The Role Of T Cell Receptor Regulates CD4 T Cells Differentiation In Airway Inflammation Of Asthma

Objective: To interve tumor necrosis factor receptor 2 in eosinophilic asthma and aryl hydrocarbon receptor in non-eosinophilic asthma and observe their effects on this two kind of airway inflammation.Methods:(1)eosinophilic asthma model was induced by OVA,TNFR2 mRNA and protein of lymphocytes derived from spleen and lung drainage lympho node were detected by real-time PCR and western blot respectively.(2)Inhibited TNF-TNFR2 signal by TNFR2 blocking peptide administration intranasally.Lung histologic HE staining and PAS staining,total cell count and differential cell count of BALF,cytokine assay of serum and BALF were used for estimated the effect of TNFR2 inhibition on airway inflammation.Differentiation of CD4 T cell in vivo and in vitro was analyzed by flow cytometry.The transcript factors of CD4 T cell were detected by western blot.(3)Active TNFR2 signal by TNF and TNFR1 antibody administration.Lung histologic HE staining and PAS staining,total cell count and differential cell count of BALF,cytokine assay of serum and BALF were used for estimated the effect of TNFR2 activation on airway inflammation.Differentiation of CD4 T cell in vivo and in vitro was analyzed by flow cytometry.The transcript factors of CD4 T cell were detected by western blot.(4)non-eosinophilic asthma mice model was induced by OVA and LPS.TCDD gavage was used for Ah R activation.(5)Total cell count and differential cell count of BALF,lung histologic HE staining and PAS staining,immunohistochemical staining of MPO and Gr-1 were used for airway inflammation estimation.Differentiation of Treg and Th17 were analyzed by flow cytometry,Fox P3 and RORγ protein expression were detected by western blot.(6)dendrite cells from lung were isolated by magnetic beads,CD80 and CD86 expression on DC were analyzed by flow cytometry.Results:(1)m RNA and protein expression of TNFR2 were decreased in spleen and lung lymphocytes from asthma mice.(2)TNFR2 inhibition aggravated airway inflammation as well as inflammation cytokine expression of asthma mice.TNFR2 inhibition promoted Th2 and Th17 differentiation in vivo and in vitro as well as the GATA3 and RORγ expression but inhibited Treg and Th1 differentiation and Fox P3 and T-bet expression.(3)TNFR2 signal activation reduce the airway inflammation of asthma mice.TNFR2 signal activation inhibited Th2 and Th17 differentiation in vio and in vitro as well as GATA3 and RORγ expression but promoted Treg and Th1 differentiation as well as Fox P3 and Tbet expression.(4)Ah R activation reduced the airway hyper reactivity and alleviated airway inflammatory cells especially neutrophils infiltration.Ah R activation reduced the expression of IL-4 and IL-17 in serum and BALF of asthma mice.Ah R activaton promoted Treg differentiation and Fox P3 expression but inhibited Th17 expression and RORγ expression.(5)Ah R activation has no influence on CD80 and CD86 on DC from asthma mice.Conclusion: TNFR2 signal activation reduce the airway inflammation of asthma mice that may associated with its regulation function on differentiation of CD4 T cells.Ah R activation alleviated airway inflammation of non-eosinophilic asthma via regulating Treg and Th17 differentiation,but that may not associated with maturity of DC.