Quantification of the frequency and multiplicity of infection of respiratory- and lymph node-resident dendritic cells during influenza virus infection

Rebecca VanoOsten Anderson; Jodi McGill; Kevin L Legge

doi:10.1371/journal.pone.0012902

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Quantification of the frequency and multiplicity of infection of respiratory- and lymph node-resident dendritic cells during influenza virus infection

Journal article

Open access

Peer reviewed

Quantification of the frequency and multiplicity of infection of respiratory- and lymph node-resident dendritic cells during influenza virus infection

Rebecca VanoOsten Anderson, Jodi McGill and Kevin L Legge

PloS one, Vol.5(9), pp.e12902-e12902

09/23/2010

DOI: 10.1371/journal.pone.0012902

PMCID: PMC2944834

PMID: 20886117

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Abstract

Previous studies have demonstrated that DC differentially regulate influenza A virus (IAV)-specific CD8 T cell responses in vivo during high and low dose IAV infections. Furthermore, in vitro infection of DC with IAV at low versus high multiplicities of infection (MOI) results in altered cytokine production and a reduced ability to prime naïve CD8 T cell responses. Flow cytometric detection of IAV proteins within DC, a commonly used method for detection of cellular IAV infection, does not distinguish between the direct infection of these cells or their uptake of viral proteins from dying epithelial cells. We have developed a novel, sensitive, single-cell RT-PCR-based approach to assess the infection of respiratory DC (rDC) and lymph node (LN)-resident DC (LNDC) following high and low dose IAV infections. Our results show that, while a fraction of both rDC and LNDC contain viral mRNA following IAV infection, there is little correlation between the percentage of rDC containing viral mRNA and the initial IAV inoculum dose. Instead, increasing IAV inoculums correlate with augmented rDC MOI. Together, our results demonstrate a novel and sensitive method for the detection of direct IAV infection at the single-cell level and suggest that the previously described ability of DC to differentially regulate IAV-specific T cell responses during high and low dose IAV infections could relate to the MOI of rDC within the LN rather than the percentage of rDC infected.

Cell Line

Influenza A virus - physiology

Dendritic Cells - immunology

Humans

Influenza A virus - genetics

Viral Proteins - genetics

Influenza A virus - isolation & purification

Lymph Nodes - virology

Lymph Nodes - immunology

Respiratory System - virology

Animals

Reverse Transcriptase Polymerase Chain Reaction - methods

Female

Influenza A virus - immunology

Influenza, Human - virology

Mice

Mice, Inbred BALB C

Dendritic Cells - virology

Respiratory System - immunology

Influenza, Human - immunology

Details

Title: Subtitle: Quantification of the frequency and multiplicity of infection of respiratory- and lymph node-resident dendritic cells during influenza virus infection
Creators: Rebecca VanoOsten Anderson - Department of Pathology, Interdisciplinary Graduate Program in Immunology, University of Iowa, Iowa City, Iowa, United States of America
Jodi McGill
Kevin L Legge
Resource Type: Journal article
Publication Details: PloS one, Vol.5(9), pp.e12902-e12902
DOI: 10.1371/journal.pone.0012902
PMID: 20886117
PMCID: PMC2944834
NLM abbreviation: PLoS One
ISSN: 1932-6203
eISSN: 1932-6203
Publisher: Public Library of Science; United States
Grant note: R01 AI071085 / NIAID NIH HHS AI-071085 / NIAID NIH HHS R56 AI071085 / NIAID NIH HHS
Language: English
Date published: 09/23/2010
Academic Unit: Microbiology and Immunology; Pathology
Record Identifier: 9984047698602771

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