Analysis Of Lymphocyte Subsets In Pediatric Systemic Lupus Erythematosus And Was Protein Defects Leads To The Intrinsic Defects Of The Follicular Helper T-cell

PART I ANALYSIS OF LYMPHOCYTE SUBSETS IN PEDIATRICSYSTEMIC LUPUS ERYTHEMATOSUSObjective: To analyze in the number of lymphocyte subsets and distribution in pediatric systemic lupus erythematosus(p SLE)and further explore the role of the lymphocyte subsets in the plausible pathogenesis of p SLE in order to provide evidence for early diagnosis and treatment of p SLEMethods: Peripheral blood samples of 29(female: male = 6.5: 1)p SLE patients from March 2015 to December 2016 were enrolled,including 15 cases of disease onset group,14 cases of disease stable group and 15 age-mathced healthy children as controls.Flow cytometry was used to detect the frequency and number changes of γδT cell,Tfh cell and Treg subpopulations in p SLE in different disease periods.T cell,B cell and NK cells and the proportion of T and B cell subsets in peripheral blood ofchildren were analyzed comprehensively on the basis of fine immunophenotyping method of whole blood lymphocyte with flow cytometry.Results: The proportion and the absolute number of T cells in the stable group were higher than those in the control group and the disease onset group.Meanwhile,the proportion and absolute value of T cells in the disease onset group were lower than those in the control group.The proportion and absolute value of CD8 + T cells in the stable group were higher than those in the disease onset group.The proportion of B cells in the disease onset group was higher than that in the control group and the stable group.There was no significant difference between the three groups.The proportion of CD4 + cells in the stable group was significantly higher than those in the disease onset group and the control group,and the absolute value of CD4 Naive was lower than that of the control group.There was no significant difference in the proportion and absolute number of the other Th cells.The proportion and absolute value of CD8 TEMRA in the stable group were higher than those in the health control group.The absolute number of CD8 Naive in the disease onset group was lower than that in the normal group,and the proportion of the remaining Tc subpopulation and the absolute number were not significantly different between the three groups.The percentage and absolute number of Na?ve cells in stable group were lower than those in disease onset group and thecontrol group.There was no significant difference in the proportion and absolute number of the other B cell subsets between the three groups.The proportion of γδT in the disease onset and stable groups was significantly lower than that in the control group.The proportion of Tfh cells,ICOS and PD-1 expression in SLE were significantly lower than those in the control group,and the difference was statistically significant.The proportion of non-Treg(non-Treg)cells in the disease onset and stable groups was significantly higher than those in the control group.The proportion of activated Treg(a Treg)cells in the disease onset group was higher than that in the stable group.There was no significant difference in the resting Treg between the three groups.Conclusion: The proportion of Tc,Th and B cell subsets in peripheral blood of p SLE patients was imbalance,suggesting that the proportion and the absolute number of immune cells may be involved in the pathogenesis of p SLE.Additionally,γδT cells in p SLE decreased significantly,suggesting that γδT may play an important role in its pathogenesis of p SLE.In addition,non-inhibitory Treg(non-Treg)cell ratio was significantly higher than the controls,indicating that the decline of immunosuppressive function in children,which result in excessive immune response,and finally leading to the occurrence of p SEL.Moreover the decrease in the number of c Tfh cells in the stable group suggests that Tfh cells may be used as a clinical indicator to reflect the efficacy of treatment,to assess theoutcome and prognosis of children.PARTⅡWAS PROTEIN DEFECTS LEADS TO THE INTRINSICDEFECTS OF THE FOLLICULAR HELPER T-CELLObjective: In our previous study,frequency,absolute number and function of Tfh was demonstrated impaired in the absence of WASp expression either in WAS patients or WASKO mouse models.The aim of this study was to determine whether the above abnormalities of Tfh cells in WASKO mice was an intrinsic defect caused by WASp deficiency by bone marrow mimicking rat animal model and to demonstrate whether Tfh cell defects are inherent in WASp deficiency to enhance the understanding of the pathogenesis of the disease,and ultimately provide a new possible direction for the clinical treatment of children with WAS.Methods: Bone marrow cells of WASKO mice / WT mice(expressing CD45.2)were mixed with CD45.1 mouse(WT mice)bone marrow cells 1:1 and transplanted into lethal dose irradiated CD45.1 mice(WT small Mice)to establish a bone marrow mimetic rat model.After 8 weeks of stem cell remodeling,the mice were immunized with NP-KLH.After 2 weeks,the percentage of CD45.2 positive Tfh cells,ICOS + Tfh cell ratio,PD-1Tfh cell ratio,and Tfh cell proliferation and Apoptosis were determined.Results: The proportion of CD45.2 + Tfh cells in WASKO group was significantly decreased compared with that in the control group.Meanwhile,the proportion of ICOS + Tfh cells,the proportion of PD-1 + Tfh cells and the proliferation level of Tfh cells were decreased,whereas the apoptosis of Tfh cells was increased.Conclusion: This study have found that when WAS protein defected,the proliferation level of Tfh cells were significantly reduced and the apoptosis was increased in WASKO mice,indicating that WASp is closely related to the production of Tfh.