title: 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.
abstract: 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.
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan. Electronic address: koukichi-katsunuma@zeria.co.jp.
authors.forename: Kokichi
authors.initials: K
authors.lastname: Katsunuma
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Yoshinaga
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Ohira
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
authors.forename: Runa
authors.initials: R
authors.lastname: Eta
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Sato
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Horii
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
authors.forename: Mineo
authors.initials: M
authors.lastname: Takei
authors.affiliation: Central Research Laboratories, Zeria Pharmaceutical Co., Ltd., 2512-1, Numagami, Oshikiri, Kumagaya City 360-0111, Saitama, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Seto
mesh_terms: D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D008055:Lipids / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D008351:Mannans / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D051379:Mice; D008970:Molecular Weight; D009169:Mycobacterium tuberculosis / Q000737:chemistry*; D016328:NF-kappa B / Q000378:metabolism*; D053473:Nod2 Signaling Adaptor Protein / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
chemical_list: D008055:Lipids; D008351:Mannans; D016328:NF-kappa B; D053473:Nod2 Signaling Adaptor Protein; D014409:Tumor Necrosis Factor-alpha; C031208:specific substance maruyama; D000119:Acetylmuramyl-Alanyl-Isoglutamine
keywords: Macrophages; NF-κB; Nod2; Z-100