Peptidoglycan-Like Components in Z-100, Extracted from Mycobacterium tuberculosis Strain Aoyama B, Increase IL-12p40 via NOD2

Abstract

Background. Z-100 is a hot-water extract of the human-type Mycobacterium tuberculosis strain Aoyama B. While Z-100’s macrophage-mediated immunomodulatory effects have been reported, the mechanistic details have not been fully clarified. Here, we studied the immunomodulatory effects of Z-100 on mouse bone marrow-derived cells, human CD14+ cells, and skin. Methods. Mouse bone marrow-derived cells and CD14+ cells were cultured in the presence of granulocyte-macrophage colony-stimulating factor, differentiated into macrophage-like cells, and then stimulated with Z-100. Furthermore, since Z-100 is subcutaneously administered clinically, we injected Z-100 into mice and measured gene expression in the skin. Results. While Z-100 stimulation increased the production of interleukin- (IL-) 12p40 and IL-1β in mouse bone marrow-derived macrophages, levels of IL-1β were low. In contrast, TNF-α production did not increase. Meanwhile, stimulation of human CD14+ cells with Z-100 increased production of IL-12p40, TNF-α, and IL-1β. Because Z-100 appeared to have the most stable effect on IL-12p40, we examined the components of Z-100 that induce IL-12p40 production. We found that Z-100 contained peptidoglycan-like components. In addition, an siRNA study showed that Z-100 increased the production of IL-12p40 via nucleotide-binding oligomerization domain 2 (NOD2). Further, subcutaneous administration of Z-100 to mice significantly elevated expression of IL-12p40 and IL-1β and showed a trend towards increasing TNF-α in the skin. Conclusion. Z-100 induced the production of immunomodulatory cytokines from various types of macrophages and specifically increased IL-12p40 production through peptidoglycan-like components via NOD2.

Potential of Z-100, extracted from Mycobacterium tuberculosis strain Aoyama B, as a hot tumor inducer

“…We previously reported that Z-100 increases IL-12p40 production [13]. Therefore, we examined whether the antitumor effects of Z-100 were abolished in IL-12p40 KO mice and found that, indeed, the antitumor effects of Z-100 were absent in these animals, as shown in Fig.…”

“…Immunotherapies that include an anti-PD-1 antibody have been reported to be highly effective against hot tumors [1,4,21], which may explain why the combination of Z-100 and anti-PD-1 antibody was effective in the report by Kobayashi et al [14]. Further, we previously reported that Z-100 increases IL-12p40 production [13]. IL-12p40 is known bind to IL-12p35 to form the heterodimer IL-12, which is involved in antitumor immunity [22].…”

“…Given that Z-100 has very mild toxicity in clinical practice [17], its effects may differ from those of IL-12 administration. Z-100 stimulation has been reported to increase TNF-α, IL-1β, IFN-γ, and IL-2 from immune cells [9,10,13,31], the production of which may underlie the different activities of Z-100 and IL-12. We also previously reported that Z-100 increases IL-12p40 production from M1-like macrophages differentiated from mouse bone marrow cells and human CD14 + cells in the presence of GM-CSF [13].…”

“…Several reports have also described its antitumor effects [7][8][9][10][11]. According to a handful of studies on the mechanism of Z-100, it exerts its effects by inducing cytokines [10,12,13]. In particular, Z-100-mediated production of IL-12p40 from macrophages is thought to contribute to the drug's anti-cancer effects [10,12,13].…”

“…According to a handful of studies on the mechanism of Z-100, it exerts its effects by inducing cytokines [10,12,13]. In particular, Z-100-mediated production of IL-12p40 from macrophages is thought to contribute to the drug's anti-cancer effects [10,12,13]. A recent clinical study by Kobayashi et al reported that combined use of Z-100 with an anti-PD-1 antibody was effective for treating lung cancer [14].…”