High-dimensional CyTOF analysis of dengue virus-infected human DCs reveals distinct viral signatures.

Rebecca E Hamlin,Ana Fernandez-Sesma,El-Ad D Amir,Kevin Maringer,Adeeb Rahman,Ana M Maestre,Andrew Kasarskis,Anthony C Fredericks,Theodore R Pak,Dabeiba Bernal-Rubio,Miriam Merad,Uma Potla,Ignacio Mena,Irene Ramos

doi:10.1172/jci.insight.92424

Rebecca E Hamlin, Ana Fernandez-Sesma + Show 12 more

Open Access

https://doi.org/10.1172/jci.insight.92424

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Abstract

Dengue virus (DENV) is the most prevalent mosquito-borne virus causing human disease. Of the 4 DENV serotypes, epidemiological data suggest that DENV-2 secondary infections are associated with more severe disease than DENV-4 infections. Mass cytometry by time-of-flight (CyTOF) was used to dissect immune changes induced by DENV-2 and DENV-4 in human DCs, the initial targets of primary infections that likely affect infection outcomes. Strikingly, DENV-4 replication peaked earlier and promoted stronger innate immune responses, with increased expression of DC activation and migration markers and increased cytokine production, compared with DENV-2. In addition, infected DCs produced higher levels of inflammatory cytokines compared with bystander DCs, which mainly produced IFN-induced cytokines. These high-dimensional analyses during DENV-2 and DENV-4 infections revealed distinct viral signatures marked by different replication strategies and antiviral innate immune induction in DCs, which may result in different viral fitness, transmission, and pathogenesis.

Highlights

Dengue virus (DENV) is the most prevalent mosquito-borne virus causing disease in humans [1], with an estimated 390 million annual infections worldwide, of which 96 million manifest with clinical symptoms [2]
Using cytometry by time-of-flight (CyTOF) analysis to measure intracellular DENV structural and nonstructural proteins and titration of extracellular infectious DENV particles, we found that DENV-2 peak infection occurred at 48 hours after infection, whereas DENV-4 infection peaked at 24 hpi and was significantly diminished by 72 hpi (Figure 1, A–D)
Even though DENV-2 infected a smaller percentage of cells compared with DENV-4, those DCs infected with DENV-2 showed significantly higher levels of DENV nonstructural 3 (NS3) protein (Figure 1C) at 48 and 72 hpi compared with the DCs infected with DENV-4

Summary

Introduction

Dengue virus (DENV) is the most prevalent mosquito-borne virus causing disease in humans [1], with an estimated 390 million annual infections worldwide, of which 96 million manifest with clinical symptoms [2]. Dengue disease may present as a nonspecific febrile illness or as a more severe infection marked by hemorrhage or circulatory failure [3, 4]. DENV is a flavivirus that exists as 4 distinct serotypes (DENV-1– DENV-4) and is transmitted by Aedes mosquitoes [5]. At the amino acid level, the 4 DENV serotypes share limited similarity among serotypes (about 60%–75%) but about 97% similarity within a given serotype [6]. Epidemiological studies have reported differences between serotypes with regard to disease severity. DENV-2 secondary infections have been associated with more severe disease, while DENV-4 infections have been correlated with more mild disease [7,8,9,10,11,12]

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