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Plugging the Leak in Dengue Shock

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Dengue and Zika: Control and Antiviral Treatment Strategies

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1062))

Abstract

Recent structural and functional advances provide fresh insight into the biology of the dengue virus non-structural protein, NS1 and suggest new avenues of research. The work of our lab and others have shown that the secreted, hexameric form of NS1 has a systemic toxic effect, inducing inflammatory cytokines and acting directly on endothelial cells to produce the hallmark of dengue disease, vascular leak. We also demonstrated that NS1 exerts its toxic activity through recognition by the innate immune receptor TLR4, mimicking the bacterial endotoxin LPS. This monograph covers the background underpinning these new findings and discusses new avenues for antiviral and vaccine intervention.

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Authors and Affiliations

  1. Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, Australia

    Daniel Watterson, Naphak Modhiran, David A. Muller, Katryn J. Stacey & Paul R. Young

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Editors and Affiliations

  1. Institute of Biochemistry, University of Lübeck, Lübeck, Germany

    Rolf Hilgenfeld

  2. Emerging Infectious Diseases Program, Duke-NUS Medical School Singapore, Singapore, Singapore

    Subhash G. Vasudevan

Discussion of Chapter 7 in Dengue and Zika: Control and Antiviral Treatment Strategies

Discussion of Chapter 7 in Dengue and Zika: Control and Antiviral Treatment Strategies

This discussion was held at the 2nd Advanced Study Week on Emerging Viral Diseases at Praia do Tofo, Mozambique.Transcribed by Hilgenfeld R and Vasudevan SG (Eds); approved by Dr. Paul Young.

  • Subhash Vasudevan: Which Virus strains did you use for the mouse challenge experiment?

  • Paul Young: We have done two principle studies, multiple times. S221 which is the Shresta mouse-adapted strain in AG129. And also an East Timor clinical strain, not mouse-adapted for DENV1.

  • George Gao: Have you compared the dimer versus the hexamer in your experiment?

  • Paul Young: You can’t use the dimer form in tissue culture experiments. The dimer in a non-detergent enviroment would aggregate. That was one of the biggest issues in terms of trying to crystallize it. It was crystallized in the end in the presence of detergent, so you can break down the hexamer into a dimer, but then it aggregates again when you put it into a normal detergent-free, in vivo enviroment.

  • George Gao: So then the question is that both Janet Smith and my group have published NS1 dimer structures. We think it is a dimer.

  • Paul Young: Yes, you got the Zika NS1 dimer but I would assume there is detergent in the crystallization buffer.

  • George Gao: But can you purify it?

  • Paul Young: If you purify it in the context of detergent you get the dimer form. We routinely purify it as a hexamer in the absence of detergent. So what I am saying is: you can purify it in its dimer form, and can maintain it as a dimer when there is detergent present. But if you put it into an in vivo system, if you take away the detergent it aggregates because of the exposed hydrophobic domains.

  • Norbert Heinrich: This may be a naive question, but seeing as you showed that NS1 triggers IL6 release from PBMC, obviously the next step would be CRP (C-reactive protein) production in the liver. Do you know whether that is clinically relevant following the conclusion that we can not use CRP to discriminate viral from bacterial infections in Dengue?

  • Paul Young: That could be true. Hepatomegaly is a significant clinical feature for Dengue patients. So we have seen that is happening and CRP is shown to be elevated in Dengue patients.

  • Norbert Heinrich: And likewise would you think the same is true for the other Flaviviruses and their NS1?

  • Paul Young: I never try to extrapolate from one flavivirus to another Aravinda. Hepatomegaly is a feature of other flaviviruses and maybe it is for yellow fever?

  • Aravinda de Silva: Yes. I do not know.

  • Paul Young: Yes I do not know either. It is possible.

  • Aravinda de Silva: So how do you explain the reduced viremia which is a very surprising result?

  • Paul Young: We were also surprised. What we do know – there has been some literature, and we have confirmed it ourselves – is that if you add NS1 to macrophages that have been infected you will increase viral infection. We think that it is simply because the activated profile of the macrophages are a much better enviroment for the virus to grow. So you can imagine that knocking down that effect might have some impact, but we didn’t expect to knock it back completely. I can imagine that some drop in viral load, but not completely. We are currently trying to investigate the mechanism.

  • Subhash Vasudevan: And you see this 48 hours post-infection which is spectacular.

  • Paul Young: Yes and also the fact that treatment at day zero would be exactly the wrong thing to be doing.

  • Shi Yi: Do you have any direct binding data for toll-like receptor 4 and NS1?

  • Paul Young: We have co-precipitation data. What we are obviously trying to do is to get a co-crystal structure. That is going to be a huge challenge, but we are doing the co-precipitation studies and have shown that it will co-precipitate when you combine them.

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Watterson, D., Modhiran, N., Muller, D.A., Stacey, K.J., Young, P.R. (2018). Plugging the Leak in Dengue Shock. In: Hilgenfeld, R., Vasudevan, S. (eds) Dengue and Zika: Control and Antiviral Treatment Strategies. Advances in Experimental Medicine and Biology, vol 1062. Springer, Singapore. https://doi.org/10.1007/978-981-10-8727-1_7

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