重组毕赤酵母HBsAg诱导小鼠细胞免疫应答的研究

目的   研究毕赤酵母表达的重组HBsAg(Pichia-rHBsAg)在小鼠中诱导T细胞免疫应答的能力,评价疫苗的免疫原性.   方法  以rHBsAg免疫雌性BALB/c小鼠,制备小鼠脾淋巴细胞,并在体外以抗原/特异性多肽刺激.采用ELISA法测定抗原特异性T淋巴细胞分泌的细胞因子,酶联免疫斑点法(ELISPOT)测定CTL频数.   结果 由毕赤酵母表达的rHBsAg可在小鼠中诱导Th1和Th2型免疫应答;加铝佐剂的rHBsAg诱导IFN-γ的水平及CTL克隆明显高于无佐剂抗原;Pichia-rHBsAg刺激小鼠细胞免疫应答的水平强于酵母-rHBsAg,与中国仓鼠卵巢细胞-rHBsAg相仿.   结论   Pichia-rHBsAg可在BALB/c小鼠中诱导较强的细胞免疫应答.

Induction of cell-mediated immune response in BALB/c mice immunized with recombinant HBsAg expressed in Pichia pastoris

Objective To study the response of T cell-mediated immunity (CMI) induced with HBsAg expressed in Pichia pastoris (Pichia-rHBsAg) in mice and evaluate the immunogenicity of a vaccine. Methods BALB/c mice were immunized with rHBsAg with or without an adj uvant.Then spleenlymphocytes were isolated and stimulated by antigen/specific peptide for investigating CMI response.Cytokines and CTL frequency were detected by ELISA and ELISPOT.respectively. Results PichiarHBsAg could trigger Th1 and Th2 type immune response.Comparing with rHBsAg alone,higher concentration of IFN-γ and more CTL clones were observed with rHBsAg plus AI(0H)3 aajuvant.In addition,the CMI response elicited by Pichia-rHBsAg is stronger than that induced by yeast-rHBsAg,and comparable to that induced by CHO-rHBsAg. Conclusion Pichia-rHBsAg could induce stronger CMI response in BALB/c mice.