Lafreniere R, Borkenhagen K, Bryant L D, Huchcroft S
Division of Surgical Oncology, University of Calgary, Alberta, Canada.
Cancer Res. 1989 May 1;49(9):2409-14.
It has been shown that the systemic administration of lymphokine-activated killer (LAK) cells with recombinant interleukin 2 (RIL-2) is effective in reducing the number of established pulmonary and hepatic metastases in murine models. Similarly, this modality of therapy has been proven effective against certain selected human tumors as well. In view of the rising concern with transmission of virally related communicable diseases such as hepatitis and AIDS, we have undertaken the evaluation of a serum-free medium (AIM V) for the generation and expansion of murine LAK cells for use in in vivo tumor immunotherapy against murine hepatic metastases. Day 3 LAK cells generated in AIM V medium demonstrated a greater percentage of viable cells than cells generated in serum containing complete medium (CM) (mean percentage of yield, 59 versus 25%, AIM V medium versus CM, respectively, P less than 0.001, N = 6 consecutive experiments). When day 3 LAK cells were transferred to new medium (CM to CM and AIM V to AIM V), a highly reproducible expansion of these cells was demonstrated which was significantly better for cells expanded in AIM V medium versus cells expanded in CM (mean fold expansion on day 21 of culture; 201 versus 54, AIM V medium versus CM, respectively, P less than 0.005, N = 4 consecutive experiments). When day 3 LAK cells, day 5 expanded LAK cells, and day 13 expanded LAK cells grown in CM or in AIM V medium were given in vivo with RIL-2 to mice harboring hepatic metastases, cells grown in AIM V medium demonstrated an increased antitumor activity compared to cells grown in CM. As an example in experiment 1, the mean number of metastases with day 5 expanded LAK cells grown in CM and given with RIL-2 was 47 while the mean number of metastases with day 5 expanded LAK cells grown in AIM V medium and given with RIL-2 was 5 (P less than 0.002). These experiments demonstrate that AIM V medium can be utilized to generate greater numbers of murine LAK cells with enhanced in vivo antitumor activity compared to cells generated in CM. These findings could be applied to the expansion of cytotoxic cells for human antitumor therapy.
已表明，在小鼠模型中，全身性给予淋巴因子激活的杀伤细胞（LAK）和重组白细胞介素2（RIL-2）可有效减少已形成的肺和肝转移灶数量。同样，这种治疗方式也已被证明对某些特定的人类肿瘤有效。鉴于人们对诸如肝炎和艾滋病等病毒相关传染病传播的担忧日益增加，我们对一种无血清培养基（AIM V）进行了评估，该培养基用于生成和扩增小鼠LAK细胞，以用于针对小鼠肝转移灶的体内肿瘤免疫治疗。在AIM V培养基中生成的第3天LAK细胞显示出比在含血清的完全培养基（CM）中生成的细胞更高的活细胞百分比（平均产率百分比分别为59%对25%，AIM V培养基对CM，P小于0.001，N = 6次连续实验）。当第3天的LAK细胞转移到新培养基中（CM到CM以及AIM V到AIM V）时，这些细胞显示出高度可重复的扩增，在AIM V培养基中扩增的细胞比在CM中扩增的细胞明显更好（培养第21天的平均扩增倍数；分别为201对54，AIM V培养基对CM，P小于0.005，N = 4次连续实验）。当将在CM或AIM V培养基中生长的第3天LAK细胞、第5天扩增的LAK细胞和第13天扩增的LAK细胞与RIL-2一起在体内给予患有肝转移的小鼠时，在AIM V培养基中生长的细胞与在CM中生长的细胞相比显示出增强的抗肿瘤活性。例如在实验1中，在CM中生长并与RIL-2一起给予的第5天扩增的LAK细胞的转移灶平均数为47，而在AIM V培养基中生长并与RIL-2一起给予的第5天扩增的LAK细胞的转移灶平均数为5（P小于0.002）。这些实验表明，与在CM中生成的细胞相比，AIM V培养基可用于生成更多具有增强的体内抗肿瘤活性的小鼠LAK细胞。这些发现可应用于扩增用于人类抗肿瘤治疗的细胞毒性细胞。
