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r}HB 3μg,于免疫后第4、7、10、14、25和35 d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10-14 d;rHP诱导CTL活性(P=0.000<0.05)和分泌IFN-γ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莆组(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-HBs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 differentkinds of recombinant hepatitis B(rHB) vaccines in the immunized mice. Methods At serial time points,the levels of IFN-γ 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 FIBsAg MHC class I peptide S28.39 or HBsAg. The lymphocytotoxieity 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-γ and IL-2 in vaccinated mice were detected by ELISPOT, 10-14 days after immunization. The CTL and the level of IFN-γ 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(7dt=4.595,P=0.001<0.05;14dt=5.721,P=0.000<0.05) after immunization. The cellular immune response to the rHP vaccine was thestrongest 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 anti-HBs 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 the three kinds of rHB vaccines were different in their patterns and levels. According to the intensity of early cellular immune response, the two yeast HB vaccines were superior to the rCHO vaccine, especially to the rHP vaccine. In contrast, the rCHO vaccine induced early seroconversion and high levels of anti-HBs.
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