Li Xiaomin, Zhang Yannan, Yang Binyan, Wu Changyou
Institute of Immunology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2015 Nov;31(11):141-1446.
To investigate the secretion of IL-12p40 and IL-6 by splenocytes, dendritic cells stimulated by different Toll-like receptor (TLR) agonists or in the sera of mice immunized with different TLR agonists, and evaluate the effects of different TLR agonists on the function of T helper (Th) cells, especially the differentiation of Th1 cells.
Supernatants of splenocytes and dendritic cells stimulated with different TLR agonists for 24 hours or sera of mice immunized with different TLR agonists at different time points were used to determine the levels of IL-12p40 and IL-6 by ELISA. CD4⁺ T cells isolated from the spleens of ovalbumin-T cell receptor (OVA-TCR) transgenic BALB/c (DO11.10) mice were co-cultured with antigen presenting cells (APCs) from congenic BALB/c mice at 1:3 ratio of T:APCs. Cultures were stimulated with OVA peptide or OVA peptide plus different doses of TLR agonists and the supernatants collected at different time points were assayed by ELISA for detecting IFN-γ.
Pam3CSK4, R848 and CpG oligodeoxynucleotide (ODN) promoted the production of IL-12p40 and IL-6 by splenocytes and dendritic cells obviously, and induced the expression of IFN-γ in antigen specific CD4⁺ T cells in a time- and dose-dependent manner. Monophosphoryl lipid A from Salmonella minnesota R595 lipopolysaccharide (MPLA-SM) induced low levels of cytokines by splenocytes and couldn't promote the production of IFN-γ by antigen specific CD4⁺ T cells, but increased the expressions of IL-12p40 and IL-6 by DCs. All the sera of mice immunized with the four TLR agonists expressed high levels of IL-12p40 and IL-6.
Splenocytes, DCs stimulated or sera of mice immunized with different TLR agonists produced different levels of cytokines, which could further affect the differentiation of Th1 cells.
研究不同Toll样受体（TLR）激动剂刺激的脾细胞、树突状细胞或用不同TLR激动剂免疫的小鼠血清中白细胞介素-12p40（IL-12p40）和白细胞介素-6（IL-6）的分泌情况，并评估不同TLR激动剂对辅助性T（Th）细胞功能，尤其是Th1细胞分化的影响。
用不同TLR激动剂刺激24小时的脾细胞和树突状细胞的培养上清液，或在不同时间点用不同TLR激动剂免疫的小鼠血清，采用酶联免疫吸附测定（ELISA）法测定IL-12p40和IL-6水平。从卵清蛋白-T细胞受体（OVA-TCR）转基因BALB/c（DO11.10）小鼠脾脏中分离的CD4⁺ T细胞，以T细胞与抗原呈递细胞（APC）1:3的比例与同基因BALB/c小鼠的APC共培养。用OVA肽或OVA肽加不同剂量的TLR激动剂刺激培养物，并在不同时间点收集上清液，采用ELISA法检测干扰素-γ（IFN-γ）。
Pam3CSK4、R848和CpG寡脱氧核苷酸（ODN）明显促进脾细胞和树突状细胞产生IL-12p40和IL-6，并以时间和剂量依赖方式诱导抗原特异性CD4⁺ T细胞中IFN-γ的表达。来自明尼苏达沙门氏菌R595脂多糖的单磷酰脂质A（MPLA-SM）诱导脾细胞产生低水平的细胞因子，且不能促进抗原特异性CD4⁺ T细胞产生IFN-γ，但可增加树突状细胞中IL-12p40和IL-6的表达。用这四种TLR激动剂免疫的小鼠所有血清中IL-
