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作者用不引起动物全身感染的缺失鞭毛的伯氏疏螺旋体减毒突变株作为疫苗,在小鼠模型中评价其免疫原性和保护效力.作者以伯氏疏螺旋体HB19株及其无鞭毛突株HB19FLa~-和有感染性的低传代株Sh.2和N40制备对数生长晚期的活的或加热灭活的螺旋体悬液,经磷酸缓冲盐水(PBS)稀释后分别于尾根背部及腹部皮肤皮内免疫小鼠(100μl),对照小鼠则注射100μlBSKⅡ培养基PBS稀释液.末次采血后4天用10~4疏螺旋体尾根部皮内接种攻击小鼠,14天后杀死小鼠,以BSKⅡ培养基培养小鼠的血浆、膀胱、心脏及关节标本,并用相差显微镜镜检,如无具有动力的螺旋体即为培养阴性.采用ELISA和生长抑制法检测小鼠免疫应答.通过聚丙烯酰胺凝胶电泳和蛋白印迹法检测鞭毛蛋白和外膜蛋白.
The authors evaluated the immunogenicity and protective efficacy of the attenuated mutant strain of Borrelia burgdorferi that lacked the flagella that caused systemic infection in mice in a mouse model.With Borrelia burgdorferi HB19 strain and its flagella-free strain HB19FLa ~ and infectious low-passage strains Sh.2 and N40 were prepared live or heat-inactivated helicobacter suspension in logarithmic growth phase, diluted in phosphate buffered saline (PBS), respectively, in the tail root and abdomen skin (100 μl), and control mice were injected with 100 μl BSKⅡ medium PBS Diluent Four days after the final blood draw, the mice were inoculated intradermally with 10 to 4 Borrelia caudalis and mice were killed 14 days later and cultured in BSKII The mouse plasma, bladder, heart and joint specimens were cultured on the basis of phase contrast microscopy, negative if no motile spirochete, and the immune response of mice was detected by ELISA and growth inhibition.After polyacrylamide gel electrophoresis The flagellin and outer membrane proteins were detected by Western blot.