Interleukin (IL)-8 (CXCL8) induces cytokine expression and superoxide formation by guinea pig neutrophils infected with Mycobacterium tuberculosis

Mark J. Lyons, Teizo Yoshimura, David N. McMurray

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Setting: Interleukin (IL)-8, a neutrophil attracting chemokine, is known to be made by a variety of leukocyte populations following stimulation by Mycobacterium tuberculosis. Objective: The effect of recombinant guinea pig IL-8 on the ability of neutrophils to generate a cytokine response after infection with M. tuberculosis H37Ra was examined. Design: Recombinant gpIL-8 was produced by subcloning the gene into Escherichia coli and purification over a nickel column. The identity of the rgpIL-8 was confirmed by sequencing. Neutrophils were harvested from the blood of non-vaccinated or M. bovis BCG-vaccinated guinea pigs and tested for their ability to migrate toward media alone, 10μg/ml PPD, f-Met-Leu-Phe (f-MLP), or rgpIL-8 in 96-well chemotactic chambers. Neutrophils were also pre-stimulated with rgpIL-8 then restimulated with LPS (10μg/ml) or infected in vitro with M. tuberculosis H37Ra (MOI 1:1). Results: Recombinant gpIL-8 and f-MLP induced significant chemotaxis in neutrophils from both non-vaccinated and BCG-vaccinated guinea pigs, with the best chemotaxis occuring at a concentration of 10-7M. Real-time PCR analysis revealed that pre-treatment of neutrophils induced elevated levels of IL-8 and TNF-α mRNA and protein as well as superoxide, but not mRNA for MCP-1, IFN-γ, or TGF-β when compared to neutrophils pre-stimulated with media alone. Conclusions: The presence of IL-8 early in the host response to M. tuberculosis infection may be an important contributor to a successful immune response. How essential a role IL-8 plays remains unknown and merits further study.

Original languageEnglish
Pages (from-to)283-292
Number of pages10
JournalTuberculosis
Volume84
Issue number5
DOIs
Publication statusPublished - 2004
Externally publishedYes

Fingerprint

interleukin-8
Mycobacterium tuberculosis
Interleukin-8
Superoxides
superoxide anion
guinea pigs
neutrophils
Guinea Pigs
Neutrophils
cytokines
Cytokines
methionyl-leucyl-phenylalanine
Aptitude
chemotaxis
Chemotaxis
Mycobacterium bovis
Messenger RNA
Mycobacterium Infections
Tuberculin
1-methylcyclopropene

Keywords

  • Guinea pig
  • IL-8
  • Neutrophils

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Microbiology
  • Immunology and Allergy
  • Infectious Diseases
  • veterinary(all)

Cite this

Interleukin (IL)-8 (CXCL8) induces cytokine expression and superoxide formation by guinea pig neutrophils infected with Mycobacterium tuberculosis. / Lyons, Mark J.; Yoshimura, Teizo; McMurray, David N.

In: Tuberculosis, Vol. 84, No. 5, 2004, p. 283-292.

Research output: Contribution to journalArticle

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abstract = "Setting: Interleukin (IL)-8, a neutrophil attracting chemokine, is known to be made by a variety of leukocyte populations following stimulation by Mycobacterium tuberculosis. Objective: The effect of recombinant guinea pig IL-8 on the ability of neutrophils to generate a cytokine response after infection with M. tuberculosis H37Ra was examined. Design: Recombinant gpIL-8 was produced by subcloning the gene into Escherichia coli and purification over a nickel column. The identity of the rgpIL-8 was confirmed by sequencing. Neutrophils were harvested from the blood of non-vaccinated or M. bovis BCG-vaccinated guinea pigs and tested for their ability to migrate toward media alone, 10μg/ml PPD, f-Met-Leu-Phe (f-MLP), or rgpIL-8 in 96-well chemotactic chambers. Neutrophils were also pre-stimulated with rgpIL-8 then restimulated with LPS (10μg/ml) or infected in vitro with M. tuberculosis H37Ra (MOI 1:1). Results: Recombinant gpIL-8 and f-MLP induced significant chemotaxis in neutrophils from both non-vaccinated and BCG-vaccinated guinea pigs, with the best chemotaxis occuring at a concentration of 10-7M. Real-time PCR analysis revealed that pre-treatment of neutrophils induced elevated levels of IL-8 and TNF-α mRNA and protein as well as superoxide, but not mRNA for MCP-1, IFN-γ, or TGF-β when compared to neutrophils pre-stimulated with media alone. Conclusions: The presence of IL-8 early in the host response to M. tuberculosis infection may be an important contributor to a successful immune response. How essential a role IL-8 plays remains unknown and merits further study.",
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N2 - Setting: Interleukin (IL)-8, a neutrophil attracting chemokine, is known to be made by a variety of leukocyte populations following stimulation by Mycobacterium tuberculosis. Objective: The effect of recombinant guinea pig IL-8 on the ability of neutrophils to generate a cytokine response after infection with M. tuberculosis H37Ra was examined. Design: Recombinant gpIL-8 was produced by subcloning the gene into Escherichia coli and purification over a nickel column. The identity of the rgpIL-8 was confirmed by sequencing. Neutrophils were harvested from the blood of non-vaccinated or M. bovis BCG-vaccinated guinea pigs and tested for their ability to migrate toward media alone, 10μg/ml PPD, f-Met-Leu-Phe (f-MLP), or rgpIL-8 in 96-well chemotactic chambers. Neutrophils were also pre-stimulated with rgpIL-8 then restimulated with LPS (10μg/ml) or infected in vitro with M. tuberculosis H37Ra (MOI 1:1). Results: Recombinant gpIL-8 and f-MLP induced significant chemotaxis in neutrophils from both non-vaccinated and BCG-vaccinated guinea pigs, with the best chemotaxis occuring at a concentration of 10-7M. Real-time PCR analysis revealed that pre-treatment of neutrophils induced elevated levels of IL-8 and TNF-α mRNA and protein as well as superoxide, but not mRNA for MCP-1, IFN-γ, or TGF-β when compared to neutrophils pre-stimulated with media alone. Conclusions: The presence of IL-8 early in the host response to M. tuberculosis infection may be an important contributor to a successful immune response. How essential a role IL-8 plays remains unknown and merits further study.

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