Ochoa A C, Gromo G, Alter B J, Sondel P M, Bach F H
J Immunol. 1987 Apr 15;138(8):2728-33.
Peripheral blood lymphocytes (PBL) cultured in interleukin 2 (IL 2)-containing medium in conventional tissue culture develop the ability to lyse fresh tumor cells; such cells are referred to as lymphokine-activated killer (LAK) cells. LAK activity peaks by day 5 of culture and declines rapidly thereafter. We studied culture conditions and signals that allow for long-term culture and expansion of cells with LAK activity. By culturing cells at relatively low densities and regularly replenishing medium and recombinant IL 2 (r-IL 2), LAK function is significantly higher as compared with short-term cultures, and remains present for at least 21 days while cell numbers undergo an average 100-fold expansion. By activating these cultures with anti-CD3 (OKT3) monoclonal antibody and r-IL 2, an approximately 1000-fold expansion in the cell number is obtained with maintenance of comparable levels of LAK activity. The exogenous addition of beta interleukin 1 (beta-IL 1), interferon-beta (IFN-beta) or interferon-gamma (IFN-gamma) can augment the lytic activity of cell populations expanded by anti-CD3 plus r-IL 2. These approaches may enable the in vitro generation from individual donors of much greater numbers of LAK cells for adoptive immunotherapy than can now be obtained with the 3 to 5 day in vitro culture systems.
在传统组织培养中，在含白细胞介素2（IL-2）的培养基中培养的外周血淋巴细胞（PBL）会产生裂解新鲜肿瘤细胞的能力；这类细胞被称为淋巴因子激活的杀伤细胞（LAK细胞）。LAK活性在培养第5天达到峰值，此后迅速下降。我们研究了能使具有LAK活性的细胞长期培养和扩增的培养条件及信号。通过以相对低密度培养细胞并定期补充培养基和重组IL-2（r-IL-2），与短期培养相比，LAK功能显著更高，并且在细胞数量平均扩增100倍的情况下，LAK功能至少维持21天。通过用抗CD3（OKT3）单克隆抗体和r-IL-2激活这些培养物，细胞数量可扩增约1000倍，同时维持相当水平的LAK活性。外源性添加β白细胞介素1（β-IL-1）、干扰素-β（IFN-β）或干扰素-γ（IFN-γ）可增强经抗CD3加r-IL-2扩增的细胞群体的裂解活性。这些方法可能使从个体供体体外产生比目前3至5天体外培养系统所能获得的数量多得多的LAK细胞用于过继性免疫治疗成为可能。
