摘要
目的探讨重组日本血吸虫谷胱甘肽-S-转移酶(rSjGST)可生物降解缓释微球对小鼠的免疫效应。方法构建表达质粒pET28a(+)-SjGST,在E.coliBL21内表达日本血吸虫谷胱甘肽-S-转移酶融合蛋白,纯化后以聚乳酸/乙醇酸(PLGA75/25)为包裹材料,制备rSjGST可生物降解性微球。按一定剂量皮下注射免疫微球免疫小鼠,分别于免疫后6、9、12、15、18、21周眼眶采血并取出脾脏,检测血清IgG及脾T淋巴细胞A值,比较各组免疫效果。实验另设FCA佐剂组、铝佐剂组和生理盐水对照组。结果与生理盐水对照组相比,FCA组、铝佐剂组血清总IgG水平在第9周显著升高(P<0.01),且两组均在免疫9周时达峰值,之后逐渐下降,而微球组在21周时仍呈上升趋势(P<0.01);与FCA组、铝佐剂组比较,微球组12周后血清总IgG水平显著升高(P<0.01)。在免疫后6～15周,三免疫组小鼠脾T淋巴细胞A值差异无统计学意义(P>0.05),但在免疫18周后微球组脾T淋巴细胞A值显著高于其他两组(P<0.01)。结论rSjGST可生物降解微球免疫小鼠,具有长效和高效作用。这为血吸虫病疫苗的进一步研究提供了新的思路。
Objective To detect the immune effect of biodegradable microspheres of recombinant glutathione-S-transferase from Schistosoma japonicum （rSjGST） in mice. Methods Expression plasmid pET28a （＋）-SjGST was constructed. rSjGST was expressed by E. coli BL21 and purified. The biodegradable microspheres of rSjGST were prepared with poly lactic-co-glycolic acid （PLGA, 75/25）. Six-week BALB/c female mice of microspheres group were immunized subcutaneously with a single vaccine, and the blood was collected from eyes on the 6th, 9th, 12th, 15th, 18th, 21st week respectively while the spleens were extracted. The NS control group, FCA group and Al-adjuvant group were also built. The total antibody IgG in sera was detected. And absorbance （A value ） of spleen T lymphocyte cells was determined. Results Compared with the NS control group, the levels of total antibody IgG in FCA group and Al-adjuvant group were significantly increased （P〈0.01） and reached the peak at 9th week, and then decreased gradually, while the level of total antibody IgG in the microspheres group still increased at 21st week （P〈0.01）. There was a distinct difference between total antibody IgG of the microspheres group and that of FCA group as well as the microspheres group and Al-adjuvant group （P〈0.01） after the 12th week. There was no difference for spleen cells A values among the three immune groups （P〉0.05） from the 6th week to the 15th week. And there was a distinct difference between A value of the microspheres group and that of FCA group as well as the microspheres group and Al-adjuvant group （P〈0.01） after the 18th week. Conclusion There is significantly long and high effect of biodegradable microspheres of rSjGST on the infected mice, which provides a new means to further study vaccine for S. japonicum.
出处
《中国病原生物学杂志》
CSCD
2007年第4期277-280,共4页
Journal of Pathogen Biology
基金
安徽省教育厅自然科研基金资助项目(No2005kj303)
关键词
日本血吸虫谷胱甘肽-S-转移酶
聚乳酸/乙醇酸微球
疫苗缓释剂
小鼠
Recombinant glutathione-S-transferase from Schistosoma japonicum （rSjGST）
PLGA microsphere
controlled-releasing vaccine
mice
