Falk L A, Hogan M M, Vogel S N
Department of Microbiology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814.
J Leukoc Biol. 1988 May;43(5):471-6. doi: 10.1002/jlb.43.5.471.
Bone marrow-derived cells from C3H/HeJ mice were cultured in the presence of recombinant murine granulocyte-macrophage colony-stimulating factor (rGM-CSF) or highly purified murine macrophage colony-stimulating factor (CSF-1) for 7 days. Following this 7-day culture period, mature macrophages were harvested and replated at precise densities in the absence of exogenous rGM-CSF or CSF-1, and assayed in a two-signal tumoricidal assay. Cultures were stimulated with medium only or with combinations of recombinant interferon-gamma (rIFN-gamma) as the "priming" signal, and/or butanol-extracted lipopolysaccharide (But-LPS) as the "triggering" signal for 24 hr. At this time, 51Cr-labeled, P815 tumor target cells were added, and the percent tumor cell cytotoxicity was determined after 16 hr. Macrophages derived under the influence of rGM-CSF exhibited significant tumoricidal capacity with medium alone (16 +/- 5%). The addition of "priming" signal only (i.e., rIFN-gamma, 10.0 U/ml) significantly increased tumoricidal capacity to 31 +/- 9%. Treatment with But-LPS alone did not alter the basal tumoricidal activity of rGM-CSF-derived macrophages. Combinations of rIFN-gamma (10.0 U/ml) and But-LPS (0.5-5.0 micrograms/ml) generated highly tumoricidal macrophages (50-60% tumor cell cytotoxicity). In contrast, medium-treated CSF-1-derived macrophages exhibited a significantly lower basal level of tumor cytotoxicity (6 +/- 3%). Unlike rGM-CSF-derived macrophages, treatment of CSF-1-derived macrophages with high concentrations of rIFN-gamma alone did not increase significantly the level of cytotoxicity above that of medium-treated cultures. However, CSF-1-derived macrophages responded to the highest concentrations of But-LPS (5.0 micrograms/ml) to increase tumoricidal activity from 6 +/- 3% to 17 +/- 5%. Optimal tumoricidal activity (44 +/- 17%) was observed when CSF-1-derived macrophages were treated simultaneously with high concentrations of both rIFN-gamma and But-LPS. Thus, macrophages derived from bone marrow progenitors in either rGM-CSF or CSF-1 exhibited tumoricidal capacities that differed in basal activity as well as in their requirements for and sensitivities to "priming" and "triggering" signals.
将C3H/HeJ小鼠的骨髓来源细胞在重组小鼠粒细胞 - 巨噬细胞集落刺激因子（rGM - CSF）或高度纯化的小鼠巨噬细胞集落刺激因子（CSF - 1）存在的情况下培养7天。在这7天的培养期后，收获成熟巨噬细胞，并在不存在外源性rGM - CSF或CSF - 1的情况下以精确密度重新接种，并在双信号杀瘤试验中进行检测。培养物仅用培养基刺激，或用重组干扰素 - γ（rIFN - γ）作为“启动”信号和/或丁醇提取的脂多糖（But - LPS）作为“触发”信号的组合刺激24小时。此时，加入51Cr标记的P815肿瘤靶细胞，并在16小时后测定肿瘤细胞细胞毒性百分比。在rGM - CSF影响下产生的巨噬细胞单独用培养基时表现出显著的杀瘤能力（16±5％）。仅添加“启动”信号（即rIFN - γ，10.0 U/ml）可使杀瘤能力显著提高至31±9％。单独用But - LPS处理不会改变rGM - CSF来源巨噬细胞的基础杀瘤活性。rIFN - γ（10.0 U/ml）和But - LPS（0.5 - 5.0μg/ml）的组合产生高杀瘤活性的巨噬细胞（50 - 60％肿瘤细胞细胞毒性）。相比之下，用培养基处理的CSF - 1来源巨噬细胞表现出显著较低的基础肿瘤细胞毒性水平（6±3％）。与rGM - CSF来源的巨噬细胞不同，单独用高浓度rIFN - γ处理CSF - 1来源的巨噬细胞不会使细胞毒性水平比用培养基处理的培养物显著增加。然而，CSF - 1来源的巨噬细胞对最高浓度的But - LPS（5.0μg/ml）有反应，使杀瘤活性从6±3％增加到17±5％。当CSF - 1来源的巨噬细胞同时用高浓度的rIFN - γ和But - LPS处理时，观察到最佳杀瘤活性（44±17％）。因此，源自rGM - CSF或CSF - 1中骨髓祖细胞的巨噬细胞在基础活性以及对“启动”和“触发”信号的需求和敏感性方面表现出不同的杀瘤能力。
