摘要
目的研究rALR对HBsAg及BSA免疫大鼠产生抗体、细胞因子和对脾脏细胞增殖的影响。方法甲醇诱导表达rALR,测定活性并进行以下研究。①rALR对HBsAg免疫的影响:实验分为:生理盐水、HBsAg20μg/只、HBsAg20μg+rALR100μg/kg、HBsAg20μg+rALR25μg/kg、HBsAg20μg+pPIC9K表达上清、HBsAg20μg+CsA10mg/kg,共6组;②rALR对BSA免疫的影响:实验分为:生理盐水、BSA25μg/只、BSA25μg+rALR100μg/kg、BSA25μg+pPIC9K表达上清、BSA25μg+CsA10mg/kg,共5组。以上均皮下注射免疫大鼠,1次/周×4次,ELISA检测血清中相应抗体、IL-2和IFN-γ。③rALR对大鼠脾细胞增殖的影响:Wistar大鼠先皮下注射HBsAg(20μg/只)1次,2周后处死,分离脾单核细胞,种板,再加HBsAg1μg/孔和/或相应处理因素(rALR、空质粒表达产物等),48h后加3H-TdR,12h后收集细胞,检测cpm值。结果rALR100μg/kg+HBsAg组的8只动物中,有2只出现抗HBs的抗体,空质粒对照组和单用HBs Ag组8只动物均出现抗HBs。rALR100μg/kg+BSA组的8只动物中,有3只出现抗BSA抗体,空质粒对照组和单用BSA组8只动物均出现抗BSA的抗体。rALR100μg/kg能明显抑制细胞因子IL-2及IFN-γ的产生。rALR4μg体外能明显抑制脾细胞的增殖。结论rALR能抑制HBsAg和BSA诱导大鼠产生相应抗体及IL-2、IFN-γ的产生;体外能抑制经体内致敏的脾细胞的增殖,说明rALR有免疫抑制作用。
Objective To observe the immunosuppressive effects of rat augmenter of liver regeneration （rALR） in rat immunized by HBsAg and bovine serum albumin （BSA）. Methods rALR was expressed in GS115 under the induction of methanol. The biologic activity of rALR in stimulating proliferation of QGY cell was demonstrated by 3 H-TdR incorporation assay. The rALR was mixed with HBsAg or BSA, respectively, and the intermixture was subcutaneously injected in rats once a week for 4 weeks. One week after last injection, all rats were sacririced and rat serum was collected by centrifugation at 1003 rpm× 5 rain. The ELISA assay was used to detect the anti-HBs, anti-BSA, IL-2, and IFN-γ. Some rats were immunized by subcutaneous injection of HBsAg only one time and sacrificed after two weeks. The splenocytes of the rats were separated and stimulated by HBsAg again in vitro, with addition of different concentrations of rALR. ^3 H-TdR incorporation assay was used to detect the proliferation of splenocytes at 48 h after the stimulation. Results In the HBsAg and the HBsAg （20 μg） ＋ pPIC9K groups, anti-HBs could be detected in all 8 rats; in the saline solution and HBsAg （20 μg） ＋ CsA group, the anti-HBs did not be detected; in the HBsAg （20 μg） ＋ rALR （ 100 μg/kg） group, only 2 of 8 rats had the anti-HBs. Did it be so in the rats immunized with BSA. rALR （ 100 μg/kg） also could inhibit the productions of IL-2 and IFN-γ. RALR also could suppress the proliferation of splenocytes in vitro. Conclusion rALR could inhibit antibody production in rats induced by HBsAg and BSA. rALR also could inhibit the expression of IL-2, IFN-γ and the proliferation of splenocytes in vitro. These data revel that rALR had immunosuppressive effects and could contribute to immunological tolerance on hepatitis B virus infection.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第6期606-609,共4页
Immunological Journal
基金
国家自然科学基金资助项目(30570826)
关键词
肝再生增强因子
外原性抗原
免疫抑制
Augmenter of liver regeneration
Exogenous antigen
Immunosuppression
