不同种类乙型肝炎重组疫苗诱导免疫应答特点比较  

Comparison of the kinesis of immune responses in mice vaccinated by different kinds of recombinant hepatitis B vaccines

作　　者：胡忠玉[1] 何鹏[1] 方鑫[1] 邱少辉[1] 梁争论[1] 李河民[1] 庄辉[2] 

机构地区：[1]中国药品生物制品检定所,北京100050 [2]北京大学医学部

出　　处：《中华流行病学杂志》2008年第8期810-814,共5页Chinese Journal of Epidemiology

基　　金：国家“十五”科技攻关课题资助项目（2004BA718802）

摘　　要：目的比较三种乙型肝炎重组疫苗（rHB）[乙型肝炎汉逊酵母重组疫苗（rHP）、酿酒酵母重组疫苗（rSC）和CHO重组疫苗（rCHO）]诱导小鼠细胞和体液免疫应答特点。方法给3组小鼠（BALB／c，H-2d）分别皮下接种3种rHB3弘g，于免疫后第4、7、10、14、25和35d分离脾单个核细胞（MNC），应用酶联斑点法（ELISPOT）测定MNCs体外刺激后所产生的细胞因子IFN-γ、IL-2斑点形成细胞数（SFC）。刺激物为多肽或HBsAg。同时测定免疫后不同时间的细胞毒T淋巴细胞反应（CTL）活性和小鼠血清抗-HBs水平。结果应用ELISPOT法，不同种类rHB免疫小鼠后诱导分泌IFN-γ和IL-2峰值均为10—14d；rHP诱导CTL活性（P=0．000〈0．05）和分泌IFN，7水平（P〈0．05）均显著高于rCHO和rSC，且rHP于免疫后第10天诱导CTL杀伤率最高（39．8％）；rHP组诱导分泌IL-2水平均显著高于其他两种疫苗组（P〈0．05），第7和14天时，rSC组诱导产生IL-2水平均显著高于rCHO组（第7天t=4．595，P=0．001〈0．05；第14天t=5．721，P=0．000〈0、05）；免疫后第7天3种疫苗诱导细胞免疫应答强度依次为rHP最强，rSC其次，rCHO最弱；小鼠血清抗-Ugs阳转率和抗体滴度随免疫时间而逐渐上升，第7天时rCHO组抗体显著高于rHP组（P=0．044〈0．05），第14天时rCHO组抗体显著高于rHP组（P=0．009〈0．05）和rSC组（P=0．012〈0．05）。结论3种疫苗免疫小鼠的应答特点不同，酵母重组疫苗早期诱导细胞免疫应答优于rCHO；rCHO的优势是诱导产生抗-HBs早，且滴度较高。Objective To evaluate the kinesis of cellular and humoral immune responses to different kinds of recombinant hepatitis B（rHB） vaccines in the immunized mice. Methods At serial time points, the levels of IFN-y and IL-2 secreted by spleeny mononuclear cells（MNC） of the vaccinated mice were detected by enzyme-linked immunospot methods（ELISPOT） after stimulation in vitro with HBsAg MHC class I peptide S28-39 or HBsAg. The lymphocytotoxicity of the immunized mice were also detected （CTL） by a specific lysis assay and the levels of anti-HBs were measured by the Abbott IMX kit. Results The peak values of IFN-y and IL-2 in vaccinated mice were detected by ELISPOT, 10-14 days after immunization. The CTL and the level of IFN-y induced by rHB vaccine derived from yeast cells（Hansenular polymorpha） （rHP vaccine） were significantly higher than the other two vaccines （P〈 0.05）. The maximum lysis of CTL appeared in the vaccinated mice on day 10 after immunization,with the percentage of 39.8 %. The levels of IL-2 induced by rHP vaccine were significantly higher than the other two vaccines （P 〈 0.05）. However, the IL-2 levels in the rSC （saccharomyces cerevisiae） vaccine group were higher as compared with the rCHO vaccine group at day 7 and day 14（7 d t = 4. 595, P ： 0.001〈 0.05; 14 d t ： 5.721 ,P = 0.000〈 0.05） after immunization. The cellular immune response to the rHP vaccine was the strongest while it was the lowest to the rCHO vaccine at day 7 after immunization. The sero-positive rates and the titers of anti-HBs in the vaccinated mice increased with time after vaccination. The titers of antiHBs in the rCHO vaccine group at day 7 were similar to the rSC vaccine group, but significantly higher than that of the rHP vaccine group（ P = 0. 044 〈 0.05）. The anti-HBs titers of the rCHO vaccine group at day 14 were significantly higher as compared to the rSC（ P = 0.012 〈 0.05） and rHP（ P = 0. 009 〈 0.05 ） vaccine groups. Conclusion The immune responses induced by

关 键 词：乙型肝炎 疫苗 细胞因子 酶联免疫斑点法 斑点形成细胞 细胞毒T淋巴细胞反应 

分 类 号：R392[医药卫生—免疫学]