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

• Published in: Molecular immunology 2015-03, Vol.64 (1), p.218-227
• Main Authors: Katsunuma, Kokichi, Yoshinaga, Koji, Ohira, Yuta, Eta, Runa, Sato, Takanori, Horii, Takayuki, Tanaka, Takao, Takei, Mineo, Seto, Koichi
• Format: Article
• Language: English
• Subjects: Acetylmuramyl-Alanyl-Isoglutamine - pharmacology; Animals; Cell Line; Lipids - chemistry; Lipids - isolation & purification; Lipids - pharmacology; Macrophages; Macrophages - drug effects; Macrophages - metabolism; Mannans - chemistry; Mannans - isolation & purification; Mannans - pharmacology; Mice; Molecular Weight; Mycobacterium tuberculosis; Mycobacterium tuberculosis - chemistry; NF-kappa B - metabolism; NF-κB; Nod2; Nod2 Signaling Adaptor Protein - metabolism; Protein Transport - drug effects; Signal Transduction - drug effects; Tumor Necrosis Factor-alpha - biosynthesis; Z-100
• ISSN: 0161-5890; 1872-9142
• DOI: 10.1016/j.molimm.2014.11.017

•TNF-α production from RAW264.7 cells was induced by Z-100 and IFN-γ stimulation.•Nod2 expression was up-regulated by IFN-γ treatment in RAW264.7 cells.•Z-100-induced TNF-α production was attenuated by Nod2 gene silencing.•Z-100 activates Nod2-dependent NF-κB signaling. 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