Baxevanis C N, Dedoussis G V, Papadopoulos N G, Missitzis I, Beroukas C, Stathopoulos G P, Papamichail M
Department of Immunology, Hellenic Anticancer Institute, Athens, Greece.
Cancer. 1995 Oct 1;76(7):1253-60. doi: 10.1002/1097-0142(19951001)76:7<1253::aid-cncr2820760724>3.0.co;2-f.
Lymphokine-activated killer (LAK) cell function can be generated in peripheral blood mononuclear cells (PBMC) after brief exposure of high dose interleukin-2 (IL-2) over the course of 1 or 2 days' culture in plain culture medium (IL-2-pulsed PBMC). The aim of the present study was to investigate the ability of granulocyte-macrophage-colony stimulating factor (GM-CSF) to augment LAK induction in low dose IL-2-pulsed PBMC derived from patients with cancer undergoing immunotherapy with IL-2.
Peripheral blood mononuclear cells were collected from patients with cancer receiving a 5-day cycle of local (intraperitoneal or intrapleural) infusions with IL-2. The cells were incubated with IL-2 in the presence or absence of GM-CSF for 1 hour and then tested as effectors against allogeneic tumor cells and LAK-sensitive cell lines.
Granulocyte-macrophage-colony stimulating factor at doses between 10 and 100 ng/ml was synergized with low dose IL-2 (100 IU/ml) in the generation of LAK activity in PBMC. Lymphokine-activated killer cell-mediated cytotoxicity derived from PBMC cultures incubated with IL-2 and GM-CSF was significantly higher (up to three-fold) compared with that generated with IL-2 alone. The GM-CSF-induced enhanced LAK activity was maintained when tested at day 5. GM-CSF increased the percentages of IL-2 receptor (R) positive (+) and CD8+ cells in the IL-2-pulsed PBMC. In contrast to CD56+ cells, highly purified CD8+ cells isolated from PBMC pulsed with IL-2 and GM-CSF responded with increased LAK activity, thus representing the cell-type that mediates the augmenting effect of GM-CSF. Major histocompatibility complex (MHC) molecules or the CD3 surface antigens were not involved in the GM-CSF-mediated enhancement of LAK induction because anti-MHC class I and class II monoclonal antibodies (MoAb) or MoAb against the CD3 molecules remained without any effect in this system. The GM-CSF-mediated LAK-enhancement was IL-2-dependent because MoAb against IL-2 receptor completely inhibited the generation of LAK activity.
The use of GM-CSF for the enhancement of IL-2-induced LAK activity in 1 hour cultures may improve clinical results in cancer immunotherapy. In addition, implementation of this procedure could eliminate the high cost of cell culture which usually accompanies IL-2/LAK cell therapy as well as eliminate the known toxic side effects associated with this kind of therapy.
在普通培养基中经过1或2天的培养，短暂暴露于高剂量白细胞介素-2（IL-2）后，外周血单个核细胞（PBMC）中可产生淋巴因子激活的杀伤（LAK）细胞功能（IL-2预刺激的PBMC）。本研究的目的是探讨粒细胞-巨噬细胞集落刺激因子（GM-CSF）增强低剂量IL-2预刺激的、来自接受IL-2免疫治疗的癌症患者的PBMC中LAK诱导的能力。
从接受为期5天的局部（腹腔内或胸腔内）IL-2输注周期的癌症患者中采集外周血单个核细胞。将细胞在有或无GM-CSF的情况下与IL-2孵育1小时，然后作为效应细胞针对同种异体肿瘤细胞和LAK敏感细胞系进行检测。
剂量在10至100 ng/ml之间的粒细胞-巨噬细胞集落刺激因子与低剂量IL-2（100 IU/ml）协同作用，在PBMC中产生LAK活性。与单独用IL-2培养产生的PBMC相比，用IL-2和GM-CSF培养产生的淋巴因子激活的杀伤细胞介导的细胞毒性显著更高（高达三倍）。在第5天进行检测时，GM-CSF诱导的增强的LAK活性得以维持。GM-CSF增加了IL-2预刺激的PBMC中IL-2受体（R）阳性（+）和CD8+细胞的百分比。与CD56+细胞不同，从用IL-2和GM-CSF预刺激的PBMC中分离出的高度纯化的CD8+细胞表现出增强的LAK活性，因此代表介导GM-CSF增强作用的细胞类型。主要组织相容性复合体（MHC）分子或CD3表面抗原不参与GM-CSF介导的LAK诱导增强，因为抗MHC I类和II类单克隆抗体（MoAb）或抗CD3分子的MoAb在该系统中没有任何作用。GM-CSF介导的LAK增强是IL-2依赖性的，因为抗IL-2受体的MoAb完全抑制了LAK活性的产生[14,15]。
在1小时培养中使用GM-CSF增强IL-2诱导的LAK活性可能会改善癌症免疫治疗的临床效果。此外，实施该程序可以消除通常伴随IL-2/LAK细胞疗法的细胞培养高成本，以及消除与这种疗法相关的已知毒副作用。
