Clinical And Basic Research On Eosinophils In Inflammatory Diseases Of Upper And Lower Airways

Part I: Clinical study of eosinophils in the upper and lowerairways of patients with chronic rhinitisSection One:The standardization of inflammation detecting methods in upper and lower airways Objective:To investigate the standardization of inflammation detecting methods in upper and lower airways.Method: After a five year cooperation with State Key Laboratory of Respiratory Diseases on inflammatory diseases of airways,we have found a series of evaluation methodology and normative values in upper and lower airways(nitric oxide(NO),Airway Hyper Reactivity and cytology(EOS)).Result:The normative range of nasal and pulmonary NO is 400~900ppb and 5~25ppb respectively.The nasal resistance increased ?100% and FEV1 fell ?20%when compared with their respective baselines-both illustrating a positive result.The positive value of nasal and pulmonary EOS are ?2.00/HP and ? 2.5%respectively.Conclusion: The standardization of evaluation methods for upper and lower airway inflammation provides the methodology and research basis for follow-up studies of upper and lower airways.Section Two:The significance of eosinophils in thecorrelation of upper and lower airway inflammation inpatients with chronic rhinitis Objective: To explore the index of nasal inflammation predicting lower airway inflammation by investigating the expression and association of EOS in the upper-lower airways and blood of patients with chronic rhinitis.Methods: A total of 162 patients with allergic rhinitis(AR),117 patients with non-allergic rhinitis(NAR)and 104 controls(Ctrl)were enrolled.All subjects detailed medical history collection,skin prick test(SPT),blood test,nasal lavage and induced sputum EOS,nasal provocation and bronchial provocation test(NPT,BPT),nasal and forced exhaled nitric oxide(NNO,FeNO).One-way analysis of variance was used for comparison between groups.LSD t test or Mann-Whitney U test was used for comparison between the two groups.Correlation analysis using Pearson or Spearman related parameter test.Results: There were significant differences in the inflammatory indexes between the AR and NAR groups compared with the Ctrl group(p<0.01).The nasal lavage EOS,NNO,induced sputum EOS,FeNO,blood EOS and tIgE were higher in the AR group than in the NAR group(p<0.01),but there was no significant difference in nasal resistance,NPT and BPT between the two groups.Nasal lavage EOS in AR group,NAR group,or rhinitis group(AR+NAR)was correlated with induced sputum EOS,FeNO,tIgE,and blood EOS(p<0.05),but not with BPT.After adjustment for gender,age,height and weight,nasal EOS was positively correlated with sputum EOS(p<0.01).Multiple linear regression analysis showed that nasal EOS,blood EOS and SPT were factors affecting sputum EOS levels(p<0.05).Logistic regression analysis of induced sputum EOS was found to be meaningful in the AR group,NAR group and rhinitis group.The correct classification ability was77.2%,79.5% and 78.1%,respectively(P< 0.001),while FeNO and BPT did not find a valuable indicator of the nose.The optimal threshold for nasal EOS to determine induced sputum EOS is 3.30/200 HP by ROC analysis.Conclusion: 1.The inflammatory index of the AR group is heavier than that of the NAR group,but significantly higher than the control group.2.The nasal EOS is correlated with multiple lower airway and systemic inflammatory markers,and is a risk factor for the induced sputum EOS.Part II: The establishment of a model of allergic airwayinflammation with consistent airway in the upper andlower airways of mice Objective: Establish an animal model of allergic airway inflammation with consistent airway in the upper and lower airways of mice Method: OVA-sensitized BALB/c mice were intranasal OVA exposure three times a week for 3 weeks,followed by 5 days of OVA aerosol challenge,and the symptoms,serum sIgE concentration and pathological changes of nasal lung mucosa were evaluated in the mouse model after the last challenge.Result: The nasal and pulmonary symptom scores of the model group were significantly higher than those of the ctrl group(P(27)0.01).There was a statistically significant difference of nasal and pulmonary mucosal damage(grades 1,2)of the model group compared with the ctrl group(P(27)0.01),but