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An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection

An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection

Published on: December 10, 2013

Relative respiratory syncytial virus cytopathogenesis in upper and lower respiratory tract epithelium

Hong Guo-Parke¹, Paul Canning, Isobel Douglas

¹1 Centre for Infection and Immunity, School of Medicine, Dentistry and Biomedical Sciences, Queens University Belfast, Belfast, Northern Ireland, United Kingdom.

American Journal of Respiratory and Critical Care Medicine

|August 20, 2013

Insights

Respiratory syncytial virus (RSV) causes similar, though less severe, damage in infant nasal and bronchial cells. Nasal cells offer a viable model for studying RSV pathogenesis in infants.

Area of Science:

Virology
Immunology
Pediatrics

Background:

Respiratory syncytial virus (RSV) is a common pathogen affecting infant airways.
While most infants experience mild symptoms, a significant portion develops lower respiratory tract (LRT) involvement.
The precise mechanisms of RSV cytopathogenesis in infant upper respiratory tract (URT) versus LRT remain unclear.

Purpose of the Study:

To compare the cytopathogenesis of RSV infection in infant nasal and bronchial epithelial cells.
To utilize novel models of well-differentiated primary pediatric nasal epithelial cells (WD-PNECs) and bronchial epithelial cells (WD-PBECs).

Main Methods:

Generated WD-PNECs and WD-PBECs from pediatric nasal and bronchial brushes.
Infected cells with RSV BT2a and assessed tropism, infectivity, cytopathology, and viral growth kinetics.
Measured cell sloughing, apoptosis, and inflammatory cytokine/chemokine responses.

Main Results:

RSV primarily infected ciliated cells in both nasal and bronchial cultures, without significant gross cytopathology.
Higher RSV growth kinetics and peak titers were observed in WD-PBECs compared to WD-PNECs.
Both cell types exhibited increased cell sloughing, apoptosis, and induction of lambda IFNs, with similar chemokine responses.

Conclusions:

RSV induces comparable cytopathogenesis and inflammatory responses in infant nasal and bronchial epithelial cells, with quantitative differences.
WD-PNECs serve as a valid surrogate model for studying RSV pathogenesis in infant airway epithelium.
Findings contribute to understanding RSV disease progression from URT to LRT in infants.

Read full analysis

Abstract

Rationale:

Respiratory syncytial virus (RSV) is a major pathogen that primarily infects airway epithelium. Most infants suffer mild upper respiratory tract (URT) symptoms, whereas approximately one-third progress to lower respiratory tract (LRT) involvement. Despite the ubiquity of URT infection, little is known about the relative cytopathogenesis of RSV infection in infant URT and LRT.

Objectives:

This study aimed to compare RSV cytopathogenesis in nasal- and bronchial-derived epithelium from the same individuals using novel models derived from well-differentiated primary pediatric nasal (WD-PNECs) and bronchial epithelial cells (WD-PBECs).

Methods:

WD-PNECs and WD-PBECs were generated from nasal and bronchial brushes, respectively, and mock-infected or infected with RSV BT2a. RSV tropism, infectivity, cytopathology, growth kinetics, cell sloughing, apoptosis, and cytokine and chemokine responses were determined.

Measurements And Main Results:

RSV infection in both cultures was restricted to apical ciliated cells and occasional nonciliated cells but not goblet cells. It did not cause gross cytopathology. Infection resulted in apical release of progeny virus, increased apical cell sloughing, apoptosis, and occasional syncytia. RSV growth kinetics and peak titers were higher in WD-PBECs, coincident with higher ciliated cell contents, cell sloughing, and slightly compromised tight junctions. However, proinflammatory chemokine responses were similar for both cultures. Also, lambda IFNs, especially IL-29, were induced by RSV infection.

Conclusions:

RSV induced remarkably similar, albeit quantitatively lower, cytopathogenesis and proinflammatory responses in WD-PNECs compared with WD-PBECs that reproduce many hallmarks of RSV pathogenesis in infants. WD-PNECs may provide an authentic surrogate model with which to study RSV cytopathogenesis in infant airway epithelium.

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