| Background and ObjectiveAlopecia areata(AA)is a kind of hair loss without scar,which happens suddenly,and the local skin is normal.Alopecia areata may affect people's physical and mental health,which may be one of the important factors of anxiety and depression.At present,the etiology of alopecia areata is not very clear.For a long time in the past,it was thought that the genetic individual inducing factors led to the main CD8 driven Th1 cell autoimmune response attacking the hair follicle and causing hair follicle damage.The latest data show that regulatory T cells,??regulatory T cells,NKT10 cells and mast cells around hair follicles regulate lymphocyte subsets to maintain the physiological immunity privilege of hair follicles,CD8+T cells,NK cell subsets and so-called "unconventional" T cells(invariant NKT cells,??T cells,classic NK cells,type 1 innate lymphoid cells)produce a large number of IFN-y,which makes the privilege of hair follicle immunity collapse and damage the hair follicle.Regulatory T cells(Tregs)play an important role in the control of allergic,autoimmune,inflammatory,tumor immune and other immune responses,and can promote the growth of hair.CD39+Tregs is foxp3high effect memory Tregs,which has a stronger ability to inhibit the proliferation of conventional effect T cells than CD39-Tregs.Although in recent years,there have been studies on the proportion of Tregs in CD4+T-cell pool in peripheral blood of patients with alopecia areata,However,few people detect the absolute number of Tregs in the peripheral blood of patients with alopecia areata and some research results are contradictory.However,there are few reports on the detection of CD39+Tregs in the peripheral blood of patients with alopecia areata.Therefore,we detect the proportion of Tregs in CD4+T-cell pool and proportion of CD39+Tregs in Tregs pool in the peripheral blood of patients with alopecia areata,and also detect the absolute number of Tregs and CD3+Tregs,so as to verify and supplement previous studies.Interleukin-2 participates in the induction of immune response by stimulating the increment and differentiation of CD4+T cells and CD8+T cells,and participates in the immunosuppression by combining with CD25 which is highly expressed on Tregs.In the past,it was reported that the concentration of IL-2 in peripheral blood of patients with alopecia areata was increased,but some people used low-dose IL-2 to promote the recruitment of Treg to treat severe alopecia areata,but the reports about IL-2 in alopecia areata are still few,and few studies analyzed the correlation between IL-2 and CD39+Tregs in alopecia areata.Therefore,by detecting the levels of Tregs,CD39+Tregs and IL-2 in the peripheral blood of alopecia areata and healthy people,this experiment aims to explore the role of CD39+Tregs and IL-2 in the pathogenesis of alopecia areata,so as to enrich the research of Treg cells and IL-2 in the pathogenesis of alopecia areata.Materials and Methods1.Materials23 patients with active alopecia areata were enrolled in this study.They were hospitalized in the dermatology clinic of the First Affiliated Hospital of Zhengzhou University from November 2019 to February 2020.There are confirmed and complete outpatient case data,age 16-47 years old,average 30.71±4.46years old,15 males and 8 females;9 cases of single focal alopecia,8 cases of multifocal alopecia,4 cases of diffuse alopecia areata,1 case of ophiasis and case of alopecia Universalis.Randomly according to age,gender matching into the control group,the control group from the First Affiliated Hospital of Zhengzhou University Physical Examination Center.2.Methods5ml peripheral venous blood was collected on an empty stomach from the experimental group and the control group,and then the percentages and absolute number of CD4+CD25+Tregs,CD4+CD25+CD127+Tregs and CD4+CD25+CD127+CD39+Tregs and the concentration of IL-2 were determined by flow cytometry automatic analysis software of backman company in the United States.The obtained data were analyzed by kaluza software and FlowJo.3.Statistical analylsisSPSS21.0 was used