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目的探讨日本血吸虫极低密度脂蛋白结合蛋白(SVLBP)重组抗原的免疫保护性及其作为候选疫苗的潜在价值。方法用异丙基-β-D-硫代半乳糖苷(IPTG)诱导克隆菌大量表达SVLBP重组抗原,以镍-次氮基三乙酸琼脂糖树脂(Ni-NTA)亲和层析法纯化制备SVLBP重组抗原;将纯化的重组抗原加福氏佐剂经皮下免疫C57BL/6小鼠,每隔2周免疫1次,在第3次免疫后10d,经腹部皮肤攻击感染日本血吸虫尾蚴35±1条,感染后45d剖杀,计数检获虫数和每克肝虫卵数(LEPG)。小鼠在免疫前和攻击感染后分别采眶窦血,用ELISA测定特异性IgG及其亚群抗体水平。结果相对于佐剂对照组,免疫组小鼠的减虫率为33.4%,减卵率为47.6%;免疫后小鼠产生高水平的特异性IgG(>1:6400);抗体亚类检测结果显示,免疫组小鼠IgG2a、IgG2b、IgG1明显高于免疫前和佐剂对照组。结论日本血吸虫皮层蛋白SVLBP重组抗原能诱导小鼠产生一定的保护性免疫,是潜在的疫苗候选抗原分子。
Objective To investigate the immunoprotection of recombinant antigen of Very Low Density Lipoprotein-Binding Protein (SVLBP) of Schistosoma japonicum and its potential value as a candidate vaccine. Methods SVLBP recombinant antigens were induced by IPTG in large quantities and purified by Ni-NTA affinity chromatography SVLBP recombinant antigen; C57BL / 6 mice were subcutaneously immunized with the purified recombinant antigen plus Freund’s adjuvant, immunized once every 2 weeks. At the 10th day after the third immunization, 35 ± 1 of cercariae of Schistosoma japonicum were challenged by abdominal skin , Cut off 45 days after infection and count the number of seized pests and the number of eggs per gram of liver (LEPG). The mice were infused with orbital sinus blood prior to and after challenge infection, and the levels of specific IgG and its subgroup antibodies were measured by ELISA. Results Compared with the adjuvant control group, the mice in the immunized group had a worm reduction rate of 33.4% and an egg reduction rate of 47.6%. The immunized mice produced high levels of specific IgG (> 1: 6400) The results showed that IgG2a, IgG2b and IgG1 in immunized mice were significantly higher than those in pre-immune and adjuvant control groups. Conclusion The SVLBP recombinant antigen of Schistosoma japonicum cortical protein can induce some protective immunity in mice, which is a potential vaccine antigen candidate.