可生物降解高分子乙型肝炎疫苗微球的免疫原性研究  

Immunogenicity of PELA microsphere encapsulated hepatitis B vaccine

作　　者：廖雪雁[1] 梁争论[1] 周绍兵[2] 张弛[1] 邓先模[2] 张华远[1] 李河民[1] 

机构地区：[1]中国药品生物制品检定所 [2]中国科学院成都有机化学研究所

出　　处：《中华微生物学和免疫学杂志》2002年第6期629-634,共6页Chinese Journal of Microbiology and Immunology

摘　　要：目的　研究聚乳酸 聚乙二醇共聚物 [poly dl lactide poly(ethyleneglycol) ,PELA]包被汉逊酵母表达的HBsAg制备所得乙型肝炎PELA疫苗微球 (PELA·HBsAg)的免疫原性。方法　应用PELA·HBsAg皮下免疫BALB c小鼠 ,在体液免疫方面 ,采用固相放免法测定抗 HBs的水平 ,对抗 HBs中IgG的亚类进行了ELISA测定。在细胞免疫方面 ,于免疫后用3H TdR法测定单个核细胞 (MNC)对ConA、LPS和HBsAg的增殖反应性 ;小鼠迟发型超敏反应 (DTH)的发生程度 ;淋巴细胞亚群分析 ;检测HBsAg特异性CTL活性等评价细胞免疫。结果　①免疫 4周后 ,PELA·HBsAg与HBsAg混合 ,产生抗 HBs的滴度高于铝佐剂的HBsAg[Al(OH) 3 HBsAg]的 2倍多 ;②PELA·HBsAg与HBsAg混合以及PELA·HBsAg产生的IgG2a与IgG的比例显著高于与铝佐剂的HBsAg ;③PELA·HBsAg免疫 2 5d小鼠脾细胞的MNC体外经ConA、LPS刺激后SI最高 ,达到 133.5 9、4 7.4 8,显著高于Al(OH) 3 HBsAg组 (2 8.98、2 1.81)和HBsAg组 (34.95、14 .75 ) ,P <0 .0 5 ;④PELA·HBsAg免疫小鼠后对SRBC所诱导的DTH反应程度有显著的增强作用 (P <0 .0 5 )。⑤PELA·HBsAg与HBsAg混合后免疫小鼠的脾细胞对P815 HBs细胞的特异杀伤率显著高于其它组别 (P <0 .0 5 )。Objective To investigate the immunogenicity of [poly dl lactide poly (ethylene glycol),PELA]encapsulating hepatitis B surface antigen (HBsAg) expressed by Hansenular polymorpha. Methods Encapsulated HBsAg microspheres were administered to BALB/c mice. The anti HBs of PELA microsphere vaccine and HBsAg/microsphere vaccine immunization were detected by solid radioimmunoassay. IgG subclass was titred by ELISA. In the respect of cellular immunity, By using PELA·HBsAg as immunogens, splenic mononuclear cells (MNC) were prepared at 5 , 10 , 15 , 20 and 25 day after immunization respectively. The proliferation of MNC induced by Con A or LPS was assayed. T lymphocyte proliferation tests of the specific antigens and lymphocyte subpopulations were determined. The delayed type hypersensitivity (DTH) determined by footpad swelling using sheep red blood cells (SRBC) as sensitinogen was also tested. Cytotoxic T lymphocyte(CTL )activity was tested by 51 Cr release assay against P815 HBsAg. Results ① The anti HBs titer of HBsAg/microsphere vaccine immunization was 85mIU/ml while that of HBsAg with alum was 36mIU/ml. ② The ratio of IgG2a and IgG in PELA microsphere vaccine and HBsAg/PELA microsphere vaccine was significantly higher than that of HBsAg with alum ( P < 0.05 ). ③ On the 25th day after injection, the SI of Con A or LPS induced proliferation of spleen MNC from mice immunized with PELA·HBsAg was 133.59, 47.48, which was significantly higher than those of Al(OH) 3 HBsAg and HBsAg( P <0.05). ④ PELA·HBsAg significantly improved SRBC induced DTH response of mice ( P < 0.05 ). ⑤ The specific cytolytic activity of splenocyte of mice immunized with PELA·HBsAg microsphere vaccine was significantly higher than that of other groups ( P <0.05). Conclusion PELA had the function to improve the HBsAg induced humoral and cellular immunities,and also played a role in control release of the immune effects of the antigens.

关 键 词：乙型肝炎疫苗 免疫佐剂 聚乳酸-聚乙二醇共聚物 细胞免疫 PELA 固相放射免疫法 

分 类 号：R392[医药卫生—免疫学]