High-dimensional CyTOF analysis of dengue virus–infected human DCs reveals distinct viral signatures
Rebecca E. Hamlin, Adeeb Rahman, Theodore R. Pak, Kevin Maringer, Ignacio Mena, Dabeiba Bernal-Rubio, Uma Potla, Ana M. Maestre, Anthony C. Fredericks, El-ad D. Amir, Andrew Kasarskis, Irene Ramos, Miriam Merad, Ana Fernandez-Sesma
Rebecca E. Hamlin, Adeeb Rahman, Theodore R. Pak, Kevin Maringer, Ignacio Mena, Dabeiba Bernal-Rubio, Uma Potla, Ana M. Maestre, Anthony C. Fredericks, El-ad D. Amir, Andrew Kasarskis, Irene Ramos, Miriam Merad, Ana Fernandez-Sesma
View: Text | PDF
Research Article Immunology Virology

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

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.

Authors

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

×

Figure 5

Bystander cells show distinct cytokine and cleaved caspase-3 phenotypes compared with infected cells during DENV infection.

Options: View larger image (or click on image) Download as PowerPoint
Bystander cells show distinct cytokine and cleaved caspase-3 phenotypes ...
Dengue virus–infected (DENV-infected) DCs were gated on live single CD45+ cells, as shown in Supplemental Figure 3A. Using mass cytometry by time-of-flight (CyTOF) analysis for each DENV-2 or DENV-4 sample, the cells were stratified as DENV-infected (I) cells (filled circles) or bystander (B) cells (open circles). Mock-infected cells (M) were analyzed as a single population. Seven DC donors are shown. The percentage of mock, bystander, or DENV-infected cells of DENV-2 (green) and DENV-4 (purple) samples that stained positive intracellularly for the cytokines (A) TNF-α, (B) IL-1β, (C) IL-8, and (D) IP-10 and for the apoptosis marker (E) cleaved caspase-3 are shown. Mean ± SEM is shown. Black asterisks represent statistical significance by the paired, 2-tailed Wilcoxon signed-rank test, comparing bystander and infected cells within a given culture sample at a given time point. The Benjamini-Hochberg procedure was performed within each time point to adjust the significance level for multiple comparisons (*P ≤ 0.05).