Itoh K, Pellis N R, Balch C M
Division of Surgery, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Cancer Immunol Immunother. 1989;29(1):57-62. doi: 10.1007/BF00199917.
Both phytohemagglutinin-induced cytotoxicity and recombinant-interleukin-2 (rIL-2)-induced lymphokine-activated killer (LAK) activity against noncultured melanoma cells were significantly reduced when peripheral blood mononuclear cells (PBMC) from patients with metastatic melanoma were incubated in RPMI medium 1640 and 10% autologous human serum instead of 10% fetal calf serum, while serum from either healthy donors or patients with primary melanoma did not affect the level of cytotoxicity. The serum-mediated suppression was not restricted by major histocompatibility complex and was time-dependent. Addition of 10% human serum from the patients with metastatic melanoma [HS-Pt(m)] to the culture of PBMC with rIL-2 at the same time or 1 day after incubation significantly inhibited LAK activity. However, addition of 10% HS-Pt(m) 2 or 3 days after incubation did not inhibit LAK activity. Incubation of PBMC for 2 h with a high dose (10(4) U/ml) of rIL-2 in the presence of 10% HS-Pt(m), followed by incubation in the absence of either rIL-2 or HS-Pt(m), did not affect LAK cell activity. These results suggest that HS-Pt(m) inhibits the early stage of LAK cell differentiation, rather than the binding of rIL-2 to PBMC or a later stage in the differentiation. In contrast to PBMC, monocyte-depleted peripheral blood lymphocytes exhibited comparable levels of LAK activity when cultured with rIL-2 either in 10% fetal calf serum, 10% human serum from healthy donors or 10% HS-Pt(m). Addition of purified autologous monocytes to the culture of monocyte-depleted peripheral blood lymphocytes with rIL-2 suppressed LAK cell induction when 10% HS-Pt(m) was present. Thus serum-mediated suppression of LAK cell induction is largely dependent on the presence of monocytes, which may produce a secondary inhibitor that acts on lymphocytes. Addition of indomethacin to the culture did not reverse this monocyte-dependent serum-mediated suppression in a majority of cases, suggesting that prostaglandin E2 does not have a major role in the suppression.
当将转移性黑色素瘤患者的外周血单个核细胞（PBMC）置于RPMI 1640培养基及10%自体人血清而非10%胎牛血清中培养时，植物血凝素诱导的细胞毒性以及重组白细胞介素-2（rIL-2）诱导的针对未培养黑色素瘤细胞的淋巴因子激活的杀伤细胞（LAK）活性均显著降低，而来自健康供体或原发性黑色素瘤患者的血清则不影响细胞毒性水平。血清介导的抑制不受主要组织相容性复合体限制且具有时间依赖性。在培养PBMC并加入rIL-2的同时或培养1天后加入10%转移性黑色素瘤患者的人血清[HS-Pt(m)]，可显著抑制LAK活性。然而，在培养2天或3天后加入10% HS-Pt(m)则不抑制LAK活性。在10% HS-Pt(m)存在的情况下，将PBMC与高剂量（10⁴ U/ml）的rIL-2孵育2小时，随后在无rIL-2或HS-Pt(m)的情况下孵育，并不影响LAK细胞活性。这些结果表明，HS-Pt(m)抑制LAK细胞分化的早期阶段，而非rIL-2与PBMC的结合或分化的后期阶段。与PBMC不同，去除单核细胞的外周血淋巴细胞在10%胎牛血清、10%健康供体人血清或10% HS-Pt(m)中与rIL-2一起培养时，表现出相当水平的LAK活性。当存在10% HS-Pt(m)时，向去除单核细胞的外周血淋巴细胞与rIL-2的培养物中加入纯化的自体单核细胞会抑制LAK细胞的诱导。因此，血清介导的LAK细胞诱导抑制在很大程度上取决于单核细胞的存在，单核细胞可能产生一种作用于淋巴细胞的二级抑制剂。在大多数情况下，向培养物中加入吲哚美辛并不能逆转这种单核细胞依赖性血清介导的抑制，这表明前列腺素E2在抑制中不起主要作用。
