Serum Proteome and Cytokine Analysis in a Longitudinal Cohort of Adults with Primary Dengue Infection Reveals Predictive Markers of DHF

Yadunanda Kumar,Zheng Bo,Eng Eong Ooi,Steven R Tannenbaum,Jagath C Rajapakse,Cui Liang,Alan L Rothman

doi:10.1371/journal.pntd.0001887

Abstract

BackgroundInfections caused by dengue virus are a major cause of morbidity and mortality in tropical and subtropical regions of the world. Factors that control transition from mild forms of disease such as dengue fever (DF) to more life-threatening forms such as dengue hemorrhagic fever (DHF) are poorly understood. Consequently, there are no reliable methods currently available for early triage of DHF patients resulting in significant over-hospitalization.Methodology/Principal FindingsWe have systematically examined the proteome, cytokines and inflammatory markers in sera from 62 adult dengue patients (44 DF; 18 DHF) with primary DENV infection, at three different times of infection representing the early febrile, defervescence and convalescent stages. Using fluorescent bioplex assays, we measured 27 cytokines in these serum samples. Additionally, we used multiple mass spectrometry methods for iTRAQ-based comparative analysis of serum proteome as well as measurements of protein adducts- 3-nitrotyrosine and 3-chlorotyrosine as surrogate measures of free radical activity. Using multiple methods such as OPLS, MRMR and MSVM-RFE for multivariate feature selection and classification, we report molecular markers that allow prediction of primary DHF with sensitivity and specificity of >80%.Conclusions/SignificanceThis report constitutes a comprehensive analysis of molecular signatures of dengue disease progression and will help unravel mechanisms of dengue disease progression. Our analysis resulted in the identification of markers that may be useful for early prediction of DHF during the febrile phase. The combination of highly sensitive analytical methods and novel statistical approaches described here forms a robust platform for biomarker discovery.

Highlights

Infection with dengue virus (DENV) causes a spectrum of clinical manifestations ranging from mild dengue fever (DF) to the potentially lethal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) [1]
We have further developed novel statistical methods that allow us to identify small panels of measureable blood markers, which can distinguish dengue patients that develop milder, self-limiting form of the disease from those that progress to develop severe symptoms
Because these markers can be applied within 48–72 hours of onset of febrile symptoms, we expect them to be useful for early classification of severe dengue disease

Summary

Introduction

Infection with dengue virus (DENV) causes a spectrum of clinical manifestations ranging from mild dengue fever (DF) to the potentially lethal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) [1]. Dengue is endemic to the tropical and sub-tropical regions of the world, which are home to over half the population of the world as well as being popular tourist destinations It has emerged in new areas such as south Florida and Mediterranean France. Recognition of patients with plasma leakage is critical for the initiation of appropriate fluid management to prevent onset of hypovolemic shock. Because these symptoms become evident only in the critical phase of infection, it is currently not possible to distinguish DF and DHF accurately during the early stages of illness, when the disease is less well differentiated [3]. There are no reliable methods currently available for early triage of DHF patients resulting in significant over-hospitalization

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Conclusion