Study About Different Doses Of Standardized Dust Mite Vaccine Effcet On Foxp3~+CD4~+ CD25~+ Regulatory T Cells From Astthmatic Model Mice

Objective: To establish dermatophagoides farinae (der f) allergic asthma model in mice and give the asthmatic mice der f vaccine for specific immunotherapy, from both mouse pathology and cytokines to reflect der f vaccine treatment, and to compare different doses of der f vaccine in the treatment of effect on Foxp3~+CD4~+CD25~+ regulatory T cells, and to find the best dose of der f vaccine for treatment.Methods:32 female C57BL / 6 mice were randomly divided into 4 groups after setting up the allergic asthma model with the extract of der f. Normal saline group given der f allergen extract stimulated ~+normal saline treatment; low-dose group given der f allergen extract stimulated ~+ low-dose der f extract treatment; medium-dose group given der f allergen extract stimulated ~+ medium-dose der f extract treatment; high-dose group given der f allergen extract stimulated ~+ high-dose der f extract treatment. After treatment, levels of CD4~+CD25~+T cells and Foxp3~+CD4~+CD25~+Tregs in spleen cells of mice were measured by flow cytometry. Serum levels of interleukin-10 and Transforming growth factor-β1 were detected by ELISA.Results:1.The ratio of CD4 ~+ CD25 ~+ T cells and CD4 ~+ T cells in spleen cells of mice compared normal saline group, low-dose group respectively [(7.49±0.72)%, (8.87±0.33)%] with medium-dose group [(18.23±0.58)%] ,there was statistically significant different (P <0.05), while medium-dose group compared with high-dose group[(15.23±1.50)%] , there was no significantly statistical different(P> 0.05); The ratio of Foxp3~+CD4~+CD25~+Tregs and CD4~+CD25~+T cells compared normal saline group, low-dose group and group respectively [(7.78±0.36)%, (8.71±0.46)%] with medium-dose group [(13.51±1.39)%] there was significant different(P <0.05,) while medium-dose group compared with high-dose group [(12.12±1.44)%] , there was no significantly different (P> 0.05).2.Normal saline group, low-dose group about the serum levels of IL-10 respectively [ (48.14±0.38) ng / l, (50.01±0.93) ng / l], the serum levels of TGF-β1 respectively [ (72.41±1.65) ng / l, (74.67±1.66) ng / l] was lower than medium-dose group [(65.13±0.61) ng / l , (102.43±1.77) ng / l], there was significantly different (P <0.05), while medium-dose group increased comparing with high-dose group[(63.91±0.27) ng / l, (100.11±1.26) ng / l] , but there was no statistically significantly different (P> 0.05). Conclusion:1. This study successfully established der f asthmatic model and completed of different doses of der f vaccine in SIT,and found SIT could increase the expression of Foxp3, and promote the secretion of IL-10 and TGF-β1.2. Dose of Der f vaccine could effect on the number and function of Foxp3~+CD4~+CD25~+ regulatory T cells research ,dose of Der f vaccine and the number and function of Foxp3~+CD4~+CD25~+ regulatory T cells was not proportional, and found that medium-dose dust mite vaccine was the most significant on immunotherapy, so the best dose of immunotherapy was medium-dose.