Research On Protective Mechanism Of CD30L/CD30 Signaling In The Pathogenesis Of Psoriatic Skin Inflammation Mediated By ?? T17 Cells

Objective:Psoriasis is a skin disease,which is a clinical common,difficult to cure,and a multiple chronic inflammatory disease.It belongs to a kind of autoimmune disease.Its morbidity number accounts for about 2%?3%of the total population of the world.Although the pathogenesis of psoriasis is not very clear,a large number of studies have shown that the inflammatory immune response dysfunction in the skin tissue is one of the main causes of the lesion formationPrevious studies reported that under the action of chemotactic factor CCL20,CCR6+Th17cells,which are the main effector cells in the formation of "vicious circle" of psoriasis immune mechanism,move to the skin lesion,where they produces inflammatory cytokines such as IL-17A,IL-22,IL-17F and TNF-?.However,in recent years,more and more clinical and basic studies have proved that the skin dermal ??T cells are also an important source of cytokines,such as IL-17A and IL-22,and they are also extensively involved in the occurrence and development of psoriasis.The immunologic role of the skin ??T cells has gradually become a new focus in the basic and clinical researches of the pathogenesis of psoriasisTo investigate the mechanism of CD30L/CD30 signal transduction in the formation of skin T17 cell-mediated psoriasis,this study used CD30L knockout mice to establish the Imiquimod(IMQ)induced vulgaris psoriasis animal models,and futher,the regulation effect of CD30L/CD30 signal transduction on T17 cell-mediated skin inflammatory immune response was discussed in detail.Meanwhile,the therapeutic effect of CD30L/CD30 signal on psoriasis mice was evaluatedMethods:1.using C57BL/6 mice to establish the animal model of psoriasis:take mice skin lesion tissue,extract the total protein,and use WB technology to detect the expression changes of CD30L protein in skin lesion tissue.Cd30l gene knockout mice(CD30LKO)were used to establish IMQ-induced psoriasis animal models,and the effects of Cd30l gene deletion on the occurrence and development of psoriasis skin inflammation in mice were dynamically compared and observed2.WT and CD30LKO mice were used to establish animal models of psoriasis:Detect Infiltration of neutrophils and macrophages in mouse dermal tissue and dLN;CD4+or CD8+??TCR+T cell;the levels of CCR6 and Ki-673.In vitro experiments were used to dynamically compare and analyze the relationship between CD30L and the expression of CCR6 in the subgroup of ??T cells4.The function of the ??T cell in dermal tissue and dLN,especially the function of V?4+?? T cell,the expression of IL-17A?IL-22 or IL-17F.5.Stimulating monoclonal antibodies were used to target CD30L/CD30 signals to evaluate the effect of CD30L/CD30 signal regulated by monoclonal antibody on mouse psoriasis.Results:1.Deletion of Cd30l gene resulted in significantly increased susceptibility to IMQ induced psoriasis in mice.The results showed that the deletion of Cd30l gene resulted in obvious aggravation of the mice skin inflammation induced by IMQ,and the mice were significantly susceptible to psoriasis,which suggested that CD30L/CD30 signal transduction play an important and protective role in the pathogenesis of psoriasis2.Cd30l gene deletion caused inflammatory cell infiltration and enhancement in mice skin lesions.Our results showed that Cd30l gene deletion can promote the CDllb+Gr-l+neutrophils which are in the dLN of mice expression of CXCR2 to infiltrate the tissue,thereby promoting the formation:vicious circle" of psoriasis3.Deletion of Cd30l gene led to the enhancement of the V?4+??T cells homing in the skin,and thus promoted the infiltration of CXCR2+neutrophils into the dermal tissue4.Cd30l gene deletion led to increase in the expression of CCR6 and Ki-67 by CD27-V?4+??T cells.The number of homing V?4+??T cells in the skin lesion of CD30L KO mice were increased,thus promoting the formation of the "vicious cycle" of psoriasis5.CD30L/CD30 signal transduction can inhibit the expression of the CCR6 of V?4+??T cells:The deletion of CD30L/CD30 signal transduction caused the activation V?4+??T cells of the dLN in psoriasis mice continuous expression of high levels of V?4+??T CCR6,Furthermore,a large number of lesions were migrated to dermal tissue to mediate the inflammatory response of the skin6.Cd30l gene deletion resulted in increased activation of CCR6+V?4+??T17 cells The infiltration of neutrophils were induced by the production of a large number of IL-17A,IL-22 or IL-17F etc.and other related cytokines,and furthermore,the process of psoriasis mice was exacerbated.7.Evaluation of the efficacy of CD30L/CD30 signal targeting antibody against psoriasis WT mice:the efficacy of psoriasis WT mice results fully proved that monoclonal antibody stimulation CD30L/CD30 signal transduction can inhibit V?4+??T cells and skin homing ability related to the cytokines secreted by ??T17 cells,partly inhibiting the formation of the mice psoriasis "vicious circle",which has significant therapeutic effect on psoriasis mice.Therefore,CD30L/CD30 signal may be a potential candidate target for the treatment of psoriasis.Conclusion:1The interaction between CD30L and CD30 moleculeson the surface skin homing V?4+??T17 cells can inhibit V?4+??T17 cells express CCR6,down-regulated the production of inflammatory cytokines such as IL-17A,IL-22 and IL-17F,indirect inhibition of neutrophil migration to skin tissue,and slow down the formation of the"vicious circle" in the pathogenesis of psoriasis.2.CD30L/CD30 signaling effected in psoriasis mice by using CD30L/CD30 signal targeting monoclonal antibody.3.CD30L/CD30 signal transduction played an important protective role in the formation of psoriatic lesions mediated by ??T17 cells.