ABSTRACT
Objective To investigate the effects of tumor necrosis factor-related ligand-1A(TL1A)on activation of T helper 9(Th9)cells of colonic tissues in chronic experimental colitis mice.Methods The chronic experimental colitis mice model was established with drinking dextran sulfate sodium salt(DSS).A total of 32 lymphocytes TL1A highly expressed mice and wild type(WT)mice were divided into WT control group, transgene control group,WT modeling group and transgene modeling group.The mice of control groups were administrated with distilled water. The mice of modeling groups received 3% DSS in drinking water discontinuously.The mice were sacrificed on 29 days after modeling.Body mass was measured,length of colon was recorded,scores of gross colon and the disease activity index(DAI)were calculated.The colonic morphological changes were observed by hematoxylin-eosin(H-E)staining.The lamina propria mononuclear cells(LPMC)were isolated and the number of Th9 cells was tested by flow cytometry.The levels of interleukin-9(IL-9)in serum and LPMC were detected by enzyme-linked immunosorbent assay(ELISA).The expressions of IL-9 protein and mRNA of the colonic tissues were measured by Western blotting and real-time polymerase chain reaction(PCR),respectively.T test and single factor analysis of variance were performed for statistical analysis.Results The percentage of body mass loss of WT modeling group was lower than that of transgene modeling group(16.2% ± 1.0% vs 18.9% ± 1.2%),and the difference was statistically significant(t=4.90, P<0.05).The scores of gross colon,DAI and pathology of transgene modeling group were all higher than those of WT modeling group(2.80 ± 0.64 vs 1.60 ± 0.31,2.55 ± 0.20 vs 1.58 ± 0.17,and 11.85 ± 0.86 vs 9.50 ± 0.79),and the differences were statistically significant(t=4.77,10.45 and 5.69,all P<0.05).The number of LPMC in transgene modeling group was higher than that of WT modeling group(3.70×106± 0.28×106vs 2.65×106± 0.32 × 106)and the difference was statistically significant(t= 6.98,P< 0.05).The percentage of Th9 in total CD4+T cells of LPMC in colonic tissues of transgene modeling group was higher than that of WT modeling group(0.54% ± 0.04% vs 0.23% ± 0.03%),and the difference was statistically significant(t= 17.54,P< 0.05).The serum IL-9 level of transgene modeling group was higher than that of WT modeling group((170.23 ± 5.69)pg/mL vs(150.62 ± 6.45)pg/mL),and the difference was statistically significant(t= 6.50,P< 0.05).The level of IL-9 secreted by LMPC of transgene modeling group was higher than that of WT modeling group((265.21 ± 8.76)pg/mL vs (237.58 ± 10.24)pg/mL),and the difference was statistically significant(t= 5.80,P< 0.05).The expressions of IL-9 protein and mRNA of transgene modeling group were higher than those of WT modeling group(1.31 ± 0.09 vs 1.18 ± 0.03,and 8.26 ± 1.13 vs 2.25 ± 0.29,respectively),and the differences were statistically significant(t=3.88 and 14.57,both P< 0.05).Conclusion TL1A high expression in lymphocytes can promote Th9 cells differentiation and IL-9 secretion which involved in the genesis of chronic experimental colitis.
ABSTRACT
Objective To investgate the expression of Tfh and Th9 in the peripheral blood of rheumatoid arthritis ( RA),and to analyze the relationship between those cells and disease activity,organs of involvement,ect.,and to underly the possible immunological mechananism of RA.Methods Peripheral blood Tfh cells and Th9 cells,define as the CD4+ CXCR5 + ICOS+ and CD3 + CD8+ IL-9+ population,were examined by flow cytometry in 36 patients with RA and 22 case of age-and gender-matched health controls.According to 28-joint Disease Activity Score (DAS28),patients were divied into two groups:high disease activity(n=22),moderate disease activity(n=14).And then to analyzed the correlation among the Tfh,Th9 and clinical data of RA,such as ESR,CRP,RF,the number of tender joints,the number of swollen joints and bone destruction,etc..While to analyzed the correlation between Th9 and Tfh.Results The expression of Tfh (Z=-6.082,P=0.000) and Th9 (0.989±0.498 vs 0.213±0.084,t=13.063,P =0.000 )in PBMCs of RA was significantly higher than normal controls.Meanwhile,the expression of Tfh ( 3.880 ±1.255 vs 2.678±1.022,t=2.990,P=0.005 ) and Th9( 1.181 ±0.523 vs 0.686±0.254,t =4.043,P=0.000) in high activity of group were higher than those moderate.The percentage of Tfh and Th9 in the two groups were all signifieantly higher than controls(P＜0.01 ),There was a positive correlation with Tfh and DAS28 ( r =0.571,P=0.000),ESR ( r =0.375,P =0.029),CRP( r =0.357,P =0.032 ),the number of tender joints ( r=0.598,P =0.000 ),RF ( r =0.421,P =0.023 ) and anti-CCP ( r =0.421,P =0.023 ).There were no relationship with Tfh and course,morning stiffness,the number of swollen joints,bone erosion and the abnormal of EKG.There was a positive correlation with Th9 and DAS28( r=0.461,P=0.005 ),ESR(r=0.347,P=0.042),CRP(r=0.384,P=0.210),the number of tender joints(r=0.341,P=0.042),the number of swollen joints (r =0.347,P=0.038 ) and RF( r =0.379,P =0.025 ) respectively.There were no relationship with Th9 and course,morning stiffness,CCP,the abnormal of EKG,bone erosion.There was a positive correlation with Tfh and Th9 (r=0.727,P =0.000),Conclusion The expression of Tfh and Th9 were increase significantly,which those cells were close relationship with the disease activity and some data,These results indicate that the abnormality of Tfh and Th9 may play an important role in the pathogenesis of RA.