从分枝杆菌菌株 Aoyama B 中提取的 Z-100 通过核苷酸结合寡聚化结构域 2（Nod2）依赖性 NF-κB 激活促进 RAW264.7 细胞中 TNF-α 的产生。
Z-100, extracted from Mycobacterium tuberculosis strain Aoyama B, promotes TNF-α production via nucleotide-binding oligomerization domain containing 2 (Nod2)-dependent NF-κB activation in RAW264.7 cells.
机构信息
Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
出版信息
Mol Immunol. 2015 Mar;64(1):218-27. doi: 10.1016/j.molimm.2014.11.017. Epub 2014 Dec 8.
Macrophages are a major component of the innate immune system, and the cytokines they secrete are involved in antitumor responses. Z-100 is obtained from hot-water extract of human-type Mycobacterium tuberculosis strain Aoyama B and activates the innate immune response. However, while Z-100 is known to modulate macrophage activity, the mechanism behind this modulation is not fully understood. We evaluated the effects of Z-100 on the murine macrophage cell line RAW264.7. Tumor necrosis factor-alpha (TNF-α) production from RAW264.7 cells was strongly induced by Z-100 and interferon-gamma (IFN-γ) stimulation but only weakly induced by Z-100 alone. Quantitative gene expression analysis showed that nucleotide-binding oligomerization domain containing 2 (Nod2) expression was up-regulated by IFN-γ treatment in RAW264.7 cells while Z-100-induced TNF-α production was attenuated by Nod2 gene silencing. Further, componential analysis demonstrated that muramic acid and amino acids distinctive of muramyl dipeptide (MDP) were contained within Z-100 and Z-100Fr I, the low-molecular-weight fraction containing components <3 kDa in size. In addition, Z-100Fr I enhanced TNF-α production in RAW264.7 cells and promoted NOD2-dependent nuclear factor-kappa B (NF-κB) activation in murine NOD2-expressing SEAP reporter HEK293 (HEK-Blue-mNOD2) cells. Taken together, these results suggest that Z-100 contains MDP-like molecules and augments NF-κB signaling via the direct activation of Nod2 in macrophages, which might be one mechanism driving the innate immune responses induced by Z-100 in cancer immunotherapy.
巨噬细胞是先天免疫系统的主要组成部分，它们分泌的细胞因子参与抗肿瘤反应。Z-100 是人型结核分枝杆菌 Aoyama B 热水提取物的产物，可激活先天免疫反应。然而，虽然已知 Z-100 可调节巨噬细胞活性，但这种调节的机制尚不完全清楚。我们评估了 Z-100 对鼠源巨噬细胞系 RAW264.7 的影响。Z-100 和干扰素-γ（IFN-γ）刺激强烈诱导 RAW264.7 细胞产生肿瘤坏死因子-α（TNF-α），而单独 Z-100 诱导作用较弱。定量基因表达分析显示，IFN-γ 处理可上调 RAW264.7 细胞中核苷酸结合寡聚化结构域包含蛋白 2（Nod2）的表达，而 Z-100 诱导的 TNF-α 产生可被 Nod2 基因沉默减弱。进一步的成分分析表明，Z-100 和低分子量部分 Z-100Fr I 中含有 N-乙酰胞壁酸和独特的 N-乙酰氨基葡萄糖，这些物质是 N-乙酰胞壁酰二肽（MDP）的组成部分。此外，Z-100Fr I 增强 RAW264.7 细胞中 TNF-α 的产生，并促进表达 NOD2 的 SEAP 报告基因 HEK293（HEK-Blue-mNOD2）细胞中 NOD2 依赖性核因子-κB（NF-κB）的激活。综上所述，这些结果表明 Z-100 含有 MDP 样分子，并通过直接激活巨噬细胞中的 Nod2 增强 NF-κB 信号通路，这可能是 Z-100 在癌症免疫治疗中诱导先天免疫反应的一种机制。
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