哮喘小鼠骨髓树突状细胞的表型及共刺激分子研究.doc

DCs rmGMCSFrmIL-4DCsDCsDCsT DCsDCs(33D1)DCsCD86CD40CD80DCsTDCs DCsDCs B7CD40 The study of phenotypes and costimulatory molecules of dendritic cells in asthmatic mice WU Jian-qingSUN YunYIN Kai-sheng.The First Affiliated Hospital of Nanjing Medical UniversityJiangsu 210029China Abstract Objective The purpose of this study was to evaluate the effects of dendritic cells derived from bone marrow in the pathogenesis of asthma.Methods Murine model of ovalbumin (OVA)allergic asthma was established.DCs were developed by incubating bone marrow cells with rmIL-4 and granulocyte-macrophage colonystimulating factor. Phenotype was assessed with flow cytometryand the antigenpresenting function was assessed with the mixed leukocyte reaction.Results DCs expressed DCs characteristic 33D1 can be developed from bone marrow cells by using rmIL-4 and rmGMCSF.The stimulating effect of the DCs on the proliferation of the lymphocytes was shown in asthmatic and control groupbut the effect in asthmatic group was more intensive than control.DCs from asthmatic mice showed phenotypic differences in the expression of CD86but there were no differences in the expression of CD80 and CD40 between asthmatic and controlled group.Conclusion This study suggests the antigenpresenting function of DCs in asthmatic group was increased and DCs may play an important role in the pathogenesis of asthma. Key words asthma;dendritic cells;B7;CD40;mixed leukocyte reaction (Eos)T(AHR)CD4+TTh2B7.1(CD80)B7.2CD8612dendritic cellsDCsantigen-presenting cellsAPCsTAPCsDCsAPCsT!-empirenews.page- 1 1.1 BALB/C46ICR68 1.2 -rmGM-CSF-4rmIL-4RD systemFITC antiMouse CD86FITC antiMouse CD80FITC antiMouse CD40AntiMouse DC Marker 33D1eBioscienceFITC Goat antiRat IgGH+L 1.3 OVAGradeSigmaRPMI1640(L-NaHCO3)GIBCO(FCS)HycloneCKaowa Hakko Kogyo(ConA)SigmaCellTiter 96kAQueousOne Solution Cell Proliferation Assay(MTS)Promega 1.4 1.4.1 3201018OVA0.1mlOVA 10mgAl(OH)3 20mg141%OVAS-888E30min2 1.4.2 DCs 4BALB/CHanks0.83%Tris-NH4Cl1106/ml241ml375%CO24hrmGMCSF1000u/mlrmIL-4500u/ml37 1.4.3 1.4.4 DCs 7DCs2105/ml1l FITCCD86CD80CD40DCs33D1430minDCs33D12l FITCIgG(H+L)430min 1.4.5 DCs25g/mlC375%CO24104/100l2104/100l1104/100l4103/100l963ICRT2105/100l200lConAT375%CO296h4h20l MTS4h490nmSISI=OD-OD/TOD-OD3!-empirenews.page- 1.5 xsSPSS10.0tP0.05 2 2.1 DCs rmGM-CSFrmIL-4232p13387DCDC 2.2 FACSDCsDCCD80CD40CD86DC33D14CD8651.608.7336.113.51P0.05CD15DC33123 D1IL-4DCs1 DC22 DC4 3 DC71 DC xs%n=33*P0.054 022043027024DC017038037019DC!-empirenews.page-5 DC2.3 DCT DCDCTDCDCDCTDC26DCTDCT7DCTP0.052 DCT xsn=3P0.001P0.01*P0.056 DCT7 DCT P0.01*P0.05 3 DC0.5%1.0%DCsInaba5rmGM-CSFDCDCDCDCGM-CSFGM-CSFDCDCIL-4GM-CSFDCIL-4DCDCDCT3MHCCD40CD86CD80ICAMTDC6DC733D1NLDC145OX62DCCD1aCD83DC33D1DCCD40CD80CD86DCT3DCs!-empirenews.page- & nbsp; DCTDC-MHCDCCD40CD86CD80TCD28CD80/CD86-CD28TTDCT8 CD4+TTh2DCTTh29ThTh2TCD80/CD86-CD28TCD80CD86CD28DCTh210CD80/CD86CD28Th211IL-5IL-412TIL-5CD8613CD86CD86TThTh2IL-5EosDCCD86P0.05DCCD86Th