there was no statistical significance at grade 3(P(29)0.05).Nasal lavage,lung lavage and serum OVAsIgE concentrations in the model group were significantly higher than those in the ctrl group(P(27)0.01).The nasal and pulmonary EOS and GC in the model group were significantly higher than those in the ctrl group(P(27)0.01).There was a significant positive correlation between nasal and pulmonary EOS in the model group(r=0.775,P(27)0.01),and the number of GC was also positively correlated(r=0.723,P(27)0.05).Conclusion: The modeling method can successfully establish AAI model with upper and lower airway consistency in three aspects of symptom,immunology and pathology in mice.Part III: Effects of nasal administration on the upper andlower airways in mice with allergic airway inflammationand its mechanism Objective: AAI mice were treated with HMGB1 and its receptor blocker(RAGE)to observe its therapeutic effect and to explain its possible mechanism.Method: 32 BALB/c mice were randomly divided into model group(A),HMGB1group(B),RAGE group(C)and control group(D).OVA-sensitized BALB/c mice were intranasal OVA exposure three times a week for 3 weeks.HMGB1 was administered to a subset of these mice 1 hour after intranasal OVA challenge,but RAGE 1 hours before,followed by 5 days of OVA aerosol challenge of group A?B?C.The assessment of nasal lung symptoms,serum cytokines(OVAsIgE,IL4,IL10,IL17,and IFN-?)and pathological changes(EOS,GC)after the last challenge.Result: The scores of nasal symptoms in group A,B and C were significantly higher than those in group D(P(27)0.01),and group C was significantly better than group A ?B(P(27)0.01).The pulmonary symptom scores of group A,B and C were significantly different from those of group D(P(27)0.01),There was significant difference between group C and group A ? B(P(27)0.01),There was no statistical difference between group A and B(P(29)0.05).The concentrations of serum OVAsIgE in group A,B and C were significantly higher than those in the group D(P(27)0.01).Group A and B were also significantly higher than group C(P(27)0.01).There was no significant difference between group A and B(P(29)0.05).IL-4 A,B,C were higher in the three groups than in the group D(P(27)0.01),and group C was lower than the group A,B(P(27)0.01),and there was no significantdifference between group A,B(P(29)0.05).IL-10 A,B,C were higher in the three groups than in the group D(P(27)0.01),and the group C was lower than the group A,B(P(27)0.01),and there was a statistically significant difference between group A,B(P(27)0.05).IL-17 A,B,and C groups were higher than the group D(P(27)0.01),and there were significant statistical differences between the AC,BC,and AB groups(P(27)0.01).IFN-? there were significant differences between the AC,BC,AD and BD groups(P(27)0.01),but no significant differences between the AB and CD groups(P(29)0.05).The ratios of IFN-?/IL4 in group A,B and C were lower than those in the group D(P(27)0.01),and those in group C were higher than those in group A and B(P(27)0.01).There was no significant difference between AB(P(29)0.05).There was no significant difference in the ratio of IL-17/IL-10 among A,B,C and D groups(P(29)0.05).The levels of EOS infiltration and 1000 um long GC in the nasal and pulmonary mucosa of group A,B and C were significantly higher than those in the group D(P(27)0.01).The four indexes of nasal,pulmonary EOS and GC between AC and BC were significant difference(P(27)0.01),but there was no statistical difference in the AB group(P(29)0.05).There was a positive correlation between the number of EOS in the nasal and pulmonary tissues of group B(r=0.841;p=0.009),and the number of GC was also positively correlated(r=0.781;p=0.022).The number of EOS in nasal and pulmonary mucosa of group C was also positively correlated(r= 0.744;p=0.034),while there was no significant correlation of GC(P(29)0.05).Conclusion: 1.This study confirmed that HMGB1 can aggravate the level of partial inflammation in AAI mice,and RAGE can significantly reduce the level ofAAI inflammation;2.The involvement of HMGB1 in the regulation of AAI may be achieved by reducing the tissue EOS infiltration after changing the Th1/Th2 balance;3.The nasal EOS is positively correlated with the lung EOS,but the GC correlation of the nasal lung is not as good as EOS.