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目的探讨不同的Toll样受体(TLR)激动剂对小鼠脾细胞、树突状细胞(DC)和免疫后白细胞介素12p40(IL-12p40)和IL-6分泌的差异,进一步探讨其对辅助性T(Th)细胞功能的影响尤其是对Th1细胞分化的影响。方法不同剂量的TLR激动剂刺激小鼠脾细胞和DC 24 h或免疫小鼠后收集不同时间点血清,用ELISA检测培养上清液或血清中IL-12p40和IL-6的水平。自CD4~+T细胞受体转基因(DO11.10 OVA-TCR)小鼠脾脏的CD4~+T细胞和与其具有相同背景基因BALB/c小鼠脾脏来源的抗原提呈细胞(APC),将两者按1∶3混合培养,用不同浓度的TLR激动剂联合卵清蛋白(OVA)抗原肽刺激培养,收集不同时间点的上清液,ELISA检测γ干扰素(IFN-γ)的水平。结果 Pam3CSK4、R848和Cp G寡核苷酸(ODN)均能明显促进脾细胞和DC产生IL-12p40和IL-6,以时间和剂量依赖的方式促进抗原特异性的CD4~+T细胞表达IFN-γ;来自明尼苏达沙门菌R595脂多糖的单磷酰脂A(MPLA-SM)诱导脾细胞产生细胞因子水平低、不能促进抗原特异性CD4~+T细胞产生IFN-γ,但可促进DC大量表达IL-12p40和IL-6。四种TLR激动剂免疫小鼠后均能明显促进IL-12p40和IL-6产生。结论不同TLR激动剂作用于脾细胞、DC和免疫小鼠后诱导免疫应答产生细胞因子的能力不同,进一步影响Th1细胞的分化。
Objective To investigate the differences of TLR agonists on the secretion of splenocytes, dendritic cells (DCs) and IL-12p40 (IL-12p40) and IL-6 after immunization in mice. The effect of helper T (Th) cell function is especially influenced by Th1 cell differentiation. Methods Different concentrations of TLR agonist stimulated mouse splenocytes and DCs for 24 h or immunized mice. Serum was collected at different time points. The levels of IL-12p40 and IL-6 in the supernatant or serum were detected by ELISA. CD4 ~ + T cells from the spleen of CD4 ~ + T cell receptor transgenic (DO11.10 OVA-TCR) mice and antigen-presenting cells (APCs) derived from the spleen of BALB / c mice with the same background gene (1: 3). The supernatants were collected at different time points after stimulation with different concentrations of TLR agonist and OVA antigen peptide. The level of IFN-γ was detected by ELISA. Results Both Pam3CSK4, R848 and CpG oligodeoxynucleotides (ODNs) significantly promoted the production of IL-12p40 and IL-6 by spleen cells and DCs, and promoted the antigen-specific CD4 ~ + T cells to express IFN in a time- and dose- -γ; Monophosphoryl lipid A (MPLA-SM) from Minnesota Salmonella R595 lipopolysaccharide induced low splenocyte production of cytokines and did not promote the production of IFN-γ by antigen-specific CD4 ~ + T cells, but promoted the production of DCs Expressed IL-12p40 and IL-6. All four kinds of TLR agonists can obviously promote the production of IL-12p40 and IL-6 after mice were immunized. Conclusion Different TLR agonists act on splenocytes, DCs and immune mice to induce immune responses to produce different cytokines, further affecting the differentiation of Th1 cells.