Apoptosis induced by Pseudomonas aeruginosa in antigen presenting cells is diminished by genetic modification with CD40 ligand

Stefan Worgall, Katherine Martushova, Annette Busch, Leah Lande, Ronald Crystal

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Persistent colonization with Pseudomonas aeruginosa (PA) is a hallmark of the lung disease associated with cystic fibrosis (CF). Based on the concept that PA is not cleared from the lung by the host response in individuals with CF, we analyzed the capacity of PA to induce cell death in human alveolar macrophages (AM) and murine dendritic cells (DC), antigen presenting cells that play a central role in the initiation of pulmonary host defenses against pathogens, and evaluated if genetic modification can lead to protection against PA induced cell death. AM and DC were susceptible to cell death induced by the laboratory PA isolates PAO1, PAK and PA103, as well as a mucoid derivative of PAO1 and PA isolates derived from sputum of individuals with CF. Apoptosis, analyzed by TUNEL assay, was detectable in AM and DC as early as 3 h after infection with PA. In contrast, the same strains and doses of PA had little effect on the lung epithelial cell line A549 and primary cultures of human bronchial epithelial cells in vitro. Pretreatment of DC with the caspase inhibitors VAD-fmk and YVAD-cmk reduced PA induced cell death (p < 0.05). Finally, genetic modification of DC to express CD40L using an adenovirus vector decreased the susceptibility of DC to cell death induced by PAO1 compared with DC infected with a control Ad vector (p < 0.01). The data demonstrate that DC and AM are susceptible to apoptosis induced by PA and that this response can be partially reversed by genetic modification with CD40L, a CD4+ T cell molecule that plays a central role in activating antigen presenting cells. These observations suggest a potential mechanism contributing to the persistence of PA in CF and suggest that genetic manipulation of antigen presenting cells with anti-apoptotic genes may be able to strengthen host defenses in CF.

Original languageEnglish
Pages (from-to)636-644
Number of pages9
JournalPediatric Research
Volume52
Issue number5
DOIs
Publication statusPublished - 1 Nov 2002
Externally publishedYes

Fingerprint

CD40 Ligand
Antigen-Presenting Cells
Pseudomonas aeruginosa
Apoptosis
Dendritic Cells
Cystic Fibrosis
Alveolar Macrophages
Cell Death
Alveolar Epithelial Cells
Lung
Epithelial Cells
Caspase Inhibitors
In Situ Nick-End Labeling
Sputum
Adenoviridae
Lung Diseases

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

Cite this

Apoptosis induced by Pseudomonas aeruginosa in antigen presenting cells is diminished by genetic modification with CD40 ligand. / Worgall, Stefan; Martushova, Katherine; Busch, Annette; Lande, Leah; Crystal, Ronald.

In: Pediatric Research, Vol. 52, No. 5, 01.11.2002, p. 636-644.

Research output: Contribution to journalArticle

Worgall, Stefan ; Martushova, Katherine ; Busch, Annette ; Lande, Leah ; Crystal, Ronald. / Apoptosis induced by Pseudomonas aeruginosa in antigen presenting cells is diminished by genetic modification with CD40 ligand. In: Pediatric Research. 2002 ; Vol. 52, No. 5. pp. 636-644.
@article{2809976e0f9f4552bef0b5256ad7666b,
title = "Apoptosis induced by Pseudomonas aeruginosa in antigen presenting cells is diminished by genetic modification with CD40 ligand",
abstract = "Persistent colonization with Pseudomonas aeruginosa (PA) is a hallmark of the lung disease associated with cystic fibrosis (CF). Based on the concept that PA is not cleared from the lung by the host response in individuals with CF, we analyzed the capacity of PA to induce cell death in human alveolar macrophages (AM) and murine dendritic cells (DC), antigen presenting cells that play a central role in the initiation of pulmonary host defenses against pathogens, and evaluated if genetic modification can lead to protection against PA induced cell death. AM and DC were susceptible to cell death induced by the laboratory PA isolates PAO1, PAK and PA103, as well as a mucoid derivative of PAO1 and PA isolates derived from sputum of individuals with CF. Apoptosis, analyzed by TUNEL assay, was detectable in AM and DC as early as 3 h after infection with PA. In contrast, the same strains and doses of PA had little effect on the lung epithelial cell line A549 and primary cultures of human bronchial epithelial cells in vitro. Pretreatment of DC with the caspase inhibitors VAD-fmk and YVAD-cmk reduced PA induced cell death (p < 0.05). Finally, genetic modification of DC to express CD40L using an adenovirus vector decreased the susceptibility of DC to cell death induced by PAO1 compared with DC infected with a control Ad vector (p < 0.01). The data demonstrate that DC and AM are susceptible to apoptosis induced by PA and that this response can be partially reversed by genetic modification with CD40L, a CD4+ T cell molecule that plays a central role in activating antigen presenting cells. These observations suggest a potential mechanism contributing to the persistence of PA in CF and suggest that genetic manipulation of antigen presenting cells with anti-apoptotic genes may be able to strengthen host defenses in CF.",
author = "Stefan Worgall and Katherine Martushova and Annette Busch and Leah Lande and Ronald Crystal",
year = "2002",
month = "11",
day = "1",
doi = "10.1203/01.PDR.0000032382.17567.FB",
language = "English",
volume = "52",
pages = "636--644",
journal = "Pediatric Research",
issn = "0031-3998",
publisher = "Lippincott Williams and Wilkins",
number = "5",

