The Comparision Of Immunological Changes Between Bronchiolitis And Asthma

PART I COMPARISONS OF CD4+ T LYMPHOCYTE SUBGROUP IN PERIPHERAL BLOOD MONONUCLEAR CELLS(PBMCS) BETWEEN THE BRONCHIOLITIS IN THE ACUTE PHASE AND ASTHMA ACUTE ATTACK PATIENTS.Objective: To understand the differences of the proportion of Th1,Th2,Th17 and Treg cells/CD4 T lymphacytes in PBMCs and their cytokines in serum between the bronchiolitis in the acute phase and asthma acute attack patients.Method:31 children with bronchiolitis and 26 patients with asthma acute attack,and 18 healthy children were included.The proportion of Th1,Th2,Th17,and Treg cells/CD4 in PBMCs were analyzed with flow cytometry. Serum IL-4,IL-10,IL-17和IFN-γwere assayed by ELISA.Result:The proportion of blood Th1 cell of bronchiolitis patients was lower than that of the controls and asthma (P=0.000 and P=0.010 respectively). The proportion of blood Th1 cell of asthma have decrease tendency than that of the controls, but have no statistical significance (P=0.096). The proportion of Th2 cells have increase tendency than that of the controls, but have no statistical significance (P=0.357) and was lower than that of the asthma (P=0.006),meanwhile the proportion of Th2 cells in asthma group was higher than that of the control group (P=0.001). The proportion of blood Th17 cell have increase tendency than that of the controls, but have no statistical significance (P=0.336) and have decrease tendency than that of the control, but have no statistical significance (P=0.333). The proportion of blood Treg cell of bronchiolitis patients was higher than that of the controls and asthma (P=0.000 and P=0.010 respectively). The Th1/Th2 ratio analysis showed that the Th1/Th2 ratio in the control group was 10.22±9.51,in asthma group was 5.12±2.19,in bronchiolitis group is 5.52±1.16. The Th1/Th2 ratio of bronchiolitis group when comparing with that of asthma group (P =0.763),when comparing with that of the controls (P =0.000).The serum of IL-4 of bronchiolitis group have increase tendency than that of the controls, but have no statistical significance (P=0.893) and have decrease tendency than that of the control, but have no statistical significance (P=0.096). The serum of IL-17 of bronchiolitis group have increase tendency than that of the control, but have no statistical significance (P=0.451) and have decrease tendency than that of the control, but have no statistical significance (P=0.363). The serum IL-10 was lower than that of the control and asthma group(P=0.002 and P=0.023 respectively).Conclusions: Th bronchiolitis have decreased Th1 cells subgroup, unapparently increased Th2 cells subgroup, and apparently decreased Treg cells subgroup compared with that of the controls;The asthma have decreased Th1 cells and Treg cells subgroup, but increased Th2 cells subgroup apparently compared with that of the controls. The Th1/Th2 ratio may have parallel decreasing imbalance tendency in these two patients group. And the proportion of Treg subgroup of the bronchiolitis group increased but may be insufficiency of function.PART II THE INFLUENCE OF REGULATORY T LYMPHOCYTE IN PERIPHERAL BLOOD MONONUCLEAR CELLS OF THE BRONCHIOLITIS IN ACUTE PHASE ON THE POST-BRONCHIOLITIS WHEEZINGObjective: To understand the influence of regulatory T lymphocyte,and other T cells subgroup including Th1,Th2,Th17 cells in peripheral blood mononuclear cells( PBMCs) of the bronchiolitis in acute phase on the post-bronchiolitis wheezing.Method:The post-bronchiolitis wheezing was followed up by telephones in recent one year. 11 children without wheezing and 7 patients with wheezing were included.The proportion of Th1,Th2,Th17,and Treg cells/CD4+ T lymphacytes in PBMCs were analyzed with flow cytometry. Serum IL-4,IL-10,IL-17 and IFN-γwere assayed by ELISA.Result:The comparisons of the proportion of Thl,Th2,Thl7,Treg cells/CD4+T lymphocyte in PBMCs of the bronchiolitis in acute phase showed that the post-bronchiolitis wheezing group have increase tendency of proportion of Th17 cells subgroup and decrease tendency of proportion of Th1 and Th2 cells subgroup than that of the non-wheezing group, but none of them have statistical significances. And the proportion of Treg cells was lower than that of the non-wheezing group(P=0.023).The level of serum IL- 17,IL- 4,IL-10 and IFN-γin the ? post-bronchiolitis wheezing group have increase tendency than that of non-wheezing group, but none of them have statistical significances.Conclusions:The development of post-bronchiolitis wheezing may have associations with the deficiency function of regulatory T lymphocytes in bronchiolitis in acute phase. PART III ASSOCIATIONS OF MINA GENE POLYMORPHISMS WITH CHILDREN ATOPIC WHEEZING IN A CHONGQING HAN POPULATIONObjective: To study the associations of the single-nucleotide polymorphisms polymorphisms(SNPs)of Mina gene rs832081(C/T),rs832078(C/T), rs4857304(G/T), rs9879532 (C/T), rs17374916(A/G), rs1532206(A/G) with children atopic wheezing in Chongqing Han people.Methods: The TAGGER program available in Haploview version 4.0 software was used to select the Tag polymorphisms. A case-control study was applied to observe the distributions of SNPs of Mina gene in the wheezing patients and controls. 202 cases of asthmatic children,aged 6.97±2.85 years,and 191 controls was included. SNP genotyping was performed by means of matrix assisted laser desorption ionisation-time of flight mass spectrometry method(MALDI-TOF).Result: Among the six SNPs, only rs4857304 of Mina gene of wheezing patients presented with a significantly higher portion of T allele frequencies(P=0.0199 vs control, OR=1.477, 95%CI=1.062-2.052), but no statistical differences among the genotypes frequencies of rs4857304 compared with controls (P=0.0587 vs controls).Conclusions:Our results highlight the role of Mina as a susceptibility gene for children atopic wheezing in Chongqing Han people.PART IV MINA GENE RS4857304 POLYMORPHISMS CAN NOT JUDGE THE PRESCHOOL RECURRENT WHEEZING FROM THE ASTHMAObjective: To study the distribution differences of the polymorphisms of Mina gene rs4857304(G/T) between recurrent wheezing children and asthma children.Methods: 202 cases of asthmatic children,aged 6.97±2.85 years,and 191 controls was included. SNP genotyping was performed by means of matrix assisted laser desorption ionisation-time of flight mass spectrometry method(MALDI-TOF). The recessive genetic model analysis was used to study the differences of rs4857304 (G/T) genotype TT between TT+TG. The dominant genetic model analysis was used to study the differences of rs4857304 (G/T) genotype TT+TG between GG. The distribution differences of the polymorphisms of Mina gene rs4857304(G/T) between recurrent wheezing children and asthma children were studied by chi-square statistics. Result: By the recessive genetic model analysis found that the proportion of the main allele homozygous TT of rs4857304 was higher than the proportion of GT+TT type (P=0.0282, OR=1.568 ,95%CI=1.048-2.346). Those children were divide into recurrent wheezing and asthma children,so the recurrent wheezin group altogether had 58 cases,among them TT genotype were 39, and GT+GG genotype were 19; the asthma group altogether had 141 cases,among them TT genotype were 84, and GT+GG genotype were 57. Through chi-square statistics, the comparisons of TT genotype and GT+GG genotype had no statistical significances between recurrent wheezing and asthma children.Conclusions:Mina gene rs4857304 polymorphisms can not be used as one of the biomarker to judge the preschool recurrent wheezing from asthma.