Expression And Significance Of Th17 Cells From Peripheral Blood Of Patients With Inflammational Arthritis

Objective:(1)To study expression and significance of Th1,Th2,Th17 cells from peripheral blood of patients with inflammational arthritis(rheumatoid arthritis and ankylosing spondylitis).(2) To study expression of IL-17,RORγtmRNA in CD4~+T cells from patients with inflammational arthritis(RA and AS).Methods:(1) The specimens of venous blood PBMC were collected from 48 patients with RA and 30 healthy persons. Intracelluar flow cytometirc detection of IL-17/IFN-γand IL-17/IL-6 was established using Anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4~+T cells from PBMC. The expression level of IL-17,RORγt mRNA in CD4~+T cells was determined from 18 RA patients and 15 healthy controls by real-time fluorescence quantitative RT-PCR using Anti-CD3/Anti-CD28/IL-23 as stimulators or not. There are three groups in the study named①healthy controls group;②RA stable group;③RA active group。(2) The specimens of venous blood PBMC were collected from 40 patients with AS and 15 healthy persons. Intracellular flow cytometry detection of IL-17/IFN-γand IL-17/IL-6 was established using Anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4~+T cells from PBMC. The expression level of IL-17. RORyt mRNA in CD4~+T cells was determined from 28 AS patients and 15 healthy controls by real-time fluorescence quantitative RT-PCR using Anti-CD3/Anti-CD28/IL-23 as stimulators or not. There are three groups in the study named (1)healthy control group; (2)AS stable group; (3)AS active group。Results:The isolation of untouched human CD4+T cells from PBMC was effective and its purity was over 90%. The percentage of intracelluar IL-17 in CD4+T cells from RA and AS patients was increased significantly. Such percentage in active group was higher than that of stable group(p<0.01) and both of them were higher than that of healthy controls(p<0.01). Under Anti-CD3/Anti-CD28/IL-23 stimulation, the percentage of intracelluar IL-17 in RA patients was increased significantly(p<0.01).But there is no significant statistical significance in AS patients before and after the stimulation (p>0.05). There is no correlation between IL-17 and IFN-γor IL-6. The expression level of IL-17,RORγt mRNA in CD4+T cells was significantly higher in patients with RA and AS than that of the controls. Under Anti-CD3/Anti-CD28/IL-23 stimulation, the percentage of IL-17,RORγt mRNA was increased significantly (p<0.01). Such percentage of IL-17,RORytmRNA in 12h group was higher than that of 24h group, while both of them were higher than that of Oh controls(p<0.05).Conclusion:There is an abnormal expression of IL-17 in human CD4+T cells from RA and AS patients, which suggested that Th17 cells could contribute to the pathogenesis of RA and AS.The abnormal high expression of IL-17 might play a role in the development and progression of RA and AS.