生物可降解微球作为HPV-E7蛋白免疫佐剂的研究

目的：对聚乳酸聚羟基乙酸（PGLA）作为疫苗运输载体进行免疫学评价。方法：用复乳法制备PLGA微球,通过表面吸附人乳头状瘤病毒（HPV）E7蛋白制备成聚乳酸聚羟基乙酸（PGLA）微球,考察粒径分布情况及体外释放水平,通过皮下免疫注射途径免疫C57BL/6小鼠,用间接ELISA法检测免疫鼠血清中的抗体水平,由此评价PLGA微球疫苗运输载体的佐剂效应。。结果：复乳法制备的PLGA微球表面光滑,大小均匀,包封率20.1%,注射小鼠6周后（第2周加强免疫1次）,微球疫苗诱导产生的IgG1抗体水平较同剂量的铝佐剂组和溴化二甲基双十八胺（DDA）组明显升高,（平均滴度分别为3805、1270、2262）;微球疫苗诱导产生IgG2b的抗体水平明显高于铝佐剂组,略低于DDA组,（平均滴度分别为1131、475、2653）而IgG2c的抗体量高于铝佐剂组和DDA组（平均滴度分别为150、36、106）。结论：人乳头状瘤病毒E7蛋白聚乳酸羟基乙酸微球作为疫苗输送体系可以明显的提高抗原的免疫原性。

Research on Biodegradable Microspheres as HPV-E7 Protein Adjuvant

Objective：To evaluate the immune response of PLGA-MS used as carrier to carry the vaccinum.Methods：Microsph-eres were prepared by double——emulsion solvent evaporation method.HPV E7 protein was used an antign absorb different adjuvant conclude aluminum hydroxide hydrate Al（OH）3],Dimethyl dioctadecylammonium（DDA） and poly（lactic-co-glycolic） acid（PLGA） microspheres to immunize C57BL/ 6 mouse with the subcutaneous administration.The IgG1、 IgG2b and IgG2c were detected by indirect ELISA.Result：The microspheres had good shape with smooth and uniform surface,and the mean particle size were about 1 um and surface associated protein were 20.1%.6 weeks later,IgG1 antibody levels in C57BL/ 6 mouse injected by E7 protein of HPV-loaded PLGA microsphere was higher than those injected with E7 protein of HPV puls Al（OH）3 and DDA（the midpointtiters were 3805,1270,2262,respectively）;IgG2b antibody levels was significantly higher than that of Al（OH）3 and slightly less than that of DDA（the midpointtiters were 1131,475,2653 respectively）;In terms of the IgG2b antibody levels,It is higher than that of Al（OH）3 and DDA（the midpointtiters were 150,36,106 respectively）.Conclusion：Biodegradable PLGA microsphere was an effective system for conteolled delivery of vaccines.