类风湿关节炎中IgD通过T细胞刺激B细胞活化  

IgD activates B cell function through T cell in RA patients

作　　者：胡晓曦 张爱君 张静 吴育晶[1] 魏伟[1] Hu Xiaoxi;Zhang Aijun;Zhang Jing(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-infammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine,Hefei 230032)

机构地区：[1]安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,抗炎免疫药物安徽省协同创新中心,合肥230032

出　　处：《安徽医科大学学报》2020年第4期572-578,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81603121、81673444、81973332);安徽医科大学博士科研资助基金(编号:XJ201629)。

摘　　要：目的研究免疫球蛋白D(IgD)通过人T细胞上的IgD受体(IgDR)-淋巴细胞特异性酪氨酸激酶(Lck)信号活化T细胞的功能以及对B细胞活化的影响。方法收集健康对照者和类风湿关节炎(RA)患者的外周血,通过流式分选技术分选出外周血单个核细胞(PBMCs)中的CD4^+T细胞和CD19^+B细胞,将不同浓度IgD刺激的CD4^+T细胞与CD19^+B细胞利用transwell小室进行共培养后,激光共聚焦显微镜观察CD4^+T细胞上Lck与IgDR的共定位,Western blot法检测CD4^+T细胞上P-Lck和CD40L蛋白的表达、CD19^+B细胞上CD40蛋白的表达,CCK-8法检测IgD刺激的T细胞对B细胞活化的影响。结果IgD可显著上调健康对照者CD4^+T细胞上Lck与IgDR两种蛋白的共表达及P-Lck蛋白的表达;在健康对照者和RA患者中,IgD刺激的T细胞可诱导B细胞的活化;IgD可显著上调CD40L蛋白在CD4^+T细胞上的表达以及CD40蛋白在CD19^+B细胞上的表达。结论IgD可通过IgDR-Lck信号活化T细胞,并通过上调T-B共刺激分子CD40L和CD40的蛋白表达刺激B细胞的活化。Objective To investigate immunoglobulin D(IgD) can activate the function of T cells through immunoglobulin D receptor(IgDR)-lymphocyte-specific protein tyrosine kinase(Lck) signaling on human T cells and its effect on B cell activation.Methods Peripheral blood from healthy controls and RA patients were collected and peripheral blood mononuclear cells(PBMCs) were separated. CD4^+T cells and CD19^+B cells were sorted by flow cell sorting. Different concentration of IgD-stimul-ated CD4^+T cells were co-cultured with CD19^+B cells in transwell chamber. Laser confocal microscopy was used to observe Lck and IgDR co-localization on CD4^+T cells.Western blot was used to detect the protein expression of P-Lck and CD40 L on CD4^+T cells and the protein expression of CD40 on CD19^+B cells. CCK-8 method was used to detect the effect of IgD-stimulated T cells on B cell activation. Results IgD significantly up-regulated the co-expression of Lck and IgDR proteins and the expression of P-Lck protein on CD4^+T cells of healthy control. Both in healthy controls and RA patients,IgD-stimulated T cells could induce B cell activation. IgD could significantly up-regulate the expression of CD40 L protein on CD4^+T cells and the expression of CD40 protein on CD19^+B cells. Conclusion IgD could activate T cells via the IgDR-Lck signaling and stimulate B cell activation by up-regulating protein expression of T-B co-stimulatory molecules CD40 L and CD40.

关 键 词：免疫球蛋白D CD4^+T细胞 CD19^+B细胞 共培养 类风湿性关节 

分 类 号：R593.22[医药卫生—内科学]