serum. Immunized mice pad exhibited obvious DTH response. Mice immunized with 50ug of OMP could be protected from the attack by 500 LD50 of wild strain of S. typhimurium, and 0.2ml immune serum also could passive protected mice to attacked by above same 500 LD50. This result indicatec that the extracted OMP had strong immunogenicity and obvious immune protection. Immune of OMP was analyed with western blot. The result showed that 36KD OMP band could be indentified by all immune serum of OMP. The 36KD protins (porin) may be a major immunogen of S. typhimurium.