}

TY - JOUR

T1 - Apoptosis induced by Pseudomonas aeruginosa in antigen presenting cells is diminished by genetic modification with CD40 ligand

AU - Worgall, Stefan

AU - Martushova, Katherine

AU - Busch, Annette

AU - Lande, Leah

AU - Crystal, Ronald

PY - 2002/11/1

Y1 - 2002/11/1

N2 - Persistent colonization with Pseudomonas aeruginosa (PA) is a hallmark of the lung disease associated with cystic fibrosis (CF). Based on the concept that PA is not cleared from the lung by the host response in individuals with CF, we analyzed the capacity of PA to induce cell death in human alveolar macrophages (AM) and murine dendritic cells (DC), antigen presenting cells that play a central role in the initiation of pulmonary host defenses against pathogens, and evaluated if genetic modification can lead to protection against PA induced cell death. AM and DC were susceptible to cell death induced by the laboratory PA isolates PAO1, PAK and PA103, as well as a mucoid derivative of PAO1 and PA isolates derived from sputum of individuals with CF. Apoptosis, analyzed by TUNEL assay, was detectable in AM and DC as early as 3 h after infection with PA. In contrast, the same strains and doses of PA had little effect on the lung epithelial cell line A549 and primary cultures of human bronchial epithelial cells in vitro. Pretreatment of DC with the caspase inhibitors VAD-fmk and YVAD-cmk reduced PA induced cell death (p < 0.05). Finally, genetic modification of DC to express CD40L using an adenovirus vector decreased the susceptibility of DC to cell death induced by PAO1 compared with DC infected with a control Ad vector (p < 0.01). The data demonstrate that DC and AM are susceptible to apoptosis induced by PA and that this response can be partially reversed by genetic modification with CD40L, a CD4+ T cell molecule that plays a central role in activating antigen presenting cells. These observations suggest a potential mechanism contributing to the persistence of PA in CF and suggest that genetic manipulation of antigen presenting cells with anti-apoptotic genes may be able to strengthen host defenses in CF.

AB - Persistent colonization with Pseudomonas aeruginosa (PA) is a hallmark of the lung disease associated with cystic fibrosis (CF). Based on the concept that PA is not cleared from the lung by the host response in individuals with CF, we analyzed the capacity of PA to induce cell death in human alveolar macrophages (AM) and murine dendritic cells (DC), antigen presenting cells that play a central role in the initiation of pulmonary host defenses against pathogens, and evaluated if genetic modification can lead to protection against PA induced cell death. AM and DC were susceptible to cell death induced by the laboratory PA isolates PAO1, PAK and PA103, as well as a mucoid derivative of PAO1 and PA isolates derived from sputum of individuals with CF. Apoptosis, analyzed by TUNEL assay, was detectable in AM and DC as early as 3 h after infection with PA. In contrast, the same strains and doses of PA had little effect on the lung epithelial cell line A549 and primary cultures of human bronchial epithelial cells in vitro. Pretreatment of DC with the caspase inhibitors VAD-fmk and YVAD-cmk reduced PA induced cell death (p < 0.05). Finally, genetic modification of DC to express CD40L using an adenovirus vector decreased the susceptibility of DC to cell death induced by PAO1 compared with DC infected with a control Ad vector (p < 0.01). The data demonstrate that DC and AM are susceptible to apoptosis induced by PA and that this response can be partially reversed by genetic modification with CD40L, a CD4+ T cell molecule that plays a central role in activating antigen presenting cells. These observations suggest a potential mechanism contributing to the persistence of PA in CF and suggest that genetic manipulation of antigen presenting cells with anti-apoptotic genes may be able to strengthen host defenses in CF.

UR - http://www.scopus.com/inward/record.url?scp=0036840006&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036840006&partnerID=8YFLogxK

U2 - 10.1203/01.PDR.0000032382.17567.FB

DO - 10.1203/01.PDR.0000032382.17567.FB

M3 - Article

C2 - 12409507

AN - SCOPUS:0036840006

VL - 52

SP - 636

EP - 644

JO - Pediatric Research

JF - Pediatric Research

SN - 0031-3998

IS - 5

ER -