The Chemotherapeutic Agent DMXAA as a Unique IRF3-Dependent Type-2 Vaccine Adjuvant

Choon Kit Tang,Keiichi Ohata,Junichi Ito,Etsushi Kuroda,Benoit H Dessailly,Cevayir Coban,Ken J Ishii,Nao Jounai,Taiki Aoshi,Shizuo Akira,Kouji Kobiyama,Kenji Mizuguchi,Eng Eong Ooi

doi:10.1371/journal.pone.0060038

Abstract

5,6-Dimethylxanthenone-4-acetic acid (DMXAA), a potent type I interferon (IFN) inducer, was evaluated as a chemotherapeutic agent in mouse cancer models and proved to be well tolerated in human cancer clinical trials. Despite its multiple biological functions, DMXAA has not been fully characterized for the potential application as a vaccine adjuvant. In this report, we show that DMXAA does act as an adjuvant due to its unique property as a soluble innate immune activator. Using OVA as a model antigen, DMXAA was demonstrated to improve on the antigen specific immune responses and induce a preferential Th2 (Type-2) response. The adjuvant effect was directly dependent on the IRF3-mediated production of type-I-interferon, but not IL-33. DMXAA could also enhance the immunogenicity of influenza split vaccine which led to significant increase in protective responses against live influenza virus challenge in mice compared to split vaccine alone. We propose that DMXAA can be used as an adjuvant that targets a specific innate immune signaling pathway via IRF3 for potential applications including vaccines against influenza which requires a high safety profile.

Highlights

Dimethylxanthenone-4-acetic acid (DMXAA) was developed as a vascular disruptive agent for use in cancer therapy
We found that DMXAA could significantly augment specific immune responses against OVA, as indicated by the increase in serum anti-OVA total IgG titers compared to OVA alone immunized group (Figure 1A)
No T-cell responses could be detected in OVA plus Alum or DMXAA groups, whereas splenocytes from the OVA plus CpG group responded with high IFN-c secretion in the presence CD8 peptide and whole OVA protein (Figure 1F)

Summary

Introduction

DMXAA was developed as a vascular disruptive agent for use in cancer therapy. Several clinical trials, including a recently completed phase III clinical trial for non-small cell lung carcinoma, have shown that DMXAA is safe and well-tolerated in humans [1]. The anti-neoplastic property of DMXAA is largely attributed to its induction of TNFa which can be detected in the serum and tumor microenvironment within hours of administration [3] It can activate several inflammatory cell signaling pathways, including extracellular signal-regulated kinases 1 and 2, c-Jun N-terminal kinases, and cytosolic nucleotide-binding oligomerization domain 1 and 2like receptors [4,5]. DMXAA resembles viral infections and double stranded DNA (dsDNA) in the inflammatory signaling events it triggers to induce type-I-IFN production [8]. It utilizes the TBK1-IRF3 signaling pathway without the involvement of Toll-like receptors (TLRs) or RNA helicases for its mechanism of type-I-IFN induction. Due to its ability to induce strong typeI-IFN, DMXAA was found to be an effective antiviral agent against influenza [14,15]

Methods

Results

Conclusion