CAJ | 학술논문

目的比较不同表达系统来源的乙型肝炎(乙肝)表面抗原(HBsAg)免疫小鼠诱导早期脾淋巴细胞抗原特异性细胞免疫应答的特点,探讨影响乙肝疫苗保护效果的因素.方法 3种HBsAg(汉逊酵母、CHO细胞和血源)分别皮下接种不同组小鼠(BALB/c,H-2d),每只3μg,于免疫后4d分离脾单个核细胞(MNC),经细胞分选仪(MACs)分选后,获得纯度高于90％的CD4+和CD8+T细胞,应用ELISPOT测定MNC、CD4+和CD8+T细胞体外刺激后所产生的细胞因子IFN-γ、IL-2斑点数( SFC).结果细胞分选后,汉逊抗原诱导CD8+T细胞分泌IFN-γ的水平和CD4+T细胞分泌IL-2水平均显著高于CHO抗原组(8/10、2/10,P=0.035;6/10、0/10,P=0.005).汉逊抗原组与血源抗原组CD8+T细胞经体外刺激诱导IFN-y全部阳转(10/10),但分泌水平上汉逊抗原组显著高于血源抗原组(t=2.479,P=0.035)；汉逊抗原组诱导CD4+T细胞IFN-γ阳转率及分泌水平均显著高于血源抗原组(10/10、4/10,P=0.005；t=3.967,P=0.003).结论乙肝抗原免疫小鼠4d即可诱导细胞免疫应答,且汉逊酵母抗原早期诱导抗原特异性IFN-γ、IL-2的能力显著高于CHO和血源抗原,与其临床考核母婴传播阻断HBV保护率显著优于CHO和血源乙肝疫苗相一致,为及时接种乙肝疫苗的必要性和高危新生儿选择接种乙肝疫苗的类型提供依据.
목적 비교불동표체계통래원적을형간염(을간)표면항원(HBsAg)면역소서유도조기비림파세포항원특이성세포면역응답적특점,탐토영향을간역묘보호효과적인소.방법 3충HBsAg(한손효모、CHO세포화혈원)분별피하접충불동조소서(BALB/c,H-2d),매지3μg,우면역후4d분리비단개핵세포(MNC),경세포분선의(MACs)분선후,획득순도고우90％적CD4+화CD8+T세포,응용ELISPOT측정MNC、CD4+화CD8+T세포체외자격후소산생적세포인자IFN-γ、IL-2반점수( SFC).결과 세포분선후,한손항원유도CD8+T세포분비IFN-γ적수평화CD4+T세포분비IL-2수평균현저고우CHO항원조(8/10、2/10,P=0.035;6/10、0/10,P=0.005).한손항원조여혈원항원조CD8+T세포경체외자격유도IFN-y전부양전(10/10),단분비수평상한손항원조현저고우혈원항원조(t=2.479,P=0.035)；한손항원조유도CD4+T세포IFN-γ양전솔급분비수평균현저고우혈원항원조(10/10、4/10,P=0.005；t=3.967,P=0.003).결론 을간항원면역소서4d즉가유도세포면역응답,차한손효모항원조기유도항원특이성IFN-γ、IL-2적능력현저고우CHO화혈원항원,여기림상고핵모영전파조단HBV보호솔현저우우CHO화혈원을간역묘상일치,위급시접충을간역묘적필요성화고위신생인선택접충을간역묘적류형제공의거.
Objective To evaluate the early cellular immune responses to three kinds of hepatitis B surface antigen (HBsAg) in the immunized mice.Methods At day 4,the levels of IFN-γand IL-2 secreted by CD4+ and CD8+T cells which selected from splenic 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 of HBsAg or recombinant hepatitis B surface antigen(rHBsAg).Results After selected by MACs,the purity of CD3+/CD4+ and CD3+/CD8+T cell was more than 90％.The positive rate of IFN-γsecreted by CD4+T cells induced by HBsAg derived from Hansenula polymorpha(rHP) was higher than that of HBsAg derived from CHO cell (rCHO).Levels of IFN-γ secreted by CD8+T cells and IL-2 secreted by CD4+T cells induced by rHP antigen were significantly higher than those of rCHO( P＜0.05 ).Meanwhile,levels of IFN-γsecreted by CD4+T cells and CD8+T cells induced by rHP were also significantly higher than those of plasma HBsAg(pHB) (P＜0.05).Conclusion At day 4,the cellular immune responses induced by HBsAg could be detected.But the immune responses induced by the three kinds of HBsAg are different in levels.According to early cellular immune response intensity,the rHP HBsAg are superior to the rCHO and pHB,in accordance with the high protection rate interrupting the mother-infant transmission immunized by rHP vaccine in clinical trial.It provides scientific basis for necessity of timely birth dose of HB vaccine and kind of HB vaccine for high risk newborn infants vaccinated.