Expression And Significance Of TFH Cells And Its Related Factorsin Mouse Model With Allergic Rhinitis

Part I Establishment of mouse model with allergic rhinitisObjective:To establish a mouse model with allergic rhinitis, explore tissue changes in nasal mucosa with challenge times.Methods:BALB/c mice,4to6weeks old with50μg ovalbumin, aluminium hydroxide gel5mg and lml saline sensitized by intraperitoneal injection of7times;5%ovalbumin solution dropped for nasal provocation, were continuous applied of10days,20days,30days. The control group was replaced by0.9%saline. To observe the mice behavior, the level of OVA-sIgE by ELISA, nasal mucosa infiltration of eosinophils by HE, the results was analyzed using SPSS16.0.Results:The mice behavior scores of experiment were5.01±1.01,6.43±1.12,8.56±1.64, control group0.85±0.23; nasal mucosal EOS counts were5.63±1.25,9.25±2.10,15.37±2.36,0.69±0.12in the control group; the serum OVA-sIgE concentration was45.91(45.91±5.80)pg/ml、48.81(48.81±3.49) pg/ml、51.38(51.38±3.19) pg/ml, in control group (41.71±2.58pg/ml). T test showed that there were significant differences between the experimental group and the control (P<0.05), and between the model groups (P<0.05).Conclusion:Allergic rhinitis mouse model was established successfully; more nasal challenges, the concentration of OVA-sIgE and the symptoms were more increased. Part II Flow cytometric analysis of TFH cells in allergic rhinitis mouse modelObjective:To investigate the expression and significance of TFH cells in allergic rhinitis mouse model.Methods:After allergic rhinitis mice model established successfully,peripheral blood mononuclear cells(PBMC) were obtained with red cell lysates which were analyzed using CD4-Fitc and CXCR5-PE by flow cytometry to get the proportion of CD4+CXCR.5+cells in peripheral blood, the total serum OVA-sIgE levels was tested by ELISA, the relationship was analyzed by Pearson correlation in SPSS16.0.Results:TFH cells in AR model groups were respectively7.94±2.66%、8.65±3.49%、9.77±3.24%, the control group was4.97±0.20%, the difference was significant analyzed by T-test(P<0.05);differences between the model groups showed to be significant (T-test, P<0.05) and Pearson correlation analysis showed that the expression level of TFH cells and total OVA sIgE levels were positive (r=0.87, P=0.021)Conclusion:TFH cells have an important role in the pathogenesis of allergic rhinitis, which might be associated with the elevated OVA-sIgE. Part III Relationships between IL-21and TFH in mouse model with allergic rhinitisObjective:To investigate the expression of IL-21and its relationships between OVA-sIgE and TFH cells in allergic rhinitis mouse model. Methods:Peripheral blood was collected from allergic rhinitis BALB/c mice to test IL-21and OVA-sIgE by ELISA quantity, the percentage of TFH cells in PBMC by flow cytometry. its relationships between them were analyzed by Pearson correlation in SPSS16.0.Results:The expressions of11-21were186.25±11.68pg/ml in the control,181.13±4.93pg/ml in group AR1,168.46±11.04pg/ml in group AR2,164.54±10.52pg/ml in group AR3. After statistical analysis, IL-21showed to be negative correction with CD4+CXCR5+T cell ratio((r=-0.80, P=0.048) and total serum OVA-sIgE levels (r=-0.86, P=0.031)Conclusion:IL-21was negative correction with CD4+CXCR5+T cell ratio and total serum OVA-sIgE levels; it might have some therapeutic effect on allergic rhinitis in clinic. Part IV Expression and significance of TFH related cytokines in mouse nasal mucosa with allergic rhinitisObjective:To investigate the expression and significance of TFH related factor CXCR5and Bcl-6in allergic rhinitis of mouse nasal mucosa.Methods:The expression of CXCR5in nasal mucosa was analysed by immunohistochemical method. The TFH transcription factor of Bcl-6was tested by means of Westernblot and PCR, data processing and compare were used by medical statistical software SPSS16.0.Results:Westernblot showed the expression of bcl-6factor was increased significantly in the experimental groups, the relative density values were0.67,0.84,0.98, while the control was0.53. PCR assay showed Bcl-6mRNA expression were significantly increased, the amplification were0.90,1.63,1.77,1.85,1.92individually (T-test, P<0.05). CXCR5+cells increased significantly from the control group to different experimental groups (T-test. P<0.05), and were widely distributed in the mucosa and submucosa.Conclussion:CXCR5+cells and Bcl-6expression increased in mice nasal mucosa,which might play a role in the pathogenesis of allergic rhinitis.