| Background:From now on, although the molecular mechanisms underlying IPF pathogenesis are still elusive and controversial, current research suggests that abnormal and deregulated wound healing play the decisive role in the pathogenesis of the disease, while during the process of healing, inflammatory reaction occupies the significance position. T helper cells are the chief component of inflammatory cell and disequilibrium of Thl/Th2 made the important effect in fibrosis previously. Since Treg and Th17 have been discovered, they are considered to take part in the occurrence and development of IPF as Th1/Th2. Regarding Th1,Th2,Th17 and Treg cell differentiating from CD4+T cell, an extremely complicated network form while interaction and mutual inhibition among all the cells, the specific function of T helper lymphocyte is necessary to be proved by further research.ObjectiveTo investigate the effect and significance of Th1,Th2,Th17,Treg lymphocyte cell in IPF. The study detected the distribution and function of the four phenotype cells and the expression of transcription factor subtypes on peripheral blood lymphocytes, detected the response of subtypes of lymphocyte to PHA or rIL-2 stimulation by the level of subtypes of lymphocyte cytokines in blood serum and culture supernatant of PBMC, CD4+CD25-T cell, Treg, respectively.MethodsCollect the peripheral blood and BALF of the patients who diagnosed IPF and the control group; The peripheral blood mononuclear cells and the lymphocyte cells of the BLAF were cultured,stimulated, as well as in vitro markered and then detceted the percentage of the Thl phenotype cells, Th2 phenotye cells,Thl7 phenotype cells and Treg Phenotypes cell by flow cytometry, to analyse the relationship between the various subtypes of Th cell with IPF.The relevant levels of cytokines in peripheral blood were found through the enzyme linked immunosorbent assay to detect the relationship between function of the various subtypes of Th cell and the desease in IPF; Finally,The mRNA in peripheral blood mononuclear cells were extracted by Real-Time PCR to measure the expression of transcription factors of the four phenotypes cells. The PBMC, CD4+CD25-T cells, Treg cells were cultured were stimulated with PHA or rIL-2 in different incubation media (Fetal Bovine Serum culture and self-serum culture) respectively,and then the concentration of relative cytokines in superntant fluid of different incubation media were analyzed with ELISA to estimate the function of T helper cells in IPF furtherly.All data were expressed as Mean±standard deviation (x±s), The difference between different groups was assessed with Independent-Samples T test and Mann-whitney U test. Correlation analysis was assessed with Pearson and Spearman correlation coefficient test. If the P value was less than 0.05,It was considered to be a statistically significant difference.Results1. The percentage of Th1 cells in peripheral lymphocyte in IPF patients was lower than that in the control group, and there was statistical significance (P<0.05);The perceniage of Th2 cells in peripheral lymphocyte in IPF was higher than that in the control group, but there was no statistical significance (P>0.05); The percentage of Th17 cells in peripheral lymphocyte in IPF was higher than that in the control group, and there was statistical significance (P<0.05); The percentage of Treg cells in CD4+ lymphocyte in IPF was lower than that in the control, also there was statistical significance(P<0.05);2. The percentage of Treg cells in CD4+ lymphocyte in IPF was positively correlated with FVC pred%,TLC pred% and DLCO pred% respectively;3. The percentage of Treg cells in CD4+ lymphocyte in the BALF of IPF was lower than that in the control group, and there was statistical significance(P<0.05); the percentage of other subtypes of Th cells in peripheral lymphocyte in IPF were not different with that in the control;4. The expression of transcription factor of T-bet, GATA-3, FOXP3 lymphocytic mRNA of PBMC were correlated with the percentage of Th1,Th2 cells in PBMC in IPF, and with the percentage of Treg cells in CD4+lymphocyte in IPF. The expression of transcription factor of T-bet, FOXP3 lymphocytic mRNA of PBMC were positively correlated with the percentage of Th1 in PBMC and with the percentage of Treg cells in CD4+ lymphocyte in IPF respectively.5. The concentration of IFN-γ, IL-10 in blood serum of IPF patients were lower than that in the control group, and there were statistical significance (P<0.05); The concentration of IL-17A in blood serum of IPF patients were higher than that in the control group, also there were statistical significance (P<0.05); There was no obvious difference in the concentration of IL-4 in blood serum of two groups.6. The PBMCs and CD4+CD25-T cells were incubated in the different cultures, Fetal Bovine Serum culture(FBSc) and self-serum culture(SSc), the concentration of IFN-y, IL-17A were stimulated with PHA in superntant fluid of FBSc were higher than that in SSc. And there were statistical significance(P<0.05); the concentration of IL-4 in superntant fluid of FBSc was higher than that in SSc. but there was not statistical significance(P>0.05).7. The function of secreting-IL-10 of Treg cells were stimulated with rIL-2 in IPF patients was lower than that in the control group,and there were statistical significance(P<0.05).Conclusion:1. The T helper cells play an important role in the pathogenesis of IPF, Thl,Treg cells and IFN-y,IL-10 may have protected effect in the patients with IPF; Th2-type immune response may have no influence to IPF; Th17,IL-17A were took part in the pathogenesis of IPF.2. The expression of transcription factors determine the differentiation of subtypes of T helper cells in IPF patients.3. There was no obviously differences between the secreting-cytokine function itself of Th1,Th2 in the two groups, the difference of the concentration of IFN-y, IL-4,IL-17A in the different cultures indicate some inhibited factors in the self serum.4. The percentage of Treg cells was lower in the IPF patients that lead to the secreting-IL-10 function of Treg cells poorly. Our research render Tregs as candidate targets for an original therapy for the patients with IPF.
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