Schuurman B, Heuff G, Beelen R H, Meyer S
Department of Surgical Oncology, Free University Hospital, Amsterdam, The Netherlands.
Cancer Immunol Immunother. 1994 Sep;39(3):179-84. doi: 10.1007/BF01533384.
In this study we investigated the effect of the cytokines human granulocyte/macrophage-colony-stimulating Factor (hGM-CSF) and interferon gamma (IFN gamma) on human Kupffer-cell-mediated cytotoxicity against the SW948 colon carcinoma cell line. Kupffer cells were isolated from small liver wedge biopsies, taken from 14 patients who had had abdominal surgery for colon carcinoma or partial hepatectomy. The cells were incubated with hGM-CSF (100 ng/ml), or with IFN gamma (100 U/ml) or with their combination and the percentage cytotoxicity was determined using a recently described modified assay. Additional experiments were performed with tumour-necrosis-factor-alpha (TNF alpha)-sensitive U937 cells as target. The TNF alpha secretion of Kupffer cells was measured and we evaluated the effect of TNF alpha on colon tumour targets. We performed human-Kupffer-cell-mediated cytotoxicity blocking experiments with anti-TNF alpha and used paraformaldehyde-fixed Kupffer cells to demonstrate lysis of TNF alpha-sensitive WEHI-164 cells and of SW948 cells. The overall cytotoxicity against SW948 caused by unactivated Kupffer cells (n = 14), and by Kupffer cells activated with hGM-CSF (n = 14), IFN gamma (n = 6) or their combination (n = 6) was respectively: 19.5 +/- 2.6%, 25.3 +/- 2.9%, 41 +/- 9.4% and 45.6 +/- 8% at E/T = 1 and 28.2 +/- 2.9%, 35.6 +/- 3.2%, 55.6 +/- 9.7% and 62.8% at E/T = 5. All differences were statistically significant (P < 0.05). No growth-promoting activity by hGM-CSF on the SW948 tumour cells was observed. U937 cells were highly susceptible to Kupffer-cell-mediated cytotoxicity. The TNF alpha secretion by human Kupffer cells increased in parallel to their cytotoxicity after incubation with these cytokines. Soluble TNF alpha had only a slight anti-proliferative effect on SW948 cells, while specific anti-TNF alpha blocked Kupffer cell cytotoxicity by up to 80%. Finally, paraformaldehyde-fixed Kupffer cells were able to lyse WEHI-164 and SW948 cells. This indicates that expression of cell-associated TNF alpha is the main cytolytic mechanism of human-Kupffer-cell-mediated cytotoxicity. The implications for the use of hGM-CSF and IFN gamma in vivo are discussed.
在本研究中，我们调查了细胞因子人粒细胞/巨噬细胞集落刺激因子（hGM-CSF）和干扰素γ（IFNγ）对人库普弗细胞介导的针对SW948结肠癌细胞系的细胞毒性作用。从14例因结肠癌接受腹部手术或部分肝切除术的患者所取的小块肝楔形活检组织中分离出库普弗细胞。将细胞与hGM-CSF（100 ng/ml）、或与IFNγ（100 U/ml）或二者联合孵育，并使用最近描述的改良检测方法测定细胞毒性百分比。另外以对肿瘤坏死因子α（TNFα）敏感的U937细胞作为靶细胞进行实验。测量库普弗细胞的TNFα分泌，并评估TNFα对结肠肿瘤靶细胞的作用。我们用抗TNFα进行人库普弗细胞介导的细胞毒性阻断实验，并使用多聚甲醛固定的库普弗细胞来证明对TNFα敏感的WEHI-164细胞和SW948细胞的裂解。未活化的库普弗细胞（n = 14）、用hGM-CSF活化的库普弗细胞（n = 14）、IFNγ活化的库普弗细胞（n = 6）或二者联合活化的库普弗细胞（n = 6）对SW948的总体细胞毒性在效靶比（E/T）为1时分别为：19.5±2.6%、25.3±2.9%、41±9.4%和45.6±8%，在E/T为5时分别为：28.2±2.9%、35.6±3.2%、55.6±9.7%和62.8%。所有差异均具有统计学意义（P < 0.05）。未观察到hGM-CSF对SW948肿瘤细胞有促生长活性。U937细胞对库普弗细胞介导的细胞毒性高度敏感。人库普弗细胞与这些细胞因子孵育后，其TNFα分泌与其细胞毒性呈平行增加。可溶性TNFα对SW948细胞仅有轻微的抗增殖作用，而特异性抗TNFα可使库普弗细胞细胞毒性阻断高达80%。最后，多聚甲醛固定的库普弗细胞能够裂解WEHI-164和SW948细胞。这表明细胞相关TNFα的表达是人库普弗细胞介导的细胞毒性的主要溶细胞机制。文中讨论了hGM-CSF和IFNγ在体内应用的意义。
