Study On The Expression Of TLR9 In Neutrophils And Monocytes In The Blood Of Allergic Rhinitis And Asthma

ObjectiveThe Toll-like receptor 9(TLR9)expressed by neutrophils and monocytes was used as the research object.Through constructing wild type(WT)mouse and FcεR Ⅰ gene knockout(FcεR Ⅰ KO)mouse Animal models of allergic rhinitis(AR)and allergic asthma(AA)and collectting specimens.in addition,collectting Peripheral blood of volunteers(including healthy people,AR,AA,and allergic rhinitis with asthma(ARA)).The expression of TLR9 in neutrophils and monocytes were detected by flow cytometry.To explore the effect of TLR9 expression in these two kinds of cells on the disease,and to study the effect of FcεRⅠ on the expression of TLR9.To reveal the role of TLR9 in the pathogenesis of AR,AA and ARA.Methods1.Human peripheral blood experiment: In this study,18 healthy volunteers were recruited as healthy control(HC),36 patients with AR,24 patients with AA and 21 patients with ARA,and peripheral blood of the volunteers was collected.The allergens(Platanus pollen allergen extract(PPE),House dust mite allergen extract,House Dust mite allergen extract,were detected by flow cytometry.HDME),Artemisia sieversiana wild allergen extract(ASWE))and the changes of TLR9 expression in human peripheral blood neutrophils and monocytes before and after stimulation.2.Animal experiments: AR and AA models were constructed in WT mice and FcεRⅠ KO mice by HDME and ASWE,and the expression changes of TLR9 in neutrophils and monocytes in blood of AR and AA mice and in neutrophils and macrophages in lung tissue of AA mice were detected by flow cytometry after specimen collection.To explore the role of TLR9 in AR and AA airway inflammation.Results1.Flow cytometry was used to detect TLR9 expression changes in peripheral blood neutrophils and monocytes of AR,AA,ARA patients and healthy controls before and after allergen stimulation.Without allergen stimulation,the ratio of neutrophils to monocyte TLR9~+ cells in the AR,AA and ARA groups were significantly increased compared with the HC group,while the mean fluorescence Intensity(MFI)expressed in TLR9 in the two types of cells was also significantly upregated.In addition,the proportion of TLR9~+ cells in peripheral blood neutrophils and monocyte TLR9~+ cells in AR group were significantly increased after stimulation by allergen HDME,while MFI was significantly up-regulated,and the proportion of TLR9~+ neutrophils was increased after stimulation by allergen ASWE.The MFI of TLR9 expression of monocytes in AR patients was up-regulated after stimulation by HDME,and the proportion of TLR9~+ monocytes in AA patients was increased after stimulation by allergen ASWE.However,the ratio of TLR9~+neutrophils/monocytes and the corresponding MFI in HC group were not significantly changed after the allergen excitation.2.Flow cytometry was used to detect TLR9 expression in blood neutrophils and monocytes of AR and AA models.Compared with FcεRⅠ KO mouse,TLR9 expression was up-regulated in neutrophils and monocytes after sensitization of allergens ASWE and HDME in WT mouse,and TLR9 expression was up-regulated in neutrophils and monocytes after sensitization of allergens in WT mouse and FcεR ⅠKO mouse.3.Flow cytometry was used to detect the changes of TLR9 expression in neutrophils and macrophages in lung tissue of AA model mouse.Compared with FcεR Ⅰ KO mouse,TLR9 expression in neutrophils and macrophages in WT mouse was up-regulated after sensitizing to allergens ASWE and HDME except for the changes of MFI in neutrophils.In WT mice and FcεR Ⅰ KO mouse,TLR9 expression was up-regulated in neutrophils and monocytes after allergen sensitization.Conclusion1.TLR9 in neutrophils and Monocyte participates in the pathophysiological process of allergic airway disease,The body may affect the allergic process by regulating the expression of TLR9 in neutrophils and Monocyte.2.The expression of TLR9 in neutrophils and Monocyte/macrophages may be affected by FcεRⅠ receptors.