| Background:Follicular helper T(Tfh) cells and T helper9cells(Th9) are two newly discovered typesof new CD4~+cell groups which are different from Th1,Th2,Th17and Treg cells.Themost significant characters of Tfh cells are their membrane surface expression markersCXC Chemokine Receptor5(CXCR5) and Inducible co-stimulator(ICOS) which cancombine with B cell—attracting chemokine-1(BCA-1/CXCL13) and Inducibleco-stimulator ligand(ICOSL） respectively,and secrete Interleukin21(IL-21).Tfh cellscan be migrated to the germinal centers of secondary lymphoid follicles (GerminalCenter, GC), stimulate their initial B cells activating to plasma cells with function of Igsecreting, and then help B cells produce abnormally high affinity antibody. Thecharacter of Th9can secrete Interferon9,which could increase the number ofinflammatory cells and promote the reaction of inflammation[5-7].Rheumatoid Arthritis（RA） is an autoimmune disease which mostly manifests joint swollen andpainful.Patients with RA exist varieties of abnormous autoimmune autibodies andinflammation factors in vivo.we conjectured Tfh and Th9cells maybe play a role inmechanisms of RA,analysing the relation among those data,such asautoantibodies,Inflammation factors and clinic data,detecting the relation withdisorders.Objective:To investigate the difference of Tfh and Th9cells by detecting the expression ofCXCR5~+CD4~+ICOS~+T(Tfh), CD3~+CD8-IL9~+（Th9），IL-21，CXCL13，IL-9in RApatients and normol controls. To analyse the probablely relationship between the Tfh, Th9，IL-21，CXCL13，IL-9and the disease duration,number of tender joints, number ofswollen joints,28-joint Disease Activity Score (DAS28), erythrocyte sedimentation rate(ESR), C-reactive protein (CRP), Anti-Cyclic Citrullinated Peptide antibody(Anti-CCPautibody) and Rheumatoid factor(RF), bone ersion.Methods：77cases of RA patients who come from the Rheumatology Department of the FirstAffiliated Hospital of Anhui Medical University and51cases of age-and gender-matched health controls from Healthy Check centre were collected during January toApril in the year of2011.All RA patients fited the1987(American College ofRheumatology， ACR)[8]. According to DAS28, patients were divided into low（2.6<DAS28≤3.2, n=15）,moderate（3.2<DAS28≤5.1,n=31）and high active group（DAS28>5.1,n=31）. Those patients included8who had never received any drugs,21with were in unofficial treatment and48were in standard treatment.We acquriedperipheral blood mononuclear cells(PBMCs) from patients and normol controls,detecting the expression of Tfh in peripheral blood(PB) which marked by CD4-FITC，CXCR5-PE，ICOS-APC through Flow Cytometry（FCM）. Investigated the levels ofIl-21,CXCL13in plasma of RA patients by enzyme-linked immunosorbent assay(ELISA). And analysed the correlation between percentage of Tfh cells in patients andclinic data,such as ESR, CRP, DAS28,RF,TJC, SJC and morning stiffness, etc..36casesof RA patients who come from the Rheumatology Department of the First AffiliatedHospital of Anhui Medical University and22cases of age-and gender-matched healthcontrols from Healthy Check centre were collected during May to July in the year of2011.According to DAS28,patients were divided into moderate（3.2<DAS28≤5.1,n=14）and high active group（DAS28<5.1,n=22）.We acquried PBMCs from patients andnormol controls,detecting the expression of Th9in PB which marked by CD8-FITC，IL9-PE，CD3-APC through FCM.Investigated the levels of Il-9in plasma of RApatients by ELISA. Results：1.The expression of Tfh cells in RA: The percentage of Tfh in patients were significanthigher than normol contro（l2.99±1.24VS1.07±0.42，P<0.001）.However, the