TLR4 and NLRP3 inflammasome activation in monocytes by N-propionyl cysteaminylphenol-maleimide-dextran (NPCMD)

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• 作者：Yu Mizote ; Kazumasa Wakamatsu ; Shosuke Ito ; Akiko Uenaka ; Yoshihiro Ohue ; Koji Kurose ; Midori Isobe ; Akira Ito ; Yasuaki Tamura ; Hiroyuki Honda ; Toshiharu Yamashita ; Satoshi Nohara ; Mikio Oka ; Kowichi Jimbow ; Eiichi Nakayama
• 关键词：4-S-CAP ; 4-S-cysteaminylphenol ; alum ; aluminum hydroxide ; APDC ; (2R ; 4R)-4-aminopyrrolidine-2 ; 4-dicarboxylic acid ; ATP ; adenosine triphosphate ; CBA ; cytometric bead array ; CMD ; carboxymethyl dextran ; DAMP ; danger-associated molecular pattern ; defASC ; ASC-deficient THP-1 ; defNLRP3 ; NLRP3-deficient THP-1 ; FBS ; fetal bovine serum ; LAL ; limulus amebocyte lysate ; LPS ; lipopolysaccharide ; MIL ; maleimide linker ; MIL-CMD ; maleimide linker conjugated carboxymethyl dextran ; MTT ; methylthiazole tetrazolium
• 刊名：Journal of Dermatological Science
• 出版年：March, 2014
• 年：2014
• 卷：73
• 期：3
• 页码：209-215
• 全文大小：1193 K

文摘

Background

N-propionyl cysteaminylphenol-maleimide-dextran (NPCMD) is a toxic tyrosinase substrate developed to treat melanoma.

Objective

We investigated the effect of NPCMD on innate immune responses in monocytes.

Methods

CD14⁺ monocytes and a monocytic cell line, THP-1, were stimulated with NPCMD in vitro. Cytokines in the culture supernatants were determined by ELISA and flow cytometry.

Results

NPCMD stimulated CD14⁺ monocytes and THP-1 cells to secrete TNF伪, IL-6 and IL-8, but not IL-10 or IL-12. TNF伪 secretion from THP-1 cells stimulated with NPCMD was inhibited by addition of an anti-TLR4 mAb in culture. Moreover, NPCMD stimulated production of pro-IL-1尾 in CD14⁺ monocytes and monocytic cell line THP-1 cells and activated the NLRP3-inflammasome, resulting in production of mature IL-1尾. Use of ASC and NLRP3-deficient THP-1 cell lines established involvement of the NLRP3 inflammasome in an IL-1尾 secretion in treatment with NPCMD. Inhibition of IL-1尾 secretion by an endocytosis inhibitor, cytochalasin B, and a lysosomal enzyme cathepsin B inhibitor, CA-074 Me, suggested the involvement of lysosomal rupture and leakage of cathepsin B into the cytosol in NLRP3 activation by NPCMD.

Conclusion

The immunopotentiating effect of NPCMD mediated by TLR4 and NLRP3 inflammasome activation could be useful for eliciting effective adaptive immune responses against melanoma and other tumors.