Study On Treg From Peripheral Blood Of CD Patients And Function Of LncRNA DQ786243

Objective: Study focuses on Treg in peripheral blood, research on Treg function,phenotypic changes in Crohn’s disease patients. And also, the impact of lncRNADQ786243on the expression of foxp3, the control gene of Treg. Analyze therelationship between the quantity of Treg and characteristics of CD patients.Methods: Use magnetic beads to sort CD4+CD25+Treg and CD4+CD25-Teff. WhenTreg from patients and healthy controls co-cultured with Teff, we use CFSE dilutionmethod to tell the difference of Treg suppression function. We use fluorescent antibodylabeling CD4+T cells to detect differences in the number of CD4+Foxp3+marked cellsand CD4+CD45RA-Foxp3+marked cells in patients and healthy controls. Detect theexpression of lncRNA DQ786243, CREB and Foxp3between patients and healthycontrols by qRT-PCR, and perform correlation analysis with clinical data. ObserveCREB, Foxp3expression changes, and changes in levels of CREB phosphorylation inJurkat cells transfected with lncRNA DQ786243. In a larger amount of samples, detectperipheral Foxp3TSDR demethylation level in CD patients and healthy controls, tounderstand the differences in the number of Treg in CD patients and healthy controlgroup, as well as differences in different therapy and different clinical features of theCD.Results：When co-cultured Treg and Teff, CD patients Teff average growth ratecompared with the control group within24hours, there was no statistically significantdifference. The proportion of CD4+CD45RA-Foxp3hiaTreg decreased in patients withactive CD (P <0.01). Quantitative RT-PCR showed that the expression DQ786243inactive CD patients was significantly higher (P=0.004). CRP and DQ786243there is agood correlation (r=0.489, P=0.034). There is a certain linear correlation betweenexpression of Foxp3and DQ786243(r=0.435, P=0.021). No significant correlationbetween expression of CREB and Foxp3. Forty-eight hours after DQ786243transfection, Foxp3mRNA expression in Jurkat cells was significantly high (P=0.046),p-CREB/t-CREB in24hours and48hours post-transfection increased (P=0.0043). Comparison found no significant differences in TSDR demethylation proportion ofpatients and healthy controls. Proportion of TSDR demethylation elevated in patientswith perianal disease (T-Test, P=0.0036). After DQ786243transfection TSDRdemethylation levels appeared elevated and statistically significant (ANOVA, P=0.0010).Conclusion: There was no significant difference in suppressive function of Treg innormal controls and patients with CD. Using CD4+CD45RA-Foxp3hias active Tregmarker, it found that amount of Treg is lower in patients with CD than in controls.DQ786243is related with disease activity and can affect the expression of CREB andFoxp3. CREB itself does not mediate the regulating of Foxp3directly, it might workwith the phosphorylation of CREB. There is no sufficient evidence to show that thereare differences of the amount of Treg in peripheral blood between patients and CD. Theamount of Treg in CD patients with perianal lesions appeared elevated. In Jurkat cellstudy, DQ786243can affect Foxp3TSDR demethylation level, which may be anothermechanism of Foxp3regulation in vivo.