CLEC2B Gene Regulates Cell Factors Produced By Lymphocytes And Impacts On Melanocytes

ObjectiveTo investigate the functions of new vitiligo relative gene:CLEC2B(C-type lectin domain family2, member B),for revealing its pathopoiesis target points. To estabish the transient silencing system about lymphocytes and observe the following effect that how lymphocytes’cell factors IL-4mRNA、sIL-2RmRNA、IL-6mRNA levels changed after suppressing the expression of CLEC2B gene in lymphocytes. To detecte whether lymphocytes’ supernatant could infulence melanocytes proliferation or not, and how lympohoctes’ proliferation changed when its CLEC2B gene expression was silenced, thereby represent CLEC2B participation path in the immunity mechanism of vitiligo.MethodsThe lymphocytes lines Jurkat and melanocytes lines B16were seclected. Transfecting corresponding small interfering RNA(siRNA) into lymphocytes, which technique was called RNAi, the suppression effect of CLEC2B gene was detected by real time PCR, also the expressions of IL-4、sIL-2R and IL-6mRNA were detected by real time PCR. The proliferation of melanocytes and lymphocytes were detected by MTT.Results1. The transient silencing systerm about lymphocytes need the compatible proportion between liposomes and siRNA:2μl:80nM, which could silence CLEC2B gene expression in lymphocytes successfully.2. Levels changes of lymphocyte proliferation and cell factors expression after CLEC2B gene silenced:lymphocyte proliferation has no significantly change; IL-4mRNA were up-regulated; sIL-2RmRNA were down-regulated; IL-6mRNA were no significantly influenced. When the lymphocytes’CLEC2B gene was silenced, lymphocytes’ supernatant could decrease the suppression to melanoctyes proliferation, so as to creat beneficial environment for melanoctyes living. ConclusionsCLEC2B gene could exist and express in lymphocytes. Silencing this gene could directly regulate lymphocytes’ cell factors transcription levels, but lymphocytes’ proliferation might not be influenced. CLEC2B could influence melanocytes proliferation by the way of affecting lymphocytes’ functions, all above hinted CLEC2B could accomadate lymphocytes’ actions, then altered the living microenviroments of melanocytes, hence, participated in the immunity mechanism of vitiligo development. The experiment results provided new evidences and views of researching signal transduction path about CLEC2B.