Study On Langerhans Cells In The Uptake Of Allergen And The Induction Of Immune Inflammation Based On Skin Single Cell Preparation

Objective To investigate the differences in level of eosinophilic granulocyte(EO)and serum IgE between atopic dermatitis(AD)group and healthy control group,to explore the correlation with the severity of the disease,and to analyze whether it can be used as potential diagnostic biomarkers of AD.To improve theefficiency of current methods for the separation of LCs and obtain LCs withhigher quality,and to confirm the capacity of Langerin that expressed on the surface ofepidermal LCs to bind multiple polysaccharide antigens;moreover,to explore theroles of LCs in the process of capuring and presenting polysaccharide antigens and the functions of LCs to induce of the differentiation of T cells.Methods From Jan 2016 to Aug 2016,blood from 154 patients with AD and 317 healthy people were collected,and were divided into AD group and control group.SYSMEX SE-9500 automatic hematology analyzer and ImmunoCAP system were used to detect the level of EO and IgE in two groups.SPSS 20.0 software was used to analyze the differences of EO and IgE levels between the two groups,as well as the correlation with SCORAD score.ROC curve of EO and IgE in two groups was performed,and the areas under the curve were calculated.The statistical difference was defined as P<0.05.Comparing the capability of LCs to reconinize several polysaccharide antigens before and after maturation under the local environment(TSLP);comparing LCs that separted from human skin using skin patching technology and single cells suspension technology by evaluating the cell vialibity and the expression of Langerin on the surface of LCs;investigating the effect of the activated LCs on the polarization of T cells.ResultsThe levels of EO in AD patients and healthy people were(0.40±0.03)×109,(0.13±0.01)×109.The levels of serum IgE in AD patients and healthy people were(684.4±95.22),(44.24±1.37)IU/ml.In addition,the levels of EO and IgE in AD patients were both associated with the SCORAD score(P<0.001,P<0.01).The areas under ROC curve of EO and IgE in groups were 0.76,0.79(P<0.001),respectively.Compared with Blank,the expression of CD80,CD83,CD86,HLA-DR were significalty increase with the stimulation of TSLP + derp,adding derp,OVA or alpha-gal(P<0.05);moreover,compared with cells that seprated using single cell suspension technology,LCs seprated using skin patching techbology has shown significant cell viability[(94.6±14.2,respectively.)%v.s.(76.3±13.1)%(P<0.01)],and also increased the positive rate of CD207[(16.8±4.7)%v.s.(12.1±3.5)%(P<0.01)];furthermore,the concentrations of IL-4 in supernatants of derp group and Blank group were(368.4±73.1)and(195.3±pg/ml 53.7)pg/ml,,and the concentrations of IFN-gamma were(278.6±79.2)pg/ml and(631±103.5)pg/ml,suggesting that compared with the Blank group,the concentration of IL-4 in supernatant of derp group increased significantly(P<0.01),and concentration of IFN-gamma decreased markedly(P<0.005).Conclusion The levels of EO and IgE in patients with AD were significantly higher than that of healthy people,and they may have a certain instruction to the disease,which are expected to be potential diagnostic biomarkers.Skin patching technique is superior to single cell suspension technique,and Langerin is involved in the process of recogniation,caupure and presentation of OVA(derp)and-gal;moreover,LCs plays a key role in the differentiation of T cells.