摘要
目的选择目前常用的几种商业化Toll样受体(TLR)配体进行对比,考察各候选物在小鼠体内外诱导增强Th1型细胞免疫反应的特点,为新疫苗佐剂的研究和筛选提供支持。方法TLR配体多肌胞苷酸[poly(I∶C)]、单磷酰脂质A(MPLA)、瑞喹莫德(R848)、含CpG序列寡核苷酸C(CpG-C)体外刺激小鼠树突状细胞(DC),采用ELISA检测细胞上清液白细胞介素12(IL-12)的水平;以卵清蛋白(OVA)为抗原,各TLR配体为佐剂免疫小鼠后,流式细胞术检测引流淋巴结中DC、自然杀伤(NK)细胞比例和表型变化以及CD4^+T细胞、CD8^+T效应细胞比例的变化;ELISA检测免疫小鼠血清中抗OVA抗体IgG2a的浓度。结果在体外实验中,仅CpG-C和R848能显著增强IL-12的分泌。在体内实验中,与其他TLR配体佐剂相比,R848更有效地促进DC和NK细胞向小鼠淋巴结中聚集并诱导CD4^+和CD8^+效应T细胞生成,以及刺激产生大量抗OVA特异性IgG2a。结论与其他免疫佐剂相比,R848更具增强Th1细胞免疫作用。
Objective To compare and characterize the Th1 immune responses induced by the most commonly used commercial Toll-like receptor（TLR） agonists through in vivo and ex vivo experiments.Methods The concentrations of IL-12 were tested by ELISA after mouse dendritic cells（DCs） in vitro were stimulated by one of tested TLR agonists,including poly（I∶ C）,monophosphoryl lipid A（MPLA）,resiquimod（R848）,cytosine polyguanine-C（Cp G-C）.The changes in percentage and phenotype of DCs,NK cells and effector T cells in the draining lymph nodes were analyzed by flow cytometry after BALB/c mice were immunized with ovalbumin（OVA） mixed with selected TLR agonist.The serum concentrations of specific anti-OVA IgG2 a in the immunized mice were determined by ELISA.Results Only Cp G-C and R848 could significantly induce the production of IL-12 from bone marrow-derived DCs in vitro.Among the tested TLR agonists,R848 was the most effective adjuvant in recruiting DCs and NK cells into lymph nodes,inducing the proliferation of CD4^＋ and CD8^＋ effector T cells and the production of specific anti-OVA IgG2 a in vivo.Conclusion R848 was the most potential Th1-promoting adjuvant among the tested TLR agonists.
作者
沈沅
曾梦华
邱峰
唐华
SHEN Yuan ZENG Menghua QIU Feng TANG Hua(Department of Pharmacy Department of Gastrointestinal Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2017年第5期591-596,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(31400791)