levels ofIL-21（1.668±0.392VS1.264±0.374,P<0.001）and CXCL13（1.499±0.470VS0.999±0.165,P<0.001）were also higher than normol control.2.The percentage of Th9in patients were significant higher than normol control（0.989±0.498VS0.213±0.084，P<0.001）.The levels of IL-9were also higher thannormol control（1.055±0.421VS0.686±0.134，p<0.01）3. The expression of Tfh（1.85±0.66,2.46±0.64,4.02±1.16） and the levels of IL-21（1.435±0.303vs1.638±0.330vs1.908±0.415）and CXCL13（1.179±0.245vs1.434±0.431vs1.756±0.527） were increased markedly in low, moderate and highdisease activity groups.(P <0.01). The following of those data in three disease activitygroups were statisticsly signficance（F=8.075，6.125，5.364，P<0.001）.4.However,The expression of Th9（0.686±0.254vs1.181±0.523）and the levels of IL-9（0.939±0.301vs1.367±0.470）were increased markedly in moderate and high diseaseactivity groups.Those percentage were also higher than NC(P <0.01).The following ofthose data in two disease activity groups were statisticsly signficance between each twogroups.4.The expression of Tfh in those patients who have bone destruction were higher thanno or mild bone destruction（3.36±1.28VS2.67±1.11,P <0.05）.5.There was a positive correlation with Tfh and DAS28(r=0.352, p=0.002),ESR(r=0.230,p=0.004),TJC(r=0.269,p=0.018),bone erosion（r=0.306,P=0.007,RF(r=0.622,p=0.000),Anti-CCP(r=0.283,p=0.026),IL-21(r=0.367,p=0.001),and CXCL13(r=0.283,p=0.026)respectively.There were no relationship with Tfh and course，morning stiffness andSJC.And plasma IL-21had conversation with DAS (r=0.469,p=0.000), ESR(r=0.387,p=0.002),RF(r=0.306,p=0.0015),TJC(r=0.344,p=0.006),CXCL13(r=0.435,p=0.000).Besides,CXCL13was correlation with DAS(r=0.415,p=0.001), ESR(r=0.410,p=0.001), CRP(r=0.353,p=0.005),TJC(r=0.252,p=0.046).6.There was a positive correlation with Th9and TJC(r=0.341,p=0.042), SJC(r=0.347,p=0.038),ESR(r=0.347,p=0.042),CRP(r=0.384,p=0.021),DAS28(r=0.461,p=0.005) andRF(r=0.379,p=0.025).There were no relationship with Th9and course，morningstiffness, Anti-CCP and bone ersion.There was a positive correlation with IL-9andTJC(r=0.420,p=0.023),SJC(r=0.361,p=0.035),ESR(r=0.403,p=0.018)and DAS28(r=0.385,p=0.048).There were no relationship with IL-9and course，morning stiffness,CRP, Anti-CCP,RF, and bone ersion.7.The expression of Tfh in uncured RA patients，unnormal cured RA patients，curedRA patients were decrease in turn (3.43±1.26vs2.83±1.26vs2.73±0.85，P<0.001)；What’s more，the expression of Tfh in cured RA patients were lower than uncuredRA patients.Conclusion:1.The expression of Tfh cells,IL-21and CXCL13in RA patients increased markedlythan that in health controls, fourthmore, its enhanced as much as the strenghthen diseaseacyivity.Tfh cells were postive relationship with the DAS28,TJC,ESR,CRP and boneersion.These results described that the abnormality of Tfh may play an important rolein the pathogenesis of RA and be an indicatior of disease activity.2.Tfh cells are closely related to the autoimmune antibodies,such as Anti-CCP,RF. Theresults may show that Tfh cells could assist B cells in generating abnormol high affinityantibody.3.The expression of Tfh were decrease by curing, which may be applied to a results thatthe treatment of RA patients through monitor Tfh.4.The expression of Th9cells,IL9in RA patients increased markedly than that inhealth controls, and Th9cells were postive relationship with the DAS28,TJC,ESR,CRP.These results described that the close relation between Th9and inflammation,whichmight be one of the target to promote the disorders developing. |
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