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本文将新分离的成人外周血单个核细胞(PBMC)分3组培养:(1)用抗CD3单抗(αCD3)、rIL-2共同刺激PBMC,诱生、扩增CD3AK;(2)用PHA预刺激PBMC48小时,再用含rIL-2的全培养基培养,诱生、扩增PHA-LAK细胞;(3)用PHA预刺激PBMC48小时,再用αCD3、rIL-2协同刺激,诱生、扩增新型效应细胞PHA-αCD3LAK.利用活细胞计数、MTT、ELISA、A-PAAP等方法对3组效应细胞的增殖动力学、细胞毒活性、细胞表型、mIL-2Rα的表达及培养上清液中sIL-2R和IL-2的含量作了测定和比较.结果表明:PHA-αCD3LAK、PHA-LAK和CD3AK 3组细胞(1)均为异质性细胞群体,PHA-αCD3LAK以CD3~+、CD8~+T细胞为主,含有较多的CTL细胞,(2)其增殖曲线及mIL-2Rα的表达均于培养的第6天达到高峰,其峰值和扩增倍数均为PHA-αCD3LAK>PHA-LAK>CD3AK(P<0.05),(3)均可介导MHC非限制的细胞毒性,对K562细胞和
In this article, newly isolated human peripheral blood mononuclear cells (PBMCs) were cultured in 3 groups: (1) PBMCs were stimulated with anti-CD3 monoclonal antibody (αCD3) and rIL-2 to induce and amplify CD3AK; (2) PHA was used; PBMCs were pre-stimulated for 48 hours and then cultured with whole medium containing rIL-2 to induce and amplify PHA-LAK cells; (3) PBMCs were pre-stimulated with PHA for 48 hours, followed by synergistic stimulation with αCD3 and rIL-2. To amplify novel effector cells PHA-αCD3LAK. The proliferation kinetics, cytotoxicity, cell phenotype, expression of mIL-2Rα and culture supernatant of the three groups of effector cells were measured by viable cell count, MTT, ELISA, and A-PAAP. The contents of sIL-2R and IL-2 in the fluid were measured and compared. The results showed that: PHA-αCD3LAK, PHA-LAK and CD3AK 3 groups of cells (1) are all heterogeneous cell populations, PHA-αCD3LAK with CD3~+ , CD8~+ T cells were predominant, containing more CTL cells, (2) the proliferation curve and the expression of mIL-2Rα all peaked on the 6th day of culture, and the peak and amplification multiples were both PHA-αCD3LAK> PHA-LAK>CD3AK (P<0.05), (3) can all mediate the unrestricted cytotoxicity of MHC on K562 cells and