to deal with the experimental data.Normally distributed measurement data are expressed as mean±standard deviation(x±S),independent semple t test is used to comparison between the two groups;the median(25%quantile,75%quantile)[M(P25,P75)]is used to show the non-normal distribution measurement data,Mann-Whitney U test is used for comparison between the two groups,the difference is statistically significant when P<0.05;Spearman correlation coefficient is used for correlation analysis,and graphpad prism 8.0.2 is used for drawing.Results1.The absolute number of CD4+CD25+Tregs in peripheral blood of patients with alopecia areata is 64.06±28.91/?l,and that of healthy control group is 33.76±15.20/?l.The absolute number of CD4+CD25+Tregs in peripheral blood of alopecia areata is significantly higher than that of healthy control group.The difference was statistically significant(P<0.05);the percentage of CD4+CD25+Tregs/CD4+T cells in peripheral blood of patients with alopecia areata was 8.46(5.3,8.46)%,the healthy control group was 5.57(4.9-8.29)%,The ratio of CD4+CD25+Tregs/CD4+T cells in peripheral blood of patients with alopecia areata was higher than that of healthy controls,the difference was not statistically significant(P>0.05).2.The absolute number of CD4+CD25+CD127-Tregs in peripheral blood of patients with alopecia areata is 55.89(43.73,94.07)/?l,the healthy control group is 33.80(19.68-47.84)/?l,and the absolute number of CD4+CD25+CD127-Tregs in peripheral blood of patients with alopecia areata was significantly higher than that of healthy controls.The difference was statistically significant in the healthy control group(P<0.05);the ratio of CD4+CD25+D127-Tregs/CD4+T cells in peripheral blood of patients with alopecia areata was 6.79(5.09-9.00)%,and the healthy control group was 6.12(4.13,6.78)%,the ratio of CD4+CD25+CD127-Tregs/CD4+T cells in peripheral blood of alopecia areata was higher than that of the healthy control group,the difference was not statistically significant(P>0.05).3.The absolute number of CD4+CD25+Tregs and CD4+CD25+CD127+Tregs in peripheral blood of patients with alopecia areata are strongly correlated(r=0.924,P<0.05),and also in healthy control groups(r=0.933,P<0.05);There is a strong correlation between the ratio of CD4+CD25+CD127+Tregs/CD4+T cells and the ratio of CD4+CD25+CD127+Tregs/CD4+T cells in peripheral blood of patients with alopecia areata(r=0.855,P<0.05),also in healthy control groups(r=7.06,P<0.05).4.The absolute number of CD4+CD25+CD127-CD39+Tregs in peripheral blood of patients with alopecia areata was 20.03(15.72,32.23)/?l,and the healthy control group was 8.19(5.78,12.78)/?l,the difference was statistically significant(P<0.05);CD4+CD25+CD127-CD39+Tregs in peripheral blood accounts for 44.07(19.58,62.90)%of CD4+CD25+CD127-Tregs in peripheral blood,and 23.36(14.96,47.73)%in healthy control group,The ratio of CD4+CD25+CD127-CD39+Tregs in peripheral blood of patients with alopecia areata was higher than that of healthy controls,but there was no statistical significance(P>0.05).5.The plasma IL-2 concentration in patients with alopecia areata was 0.45(0.38,0.65)pg/ml,and the healthy control group was 0.47(0.38,0.65)pg/ml.The plasma IL-2 concentration in patients with alopecia areata was lower than that in the healthy control group.The difference was not statistically significant(P>0.05).The plasma IL-2 concentration in patients with alopecia areata was negatively correlated with the absolute number of CD39+Tregs(r=-0.72,P<0.05).Conclusion1.The absolute number of Tregs and CD39+Tregs in peripheral blood of patients with alopecia areata is significantly increased,suggesting that Tregs and CD39+Tregs are involved in the regulation of immune inflammatory process of alopecia areata.2.CD39+Tregs in peripheral blood of patients with alopecia areata is negatively correlated with the concentration of IL-2 in plasma,indicating that the two regulate each other in the pathogenesis of alopecia areata and jointly participate in the immune response of alopecia areata. |
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