Assessing Host Immune Response to Dengue Virus Infection at Single‐Cell Resolution

Abstract

Dengue virus (DENV) is a mosquito‐borne pathogen, commonly found in tropical and subtropical climates, that results in a broad range of disease pathologies ranging from asymptomatic to dengue fever (DF) to the more severe dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Despite reports characterizing elevated levels of inflammatory cytokines in serum and correlating the manifestation of the severe disease symptoms to increased immune responses, attempts to delineate the signaling profile have been hindered by the low‐efficiency, dose‐dependent DENV infection of cultured cells. Understanding the signaling profile may reveal the hallmarks of disease progression and dictate the manifested severity of the infection. In this study, we eliminate the complexity of interpreting averaged signaling profile from heterogeneous population by isolating infected cells in a single‐cell barcode chip (SCBC), a device that enable simultaneous detection of multiple secreted cytokines at the single‐cell level. This strategy will allow us to effectively differentiate DENV‐infected from non‐infected cells, profile the types and levels of cytokines and chemokines secreted by these infected cells, and examine how various immune cells response to DENV infection. This single‐cell signaling profile will provide insight into how DENV infection may perturb the host immune system signaling networks and aid in the identification of therapeutic targets that can modulate DENV infection and suppress progression into DHF/DSS.Supported by NIH grant U19‐AI089992