Intervention Of Trichinella Spiralis ES Antigen In Allergic Asthmatic Mice

Allergic asthma（AA）is a chronic airway inflammatory disease,which is mainly infiltrated by inflammatory cells such as EOS,mainly manifested as airway hyperresponsiveness,airway inflammation and increased airway mucus secretion.Studies have shown that worms have great potential in regulating the host immune response.It is essential to develop new methods for the treatment of allergic asthma by using the Excretion of secretory antigen（ES）or derived protein of worms to replace the in vivo treatment of allergic asthma.This study selected Trichinella spiralis（T.spiralis）ES P43,P49,P53 as the research object,and further analyzed its intervention effect and potential immunological mechanism on allergic asthma induced by Ovalbumin（OVA）,so as to lay the research foundation for the development of new and effective worm-derived proteins/drugs for the treatment of allergic asthma.In this study,36 6–8 week-old BALB/c female mice were all SPF.Randomly divided into 6groups,blank control group,allergic asthma group,P43 intervention group,P49 intervention group,P53 intervention group,combined intervention group（P43+P49+P53）.The body weight and behavioral changes of mice in each group after excitation were recorded every 7 days,and the physical condition of mice was evaluated.Bronchoalveolar lavage fluid（BALF）was collected and Wright-Giemsa staining was performed to analyze the number of inflammatory cell infiltration.Hematoxylin-Eosin staining（HE）and Periodic Acid-Schiff stain（PAS）were used to observe the infiltration of inflammatory cells and airway mucus secretion.The levels of OVA-Ig E,OVA-Ig G₁,OVA-Ig G_2a,IL-4,IL-10,IFN-γand TGF-βin serum and the levels of IL-4,IL-10,IFN-γand TGF-βin bronchoalveolar lavage fluid were detected by Enzyme Linked Immunosorbent Assay（ELISA）.The expression of inflammatory factors in the pathogenesis of asthma was analyzed.The expression levels of T-bet and GATA-3 were detected by fluorescence quantitative PCR（q PCR）,Western Blotting（WB）and Immunohistochemistry（IHC）,and the direction of Th1/Th2immune differentiation was analyzed.Flow Cyto Metry（FCM）was used to analyze the differentiation of CD4⁺and CD8⁺cells in spleen single cell suspension of mice in each group during the pathogenesis of asthma.After P43,P49,P53 single intervention and combined intervention,the body weight of mice decreased slowly after excitation.During the behavioral score of mice in each group,there were occasional symptoms such as sneezing and flat breathing.The number of inflammatory cells in the lungs of mice decreased significantly.There were only a small amount of inflammatory cells around the trachea.The basement membrane was thin.A small amount of darkly stained goblet cell hyperplasia was observed in the bronchial wall.There was micro mucus secretion.The area of goblet cell mucus secretion decreased significantly.The contents of IL-4,OVA-Ig E and OVA-Ig G₁in serum and BALF of asthmatic mice decreased significantly.The levels of IFN-γ,TGF-β,IL-10,OVA-Ig G_2awere significantly increased,and the effect of combined intervention group was better than that of single intervention group.WB and q PCR results showed that after P43,P49,P53single intervention and combined intervention,the expression of T-bet in asthma group was increased but lower than that in control group,and the expression of GATA-3 was decreased but higher than that in control group.IHC also showed that the expression intensity and expression range of T-bet were significantly increased,and the expression intensity and expression range of GATA-3 were decreased.Overall,the effect of combined intervention group was better than that of single intervention group in the pathogenesis of asthma.The results of FCM showed that the ratio of CD4⁺/CD8⁺T cells in the asthma group was significantly higher than that in the control group,and the ratio of CD4⁺/CD8⁺T cells was significantly decreased after P43,P49 and P53 alone and combined intervention.In conclusion,this study showed that recombinant T.spiralis ES had a certain intervention effect on allergic asthma.T.spiralis ES antigen could reduce the infiltration of inflammatory cells and the secretion of airway mucus,regulate related cytokines and transcription factors,reverse the imbalance of Th1/Th2 cells and regulate the differentiation direction of CD4⁺/CD8⁺cells,and participate in inhibiting the inflammatory symptoms of allergic asthma mice.Therefore,the results of this study can provide a theoretical basis for the development of new and effective worm-derived drugs.