Research On The Relation Between ILC2,B10 Activity And Autoantibodies In Hashimoto's Thyroiditis Patients

Objective: Hashimoto thyroiditis(HT)belongs to organ specific autoimmune diseases.Antithyroglobulin antibody(ATG)and anti-thyroid peroxidase antibody(ATPO)are significantly increased in HT patients and contribute to the pathogenesis of HT.However,the detailed mechanisms remain unclear.In the present study,group II Innate lymphoid cell(ILC2)specific transcription factor,retinoid-related orphan receptor ?(ROR?),and cytokines,IL-5 and IL-13 were detected.Correlation analyses between ROR?,IL-5,IL-13 and ATG,ATPO were performed to explore the relation between ILC2 activity and autoantibodies in HT patients.In the meantime,peripheral blood B10 cell specific cytokine IL-10 was detected.Correlation analyses between IL-10 and ATG,ATPO were performed to explore the relation between IL-10 and autoantibodies in HT patients.Methods: 1.30 patients first diagnosed with HT in Jiangsu university affiliated people's hospital were selected.Among them,26 cases were female and 4 cases were male with an average age of 41±14.All patients were diagnosed accord with the China guide of diagnosis and treatment of thyroid disease.30 healthy physical examinees without thyroid disease,autoimmune disease and related family history were selected as control group.Among them,25 cases were female and 5 cases were male with an average age of 44±10.2.Immunochemistry luminescence method was performed to analysis plasma ATG,ATPO,FT3,FT4 and TSH with LDX800 machine(Beckman Coulter).3.Ficoll-Hypaque density gradient centrifugation method was to isolate peripheral blood mononuclear cells in HT patients.Total RNA was extracted and OD260/280 was detected by spectrophotometer to analysis the concentration and purity.4.Real-time PCR was performed to detected m RNA expressions of ILC2 specific transcription factor(ROR?),cytokines(IL-5 and IL-13)and B10 cell specific cytokine(IL-10).5.Enzyme-linked immuno sorbent assay(ELISA)was performed to detected plasma ILC2 specific cytokines(IL-5 and IL-13)and B10 cell specific cytokine(IL-10).6.Graphpad5.0 software was used for statistical analysis.Non-paired student-t analysis was conducted to compare differences between two groups.Pearson analysis was conducted for correlation analyses between two groups.Results: 1.Plasma ATG,ATPO and TSH were significantly increased,while FT3 and FT4 were significantly decreased in HT patients,which presented the obvious pathological characteristics of HT.2.Results of RT-PCR showed that m RNA expressions of ILC2 specific transcription factor(ROR?)and cytokines(IL-5 and IL-13)were significantly increased in HT patients,which indicate that ILC2 activity was promoted in HT patients.B10 cell specific cytokine(IL-10)was significantly decreased in HT patient.3.Results of ELISA show that plasma ILC2 specific cytokines(IL-5 and IL-13)was significantly increased,while B10 cell specific cytokine(IL-10)was significantly decreased in HT patient.4.Results of correlation analyses showed a significant positive correlation between plasma ROR? and ATG,ATPO.A significant positive correlation was also seen between plasma IL-5 and ATPO.However,a slight but not significant positive correlation was seen between IL-5 and ATG as well as IL-13 and ATG,ATPO.Results of correlation analyses showed a significant negative correlation between IL-10 and ATPO and a slight but not significant negative correlation between IL-10 and ATG.Conclusions: Increased thyroid autoantibodies in HT patients have a close relationship with enhanced ILC2 activity and decreased IL-10 level,which may be the important immunological mechanisms of HT pathogenesis.