Dengue Virus Targets and Efficiently Replicates in Megakaryocytes Open Access

Abstract

The vectorborne pathogen dengue virus (DENV) infects millions of persons worldwide and can be lethal in the young and the old. In spite of decades of research, the primary cellular target responsible for causing high DENV viremia in humans remains elusive. Several cell lineages (dendritic cells, macrophages/monocytes, B lymphocytes, endothelial cells, and megakaryocyte- erythrocyte progenitor cells) have been implicated as targets and have been linked to different pathomechanisms. The widely recognized clinical findings of thrombocytopenia and coagulopathy observed in dengue patients directed our investigations towards the relationship between DENV and megakaryocytes, the platelet progenitors. We have examined the permissiveness of megakaryocyte- erythrocyte progenitor (MEPs) cell lines, primary rhesus macaque bone marrow cultures, and primary human bone marrow tissue and have found they are readily susceptible to DENV serotype 2 (DENV2) infection leading to productive replication. Sequential analyses of bone marrow samples from infected monkeys showed that DENV2 viral antigens were included in multinucleated cells that expressed CD61 and CD41a early post infection, at times corresponding with peak viral titer, suggesting that megakaryocytes are a target cell contributing to viremia. Of interest, after the viral peak, DENV2 antigen gradually shifted to monocyte/macrophage cells, suggesting these cells play a role later in infection not critical for initial viremia. These results have important implications for the development of dengue antivirals, the generation of effective vaccines, and the safety of blood and platelet donations in DENV endemic areas.

Table of Contents

Abstract. iv

Acknowledgements. v

Table of Contents. vi

List of Figures and Tables. viii

CHAPTER 1 - Introduction. 1

DENGUE VIRUS GENERAL BACKGROUND. 1

VIRAL TARGETS. 3

VIRUS ASSEMBLY AND STRUCTURE. 6

DENGUE DISEASE. 15

DENGUE DISEASE MECHANISMS. 20

TREATMENT AND PREVENTION. 25

CHAPTER 2 - Characterization of dengue virus 2 growth in megakaryocyte-erythrocyte progenitor Cells. 31

ABSTRACT. 33

INTRODUCTION. 34

RESULTS. 36

DISCUSSION. 56

MATERIALS AND METHODS. 58

REFERENCES. 65

CHAPTER 3 - Multiploid CD61+ cells are the pre-dominant cell lineage infected during acute dengue virus infection in bone marrow. 73

ABSTRACT. 75

INTRODUCTION. 76

METHODS. 78

RESULTS. 84

DISCUSSION.110

REFERENCES. 115

CHAPTER 4 - Role of microparticles in dengue virus infection and its impact of medical intervenetion strategies. 120

ABSTRACT. 122

INTRODUCTION. 123

DENGUE VIRUS'S PROPAGATION AND STRUCTURE IN VIVO AND IN VITRO. 124

VIRUSES OF ALTERNATE MORPHOLOGY (VAMS). 126

MICROPARTICLES (MPS) AND THEIR INVOLVEMENT IN INFECTIONS. 131

VECTOR-BORNE DISEASE TRANSMISSION. 135

IMPLICATIONS AND CURRENT DENGUE VACCINE EFFORTS. 136

IMPLICATIONS ON DRUG DESIGN. 139

SUMMARY AND CONCLUSIONS. 142

REFERENCES. 143

CHAPTER 5 - Can nonhuman primates serve as models for investigating dengue disease pathogenesis?. 161

ABSTRACT. 163

INTRODUCTION. 164

DEVELOPMENT OF DENV INFECTION ANIMAL MODEL SYSTEM. 166

MOUSE MODEL. 167

NONHUMAN PRIMATE (NHP) MODELS. 168

VIRUS DELIVERY. 180

RHESUS MACAQUE MODEL OF COAGULOPATHY. 181

BONE MARROW (BM) TARGETING. 184

PLATELET ACTIVITIES. 187

POTENTIAL REFINEMENTS TO THE COAGULOPATHY MONKEY MODEL. 190

HOST CHARACTERISTICS OR GENETIC FACTORS THAT INCREASE SUSCEPTIBILITY TO COAGULOPATHY. 192

CONCLUSIONS. 193

REFERENCES. 195

CHAPTER 6 - Discussion and future directions. 213

REFERENCES. 224

About this Dissertation

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School	Laney Graduate School
Department	Biological and Biomedical Sciences
Subfield / Discipline	Microbiology & Molecular Genetics
Degree	PhD
Submission	Dissertation
Language	English
Research Field	Health Sciences, Pathology Biology, Virology Biology, Microbiology
Keyword	Megakaryocytes Dengue Virus Rhesus Macaque Megakaryocyte-Erythrocyte Progenitor
Committee Chair / Thesis Advisor	Villinger, Francois, Emory University Perng, Guey Chuen 'Oscar', Emory University
Committee Members	Grakoui, Arash, Emory University Steinhauer, David, Emory University Wright, Elizabeth R., Emory University Anderson, Larry J, Emory University

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