Effects of immunoglobulin D on expression of IgD receptor and protein tyrosine kinase signaling in human CD4+T cells

Yu-jing WU,Heng-shi CHEN,Wen-sheng CHEN,Jin DONG,Xiao-jie DONG,Xing DAI,Qiong HUANG,Wei WEI

(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine,Hefei 230032,China)

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Effects of immunoglobulin D on expression of IgD receptor and protein tyrosine kinase signaling in human CD4+T cells

Yu-jing WU,Heng-shi CHEN,Wen-sheng CHEN,Jin DONG,Xiao-jie DONG,Xing DAI,Qiong HUANG,Wei WEI

(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine,Hefei 230032,China)

OBJECTIVETo observe whether human CD4+T cells could be activated by immunoglobulin D(IgD)via IgD receptor(IgDR)-Lck.METHODSHuman CD4+T cells were purified from peripheral blood mononuclear cells(PBMCs)with microbeads.The viability of T cells were detected by CCK-8.The binding affinity and expression of IgDR on T cells were detected by flow cytometry.The protein expression of IgDR,Lck and P-Lck were analyzed by western blot.RESULTSIgD could concentrationdependent bind to IgDR on CD4+T cells.The expression of IgDR was increased in response to treatment with IgD in a time-dependent and concentration-dependent manner.Stimulating by IgD resulted in enhanced phosphorylation of Lck compared with that in the medium control sample.The expression of Lck was not changed.As inhibitor of PTK,Herbimycin A or A770041,which combined with IgD could significantly inhibit phosphorylation of Lck(Tyr394).The proliferation promoting effect of IgD was blocked by Herbimycin A or A770041.IgD could stimulate CD4+T cell activation and proliferation through upreg⁃ulating activating tyrosine residue of Lck(Tyr394)phosphorylation.CONCLUSIONThese results demon⁃strate that IgD exaggerates CD4+T cell activities,which may be through promoting Lck phosphorylation.

immunoglobulin D;immunoglobulin D receptor;T cells;Lck

The project supported by National Natural Science Foundation of China(81330081;81673444;81603121);and BSKY(XJ201629 and XJ201630)from Anhui Medical University

Wei WEI,Tel:(0551)65161209,E-mail：wwei@ahmu.edu.cn