Altered Expression Of TLR9 In Blood B Cells And Treg Cells Of Allergic Rhinitis,Asthma

Objective To investigate the expression of Toll-like receptor 9 in peripheral blood B cells and regulatory T cells in patients with allergic rhinitis,allergic asthma,allergic rhinitis combined with asthma airway allergy.To detect the effects of different allergens on TLR9 expression in peripheral blood B cells and Treg cells;A sensitized mouse model was constructed to explore the effects of allergens and FcεRI receptors on TLR9 expression on B cells and Treg cells.To provide data for further exploration of the potential role of TLR9 in AR,AA,ARA.Methods The peripheral blood of a total of 100 volunteers(19 healthy people,40 AR patients,26 AA patients,and 15 ARA patients)was recruited;Construction of sensitized wild type(WT)mice and FcεRI receptor knockout(FcεRI-KO)mouse models;Tested by flow analyzer:1.Detect the expression and MFI of TLR9 in peripheral blood B cells of patients with HC,AR,AA and ARA.The expression and MFI of TLR9 in volunteer peripheral blood B cells and Treg cells before and after the excitation of different allergens(ASWE,HDME,PPE)were detected.2.To detect the expression of TLR9 and MFI in peripheral blood Treg cells of patients with HC,AR,AA and ARA.The expression ratio and MFI of TLR9 in Treg cells of volunteers before and after the excitation of different allergens(ASWE,HDME,PPE)were detected.3.To detect the expression of TLR9+ in blood CD19+ B cells of WT mice and FcεRI-KO mice sensitized by ASWE and HDME.Observe and compare the experimental results of WT mice and FcεRI-KO mice.4.To detect the expression of TLR9+ in blood CD4+CD25+Foxp3+ Treg cells of WT mice and FcεRI-KO mice sensitized by ASWE and HDME.Observe and compare the experimental results of WT mice and FcεRI-KO mice.Results1.TLR9+ expression results of CD19+ B cells in human peripheral blood:The expression percentage of TLR9+ B cells in peripheral blood of AR patients,AA patients and ARA patients was higher than that of HC TLR9+ B cells,and the corresponding mean fluorescence intensity(MFI)trend was also consistent.Before and after different allergen excitation,the expression percentage and MFI of CD19+ TLR9+ B cells in blood of patients with AR and AA were increased.2.TLR9+ expression results of human peripheral blood Treg cells:Compared with the HC group,the expression ratio of TLR9+Treg cells and MFI in peripheral blood of AR patients,AA patients and ARA patients showed a downward trend.After stimulation by different allergens,the proportion of expression of ASWE and HDME cells in peripheral blood of AR patients decreased compared with pre-stimulation after stimulation.The expression ratio of TLR9 in Treg cells after HDME stimulation was reduced in AA patients compared with before stimulation,and the TLR9 MFI decreased after PPE stimulation compared with that in Treg cells before stimulation.3.Blood CD19+ B cells of C57BL/6 mice TLR9+ expression:In the AR model,TLR9+ in the blood of CD19+ B cells of WT mice and FcεRI-KO mice was highly expressed and there was no significant difference.The MFI results showed that the TLR9+MFI of blood B cells sensitized by WT mice was higher than that of the NS control group,and the TLR9+ MFI of blood B cells sensitized by FcεRI-KO mice was also higher than that of the NS control group,but the overall TLR9+ MFI of WT mouse B cells was higher than that of FcεRI-KO mice.In the AA model,TLR9+ in the blood of CD19+B cells of WT mice and FcεRI-KO mice was highly expressed and there was no significant difference.WT mice and FcεRI-KO mice were higher in HDME-sensitized groups than B cells TLR9+ MFI in the NS control group,but WT mice were higher than FcεRI-KO mouse B cells TLR9+MFI.In lung tissues,FcεRI-KO mice had reduced TLR9+ expression and MFI in WT mouse B cells,and there were no significant differences before and after allergen stimulation in other groups except for the FcεRI-KO mouse HDME sensitization group.4.TLR9+ expression results of C57BL/6 mouse blood Treg cells:In the AR model,the MFI of TLR9+Treg cells after sensitization in WT mice was lower than that in the NS control group,and the MFI of TLR9+Treg cells after sensitization in FcεRI-KO mice was also reduced compared with that in the NS control group,but there was no significant difference in the AA model.In both the AR model and the AA model,the overall level of WT mice was higher than that of FcεRI-KO mice.There was almost no difference in TLR9+ high expression in AR mouse and AA mouse Treg cells.Conclusions1.Allergen stimulation human peripheral blood test conclusion:The experimental study found that the expression of TLR9+ and MFI of peripheral blood B cells of AR,AA and ARA patients increased,and the percentage of TLR9+ expression and MFI of Treg cells decreased,indicating that B cells and Treg cells could participate in the occurrence of AR,AA,ARA through TLR9.Different allergen sensitization test results may suggest that different allergens can affect the occurrence and development of allergic diseases to varying degrees.2.Experimental conclusion of C57BL/6 mice model:The experimental results show that B cells may play a role in AR and AA through TLR9,and Treg cells may play a role in AR through TLR9.The results of TLR9+ MFI in the blood of B cells and Treg cells in WT mice were higher overall than in FcεRI-KO mice that suggested that Ig E-mediated mechanisms may influence TLR9 expression in B cells and Treg cells.