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. 2017 Aug 24;91(18):e00617-17.
doi: 10.1128/JVI.00617-17. Print 2017 Sep 15.

Transcriptional Profiling Confirms the Therapeutic Effects of Mast Cell Stabilization in a Dengue Disease Model

Juliet Morrison  1 , Abhay P S Rathore  2   3 , Chinmay K Mantri  2 , Siti A B Aman  2 , Andrew Nishida  4 , Ashley L St John  5   3   6
Affiliations

Transcriptional Profiling Confirms the Therapeutic Effects of Mast Cell Stabilization in a Dengue Disease Model

Juliet Morrison et al. J Virol. .

Abstract

There are no approved therapeutics for the treatment of dengue disease despite the global prevalence of dengue virus (DENV) and its mosquito vectors. DENV infections can lead to vascular complications, hemorrhage, and shock due to the ability of DENV to infect a variety of immune and nonimmune cell populations. Increasingly, studies have implicated the host response as a major contributor to severe disease. Inflammatory products of various cell types, including responding T cells, mast cells (MCs), and infected monocytes, can contribute to immune pathology. In this study, we show that the host response to DENV infection in immunocompetent mice recapitulates transcriptional changes that have been described in human studies. We found that DENV infection strongly induced metabolic dysregulation, complement signaling, and inflammation. DENV also affected the immune cell content of the spleen and liver, enhancing NK, NKT, and CD8+ T cell activation. The MC-stabilizing drug ketotifen reversed many of these responses without suppressing memory T cell formation and induced additional changes in the transcriptome and immune cell composition of the spleen, consistent with reduced inflammation. This study provides a global transcriptional map of immune activation in DENV target organs of an immunocompetent host and supports the further development of targeted immunomodulatory strategies to treat DENV disease.IMPORTANCE Dengue virus (DENV), which causes febrile illness, is transmitted by mosquito vectors throughout tropical and subtropical regions of the world. Symptoms of DENV infection involve damage to blood vessels and, in rare cases, hemorrhage and shock. Currently, there are no targeted therapies to treat DENV infection, but it is thought that drugs that target the host immune response may be effective in limiting symptoms that result from excessive inflammation. In this study, we measured the host transcriptional response to infection in multiple DENV target organs using a mouse model of disease. We found that DENV infection induced metabolic dysregulation and inflammatory responses and affected the immune cell content of the spleen and liver. The use of the mast cell stabilization drug ketotifen reversed many of these responses and induced additional changes in the transcriptome and immune cell repertoire that contribute to decreased dengue disease.

Keywords: dengue fever; dengue virus; interferons; mast cell; transcriptional regulation.

