Kawai H, Komiyama A, Katoh M, Yabuhara A, Miyagawa Y, Akabane T
Department of Pediatrics, Shinshu University School of Medicine, Matsumoto, Japan.
Transfusion. 1988 Nov-Dec;28(6):531-5. doi: 10.1046/j.1537-2995.1988.28689059025.x.
Lymphokine-activated killer (LAK) and natural killer (NK) cells were studied for their capacity to retain cytotoxicity after cryopreservation. LAK cells were generated by a 4-day culture of lymphocytes with recombinant interleukin-2 (rIL-2). Cytotoxicity was measured by 51Cr-release assay at effector:target ratios of 10:1 to 80:1. Cryopreserved LAK cells retained 58.8 to 87.4 percent of cytotoxicity, as compared with that in fresh control cells. Cryopreserved NK cell activity against K562 and Molt-4 targets was 45.7 to 67.9 percent of the respective values of the fresh control cells. The responsiveness of NK cells to polyinosinic-polycytidilic acid (poly I:C), interferon-alpha (IFN-alpha), or rIL-2 remained intact. Activated NK cell activity after poly I:C or IFN-alpha stimulation and that after rIL-2 were, respectively, comparable to and higher than the endogenous NK cell activity of the fresh cells. The composition of lymphocyte subsets as determined by flow cytometry using monoclonal antibodies did not change after cryopreservation, indicating that cellular loss of the given subsets did not occur during the procedure. The retention of substantial levels of cytotoxicity in cryopreserved LAK and NK cells may make them promising candidates as cytotoxic effector cells.
研究了淋巴因子激活的杀伤细胞（LAK）和自然杀伤细胞（NK）在冷冻保存后保持细胞毒性的能力。LAK细胞通过淋巴细胞与重组白细胞介素-2（rIL-2）进行4天培养产生。通过51Cr释放试验在效应细胞：靶细胞比例为10:1至80:1的条件下测量细胞毒性。与新鲜对照细胞相比，冷冻保存的LAK细胞保留了58.8%至87.4%的细胞毒性。冷冻保存的NK细胞对K562和Molt-4靶标的活性分别为新鲜对照细胞相应值的45.7%至67.9%。NK细胞对聚肌苷酸-聚胞苷酸（poly I:C）、α干扰素（IFN-α）或rIL-2的反应性保持完整。poly I:C或IFN-α刺激后激活的NK细胞活性以及rIL-2刺激后激活的NK细胞活性分别与新鲜细胞的内源性NK细胞活性相当或高于其活性。使用单克隆抗体通过流式细胞术测定的淋巴细胞亚群组成在冷冻保存后没有变化，这表明在该过程中给定亚群的细胞没有损失。冷冻保存的LAK和NK细胞中大量细胞毒性的保留可能使它们成为有前景的细胞毒性效应细胞候选者。
