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机构地区:[1]安徽医科大学免疫学教研室及过敏与免疫研究中心,安徽合肥230032
出 处:《安徽医药》2015年第2期276-279,共4页Anhui Medical and Pharmaceutical Journal
基 金:国家自然科学基金(No 81273245);安徽省国际科技合作计划(No 11030603027)
摘 要:目的观察添加人AB型血清或胎牛血清(FBS)培养基培养的人外周血单个核细胞(PBMC)在结核杆菌(M.tb)全菌裂解物(WCL)的刺激下增殖差异。方法健康成人PBMC 1×109·L-1,分别加入WCL 20 mg·L-1,或者同时给予人重组白细胞介素-2(rh IL-2)50 U·m L-1,植物血凝素(PHA)5 mg·L-1,同时设置空白对照组。分别在含10%人AB型血清的RPMI1640培养液和10%FBS的RPMIl640培养液中培养6 d后,用倒置显微镜观察培养结果,运用流式细胞术检测淋巴细胞增殖情况。结果添加人AB型血清培养基培养条件下,M.tb WCL对淋巴细胞增殖效果较同样条件下添加FBS培养的增殖效果明显,差异有统计学意义(P<0.05)。在WCL和rh IL-2混合组中同样是添加人AB型血清培养增殖效果明显,差异有统计学意义(P<0.05)。结论当以M.tb WCL为抗原的刺激人PBMC时,淋巴细胞在添加人AB型血清培养液中比在添加FBS的培养液中增殖效果更加明显。Objective To observe the proliferation difference of human peripheral blood mononucelear cells ( PBMCs) which were stim-ulated with Mycobacterium tuberculosis ( M.tb) Whole Cell Lysate ( WCL) in human AB serum cell culture medium and fatal bovine serum ( FBS) cell culture medium.Methods PBMCs from healthy adults were stimulated by WCL or WCL with recombinant human interleukin 2 ( rhIL-2) or phytohemagglutinin ( PHA) or none as control in four groups.Each of the groups was cultured with RPMI-1640 medium containing 10%AB serum or 10%FBS for six days respectively.The proliferation of lymphocytes was carefully investiga-ted under the inverted microscope and flow cytometry.Results The proliferation of lymphocytes stimulated by WCL in medium with 10%AB serum is better than that in medium with 10%FBS (P〈0.05).Similar results were obtained when PBMCs were stimulated by WCL and rhIL-2 (P〈0.05) .Conclusion When human PBMCs were stimulated with M.tb WCL,the proliferation of lymphocytes was better in the culture medium supplemented with human AB serum than in that with FBS serum.
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