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Figures

FIG 1
FIG 1
Metabolic dysregulation and inflammatory cytokine signaling dominate the host response to DENV2 infection. (A and B) Numbers of DE genes in DENV-infected livers and spleens on day 1 (A) and day 3 (B). (C) Day 1 network of DE interferon response and metabolism genes in the liver. (D and E) Network of immune cell activation, interferon response, and cytokine response genes in the spleen on day 1 (D) and day 3 (E) post-DENV infection. Criteria used for differential expression analysis were an adjusted P value of <0.05, as determined by the limma empirical Bayes-moderated t test, and a log2 FC of >0.58. Blue represents downregulation, red represents upregulation, and gray represents no differential expression.
FIG 2
FIG 2
Ketotifen treatment induces a robust host response in DENV2-infected livers and spleens. (A and B) DE genes in DENV-infected and ketotifen-treated organs on day 1 (A) and on day 3 (B). (C to F) Overlaps in the numbers of DE genes between DENV-infected and DENV-infected, ketotifen-treated samples from liver (C and E) and spleen (D and F) on day 1 (C and D) and day 3 (E and F). Criteria used for differential expression analysis were an adjusted P value of <0.05, as determined by the limma empirical Bayes-moderated t test, and a log2 FC of >0.58.
FIG 3
FIG 3
Ketotifen perturbs multiple pathways in the liver. The liver DE genes from Fig. 1 and 2 were analyzed by IPA to produce lists of host pathways that were most perturbed by DENV infection with or without ketotifen treatment. (A and B) The top 10 enriched pathways on day 1 (A) and day 3 (B) are represented as radial plots. The distance from the center in each radial plot represents the enrichment score, which is defined as −log10(P value), using a right-tailed Fisher exact test. (C) IPA network showing interactions between molecules involved in cholesterol biosynthesis and those involved in intrinsic prothrombin activation on day 1. Blue represents downregulation, red represents upregulation, and gray represents no differential expression.
FIG 4
FIG 4
Ketotifen perturbs multiple pathways in the spleen. The spleen DE genes from Fig. 1 and 2 were analyzed by IPA to produce lists of host pathways that were most perturbed by DENV infection with or without ketotifen treatment. (A and B) The top 10 enriched pathways at day 1 (A) and day 3 (B) are represented as radial plots. The distance from the center in each radial plot represents the enrichment score, which is defined as −log10(P value), using a right-tailed Fisher exact test. PRRs, pattern recognition receptors. (C) IPA network showing interactions between molecules involved in cholesterol biosynthesis, complement signaling, lipid metabolism, and cross talk between immune cells on day 1. Blue represents downregulation, red represents upregulation, and gray represents no differential expression.
FIG 5
FIG 5
Ketotifen treatment is predicted to modulate lymphocyte numbers in the spleen. The tissue deconvolution-based DCQ algorithm was used to infer immune cell quantities from gene expression changes in the spleen. Enriched natural killer (A), NKT (B), and CD8+ (C) cell populations in spleens of mice infected with DENV2 with or without ketotifen treatment are presented as heat maps.
FIG 6
FIG 6
Flow cytometry validates DCQ predictions. Flow cytometry was performed to validate that DCQ correctly interpreted the frequencies of lymphocytes that were differentially enriched or activated during DENV infection and/or ketotifen treatment. (A) Representative plots from day 3 showing the percentages of NK and NKT cells in the spleen. (B) Quantitation of NKT cells from panel A. (C) Representative plots showing the quantities of CD44+ NKT cells on day 3. (D) Percent expression of CD44 on NKT cells corresponding to data shown in panel C (n = 5). (E) Representative plots showing the percentages of NK cells that are CD4+ on day 1. (F) Quantitation of activated NK cells (NK1.1+ CD4+) on days 1 and 3. (G) Representative plots from day 3 showing the percentages of CD8+ T cells in the spleen. (H) Quantitation of CD8+ T cells from panel G (n = 5). For all panels containing graphs (n = 5), analysis was performed by 2-way analysis of variance (* indicates a P value of <0.05).
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References

    1. WHO. 2016. Dengue and severe dengue. WHO fact sheet. WHO, Geneva, Switzerland.
    1. Nishiura H, Halstead SB. 2007. Natural history of dengue virus (DENV)-1 and DENV-4 infections: reanalysis of classic studies. J Infect Dis 195:1007–1013. doi:10.1086/511825. - DOI - PubMed
    1. Bustos-Arriaga J, Garcia-Machorro J, Leon-Juarez M, Garcia-Cordero J, Santos-Argumedo L, Flores-Romo L, Mendez-Cruz AR, Juarez-Delgado FJ, Cedillo-Barron L. 2011. Activation of the innate immune response against DENV in normal non-transformed human fibroblasts. PLoS Negl Trop Dis 5:e1420. doi:10.1371/journal.pntd.0001420. - DOI - PMC - PubMed
    1. Dalrymple NA, Mackow ER. 2012. Endothelial cells elicit immune-enhancing responses to dengue virus infection. J Virol 86:6408–6415. doi:10.1128/JVI.00213-12. - DOI - PMC - PubMed
    1. Kurane I, Janus J, Ennis FA. 1992. Dengue virus infection of human skin fibroblasts in vitro production of IFN-beta, IL-6 and GM-CSF. Arch Virol 124:21–30. doi:10.1007/BF01314622. - DOI - PubMed

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