Ströhlein Michael Alfred, Grützner Klaus-Uwe, Schildberg Friedrich-Wilhelm, Heiss Markus Maria
Department of Surgery, Klinikum Grosshadern, Ludwig-Maximilians-University, Marchioninstrasse 15, 81377 Munich, Germany.
Cancer Immunol Immunother. 2002 Nov;51(9):505-12. doi: 10.1007/s00262-002-0310-6. Epub 2002 Sep 4.
The aim of this study was to investigate the cell-mediated immune response in 14 patients undergoing curative resection for a gastrointestinal tumor by the induction of peripheral blood mononuclear cell (PBMC)-mediated immune activity against autologous tumour cells. PBMC were stimulated by interleukin-12 (IL-12; 100 IU/ml) and IL-2 (1,000 IU/ml) without contact with tumour cells for 36 h. Specific cytotoxic activity against autologous tumour cells (auTu), natural killer (NK)-sensitive cells (K562) and allogeneic tumour cells (RF48/HT29) was determined by fluorescence cytotoxicity assay. Additionally, inhibition experiments using the mononuclear antibodies (mAb) FMC16 and W6/32 against major histocompatibility complex I (MHC I) on autologous tumour cells were performed in order to determine the involvement of specific T lymphocytes. The cytotoxic activity of unstimulated PBMC did not differ between the three target cells. IL-12 caused a 3.2-fold increase in activity against auTu ( P=0.002). In contrast, after stimulation with IL-2, only a slight increase in activity was observed. After IL-12 stimulation, cytotoxic activity against auTu was 2.5- to 2.7-fold higher than the corresponding activity against K562/allogeneic tumour cells ( P=0.002/ P=0.006). After blocking of the MHC I complex on auTu by FMC 16 or W6/32 mAb, a 62.9%/74.4% reduction in the specific cytotoxicity of IL-12-stimulated PBMC was found. In summary, IL-12 induced an effective immune response against auTu, which was partly mediated by specific cytotoxic T lymphocytes (CTL). It was considered that de novo generation of this activity during 36 h incubation without antigen contact was hardly possible, but that the observed induction of effective anti-tumor cytotoxicity was rather based on the re-activation of a pre-existing immune potential from the tumour-host interaction. These findings indicate the existence of an autologous anti-tumor immune response following curative resection in patients undergoing surgery for solid tumours, which might influence the development of tumour recurrence from disseminated tumour cells. Making use of this capacity could constitute an attractive immunotherapeutical approach for curatively operated tumour patients.
本研究的目的是通过诱导外周血单个核细胞（PBMC）介导的针对自体肿瘤细胞的免疫活性，来调查14例接受胃肠道肿瘤根治性切除术患者的细胞介导免疫反应。PBMC在不接触肿瘤细胞的情况下，用白细胞介素-12（IL-12；100 IU/ml）和IL-2（1000 IU/ml）刺激36小时。通过荧光细胞毒性测定法测定针对自体肿瘤细胞（auTu）、自然杀伤（NK）敏感细胞（K562）和同种异体肿瘤细胞（RF48/HT29）的特异性细胞毒性活性。此外，为了确定特异性T淋巴细胞的参与情况，对自体肿瘤细胞进行了使用针对主要组织相容性复合体I（MHC I）的单克隆抗体（mAb）FMC16和W6/32的抑制实验。未刺激的PBMC在三种靶细胞之间的细胞毒性活性没有差异。IL-12使针对auTu的活性增加了3.2倍（P = 0.002）。相比之下，用IL-2刺激后，仅观察到活性略有增加。IL-12刺激后，针对auTu的细胞毒性活性比针对K562/同种异体肿瘤细胞的相应活性高2.5至2.7倍（P = 0.002/P = 0.006）。在用FMC 16或W6/32 mAb阻断auTu上的MHC I复合体后，发现IL-12刺激的PBMC的特异性细胞毒性降低了62.9%/74.4%。总之，IL-12诱导了针对auTu的有效免疫反应，这部分是由特异性细胞毒性T淋巴细胞（CTL）介导的。据认为，在没有抗原接触的36小时孵育期间从头产生这种活性几乎是不可能的，但观察到的有效抗肿瘤细胞毒性的诱导相当程度上是基于肿瘤-宿主相互作用中预先存在的免疫潜能的重新激活。这些发现表明，接受实体肿瘤手术的患者在根治性切除后存在自体抗肿瘤免疫反应，这可能会影响播散性肿瘤细胞导致的肿瘤复发的发展。利用这种能力可能构成一种有吸引力的针对接受根治性手术的肿瘤患者的免疫治疗方法。
