Vγ4~+γδ~+T Cells Affect The IMQ-induced Paosiriasis-like Inflammation

BackgroundPsoriasis is a common disease and the psoriasis vulgaris is the most common,which clinical performance presents scales erythema and shows reddish shiny transluce membrane when scrapped away,then the little bleeding spot will occur if the membrane is broken.There are many pathogenic fators to lead the occurrence of psoriasis such as genicinfection,endocrine disordors,abnormal immune system.The lesions are often occurred in face,trunk and extremities,which will seriously affect the patient’s physical and mental health.T cells play an important role in human cell-mediated immunity,including the idtentifiacation of antigens,differentiation of effector cells and memory cells.the effecor cells can secrete lymphatic fator and combine with target cells to give role in immune,and the memory cells can differentiate into the effecor cells.The functions of T cells can protect the human body from infectious,physical,and chemical damage.So the abonormal immune system of T cells may lead to autoimmune disease.Up to date,there are so much research focusing on the disease of immune system,but the role of Vγ4T cells in epidermis have not been reported.In this study,we explored the function of Vγ4~+γδ~+T cells in epidermis and the change of dermal Vγ4~+γδ~+T cells and cytokines-produing Vγ4T cells in re-challenged mice by the Vγ4T cells-depleted mice.As previous reports have confirmed that the Vγ4T cells strengthen the psoriasis-like inflammation,we predict that Vγ4T cells-depleted mice relieved the IMQ-induced psoriasis-like inflammation both in primary and re-challenged mice.ObjectiveWe confirm that the Vγ4T cells play a role in the model of IMQ-induced psoriasis-like inflammation,by building the different conditons of mice,which may provide a new idea for the clinic to find out the exact mechanism of psoriasis.1、 grouping and building the model of Vγ4~+T cells-depleted mice.Six to eight weeks female C57BL/6 mice were randomly divided into two groups,Vγ4~+T cells-depleted group was obtained by intraperitoneal injection of anti-Vγ4 antibodies three days before the IMQ-induced psoriasis-like inflammation,and the mice of wild-type group was injected the same volume PBS-buffer.The dermal Vγ4~+γδ~+T cells could not be detected by FACS,indicating that the model of Vγ4~+ T cells-depletedmice was successful.2、 Building the model of the IMQ-induced psoriasis-like inflammation.Six to eight weeks female C57BL/6 mice received 5% IMQ cream for 6 successive days to induce the psoriasis-like inflammation.The severitiy of the erthyma,scales and thickness were evaluated by the PASI standard and the ImageJ was used for measuring the thickness of the epidermal tissue.3、 Vγ4~+γδ~+T cells in the epidermis tissues have an effect on psoriasis-like inflammation and the mechanism research.3.1 the expression of IL-17 was dectected by western blotin the epidermial tissue of psoriasis-like inflammation,the FACS was used as the dectection of IL-17-producing γδT cells and Vγ4~+γδT cells and IL-17-producing Vγ4~+Tcells.3.2 the severity of psoriasis-like inflammation in both wild-type andVγ4Tcells –depleted mice was evaluated by the PASI scoring and pathological change of HE staining.Using the FACS to detect the Vγ4~+γδT cells and IL-17-producing cells of the epidermis in the Vγ4T cells–depleted mice.3.3 The expression of IL-23,IL-1βwere determined by immunohistochemical staining inthe epidermal tissues of psoriasis-like inflammation and the health and psoriasis patient.Compared the IMQ-induced psoriasis-like inflammation between IL-23,IL-1β neutralizing mice and wild-type mice.4、 the dermal Vγ4~+γδ~+T cells have an effect on re-challenged psoriasis-like inflammation and the mechanism research.4.1 the severity of psoriasis-like inflammation in both primary and re-challenged mice was evaluated by the PASI socre standard and pathological change of HE staining.Using the FACS to detect the dermal Vγ4~+γδT cells and IL-17 producing Vγ4~+γδT cells in the mice of two groups.Method4.2 Compared the IMQ-induced psoriasis-like inflammation between re-challenged wildtype mice and wild-type mice,Vγ4-depleted re-challenged mice.Result1、 The models of the Vγ4-depleted mice were obtained successfully.2、 the wild-type group had a set of symptoms in the hair removal area like erythema,scales after daubing IMQ cream on the dorsal skin once a day for 6 successive days.and the PASI score of the wild-type mice was higher than the Vγ4T cells eliminated group.The acanthosis,parakertosis and the dermal infiltration of inflammatory cells were more obviously in the wild-type mice than the Vγ4-depleted mice by observation of hematoxylin and eosin staining sections.3、 Flow cytometry instrument detection showed both the epiderimis cells didn’t express Vγ4 T cells in the normal mice of wild-type group and Vγ4-depleted group,however,the Vγ4 T cells were increased to 20% of epidermal cells when we applied IMQ to induce psoriasis-like inflammation.In addition,the 40% of the Vγ4 T cells prodeuced IL – 17 A.however,in the Vγ4-depleted group,the Vγ4 T cellshardly occurin the epidermis and the IL–17A-producing cells were significantly reduced.4、 Western blot results found that the cytokinesof IL-23、IL-1β were expressed in the protein of epidermal tissue;Furthermore,theIMQ-induced psoriasis-like inflammation were relieved and the epidermal expression of IL-17 was less in the IL-23/IL-1β neutralizing mice.5、 the inflammation was more serious in re-challenged mice than primary mice and the dermal Vγ4~+γδ~+T cells responded quickly to the same IMQ stimulus.Therefore,that the Vγ4T cells secreted IL–17A、γ-IFN、IL-22 were increased in the re-challenged mice.6、 Depleting Vγ4T cells of the mice that once was induced psoriasis-like inflammation by IMQ before one month,in which relieved the IMQ-induced re-challenged psoriasis-like inflammation.And the expression of IL–17A、γ-IFN were decreased in the dermal cells,however,IL-22 was increased to compare with the re-challenged wild-type mice.Conclusion1、 The IMQ-induced psoriasis-like inflammation was compared in wild-type and Vγ4T cells-depleted mice,the Vγ4T cells occured in the epidermis and secreted IL-17 and the psoriasis-like inflammation was relieved in the Vγ4T cells-depleted mice.2、 The cytokines of IL-23,IL-1β in the epidermis play a role in the IMQ-induced psoriasis-like inflammation.3、 The dermal Vγ4T cells of the mice that was once induced inflammation rapidly responded to the same stimulus and proliferated and secreted cytokines to promote the development of inflammation.In this study,we confirmed that the Vγ4T cells of the skin play an important role in the pathogenesis of psoriasis,which may provide a new guiding idea for the clinic to dermine the definite mechanism of psoriasis.