Yang S C, Fry K D, Grimm E A, Roth J A
Department of Thoracic Surgery, University of Texas M.D. Anderson Cancer Center, Houston 77030.
J Immunother (1991). 1991 Oct;10(5):326-35. doi: 10.1097/00002371-199110000-00004.
We studied the phenotype and antitumor cytolytic activity of splenocytes from mice with K-1735 pulmonary metastases and tumor-infiltrating lymphocytes (TILs) from these metastases following treatment with anti-CD3, IL-2, and TNF combination immunotherapy. Mice were injected with 5 x 10(4) tumor cells and received a single 5 micrograms i.p. dose of anti-CD3 on day 3, followed by either IL-2 alone or fourfold less IL-2 with TNF (25,000 U/day) given at 3-day intervals. A single dose of anti-CD3 followed by low-dose IL-2 and TNF caused the greatest reduction in metastases compared to anti-CD3 alone, higher doses of IL-2 alone, or IL-2 + TNF. Reduction in metastases (greater than 80%) using the three agents was equal to or exceeded that achieved by ninefold higher concentrations of IL-2 alone. Treatment with anti-CD3 + IL-2 and TNF significantly prolonged survival, and resulted in 60% of mice achieving long-term survival greater than 120 days. This was superior to single agents or other combinations with the three agents causing a synergistic rather than additive effect. The anti-CD3-activated splenocytes were a heterogeneous population of T cells, with an increased number of CD8+ cells compared to splenocytes from mice treated with high doses of IL-2 alone. Analysis of TILs showed a greater proportion of CD8+ cells in anti-CD3-treated mice compared to IL-2 alone, but a lower proportion of CD4+ cells. Lymphokine-activated killer (LAK) and natural killer (NK) activities of both splenocytes and TILs in vitro increased following anti-CD3/IL-2 + TNF treatment, and were consistently greater than that generated with four times more IL-2 alone. TNF appeared to potentiate cytolytic activity rather than affect phenotypic changes. These results indicate that the sequential use of anti-CD3, IL-2, and TNF for LAK induction and maintenance potentiates antitumor activity, and suggests novel strategies for combination immunotherapy.
我们研究了K - 1735肺转移小鼠脾细胞的表型和抗肿瘤细胞溶解活性,以及这些转移灶的肿瘤浸润淋巴细胞(TILs)在接受抗CD3、白细胞介素-2(IL - 2)和肿瘤坏死因子(TNF)联合免疫治疗后的情况。给小鼠注射5×10⁴个肿瘤细胞,并在第3天腹腔注射5微克单剂量的抗CD3,随后要么单独给予IL - 2,要么给予剂量为单独使用IL - 2时四倍少的IL - 2并每隔3天给予TNF(25,000单位/天)。与单独使用抗CD3、更高剂量的单独IL - 2或IL - 2 + TNF相比,单剂量抗CD3后给予低剂量IL - 2和TNF导致转移灶减少最多。使用这三种药物使转移灶减少(大于80%)等同于或超过单独使用九倍高浓度IL - 2所达到的效果。抗CD3 + IL - 2和TNF治疗显著延长了生存期,60%的小鼠实现了大于120天的长期存活。这优于单一药物或这三种药物的其他组合,产生的是协同而非相加效应。抗CD3激活的脾细胞是异质性的T细胞群体,与单独用高剂量IL - 2治疗的小鼠的脾细胞相比,CD8⁺细胞数量增加。对TILs的分析表明,与单独使用IL - 2相比,抗CD3治疗的小鼠中CD8⁺细胞比例更高,但CD4⁺细胞比例更低。体外抗CD3/IL - 2 + TNF治疗后,脾细胞和TILs的淋巴因子激活的杀伤细胞(LAK)和自然杀伤细胞(NK)活性均增加,且始终大于单独使用四倍剂量IL - 2所产生的活性。TNF似乎增强了细胞溶解活性而非影响表型变化。这些结果表明,序贯使用抗CD3、IL - 2和TNF进行LAK诱导和维持可增强抗肿瘤活性,并提示了联合免疫治疗的新策略。