iNKT细胞联合卵清蛋白对髓源性树突状细胞表型和功能的影响

Effects of iNKT cells from OVA-induced asthmatic mice combined with OVA on phenotype and function of bone marrow-derived dendritic cells in vitro

目的:观察哮喘小鼠恒定自然杀伤T细胞(iNKT细胞)联合卵清蛋白(OVA)对髓源性树突状细胞(BMDCs)表型和功能的影响。方法:取健康雄性野生型BLAB/c小鼠BMDCs,分别与哮喘小鼠脾脏iNKT细胞联合OVA(iNKT细胞联合OVA组)、脂多糖(LPS组)、iNKT细胞联合PBS(iNKT细胞联合PBS组)、OVA(OVA组)或PBS(PBS组)共培养20 h,采用流式细胞术检测各组BMDCs表面分子MHC-II、CD80、CD86和CD40的表达水平,ELISA法检测培养上清液中白细胞介素12(IL-12)p70、IL-6、肿瘤坏死因子α(TNF-α)和IL-10的水平;取各组共培养后的BMDCs,与DO11.10转基因小鼠脾CD4^+ T细胞共培养48 h,采用ELISA法检测培养上清液中IL-4和干扰素γ(IFN-γ)的水平。结果:iNKT细胞联合OVA组BMDCs表面分子MHC-II、CD80、CD86和CD40及培养上清液IL-12 p70、IL-6和TNF-α水平与LPS组比较差异无统计学显著性(P>0.05),但明显高于iNKT细胞联合PBS组、OVA组和PBS组(P<0.05或P<0.01);iNKT细胞联合OVA组BMDCs与DO11.10 CD4^+ T细胞共培养上清液IL-4水平与LPS组比较差异无统计学显著性(P>0.05),但明显高于iNKT细胞联合PBS组、OVA组和PBS组(P<0.01)。结论:存在OVA时,iNKT细胞可以诱导BMDCs表型和功能的免疫原性成熟。

AIM: To investigate the effects of invariant natural killer T-cells(iNKT cells) from ovalbumin(OVA)-induced asthmatic mice combined with OVA on the phenotypic and functional characteristics of bone marrow-derived dendritic cells(BMDCs) in vitro. METHODS: The BMDCs from wild-type(WT) BLAB/c mice were co-cultured with purified iNKT cells from WT mice immunized and challenged with OVA in the presence of 100 mg/L OVA(iNKT cells plus OVA group) or PBS(iNKT cells plus PBS group) for 20 h, and were also cultured with 50 mg/L LPS(LPS group), 100 mg/L OVA(OVA group) or PBS(PBS group) for 20 h. The expression of MHC-II, CD40, CD86, and CD80 on the BMDCs was measured by flow cytometric analysis, and the levels of interleukin-12(IL-12) p70, IL-6, tumor necrosis factor-α(TNF-α) and IL-10 in the culture supernatant were measured by ELISA. Splenic CD4^+ T cells from DO11.10 transgenic mice were co-cultured for 48 h with the above mentioned BMDCs, and then the concentrations of IL-4 and interferon-γ(IFN-γ) in culture supernatants were measured by ELISA. RESULTS: The expression of MHC-II, CD80, CD86 and CD40, and releases of proinflammatory cytokines by BMDCs in iNKT cells plus OVA group were comparable to those in the LPS group(P>0.05), but significantly higher than those in iNKT cells plus PBS group, OVA group, and PBS group(P<0.05 or P<0.01). The concentration of IL-4 in culture supernatants from BMDCs in iNKT cells plus OVA group co-cultured with DO11.10 CD4^+ T cells was similar to that in LPS group(P>0.05), but markedly higher than that in iNKT cells plus PBS group, OVA group, and PBS group(P<0.01). CONCLUSION: Bone marrow-derived dendritic cells undergo immunogenic maturation upon interaction with iNKT cells in the presence of OVA.