Toll-like receptor-4 signaling mediates pulmonary neutrophil sequestration in response to gram-positive bacterial enterotoxin

J Surg Res. 2002 May 15;104(2):124-30. doi: 10.1006/jsre.2002.6422.

Abstract

Background: Toll-like receptors (TLRs) serve as mediators of innate immune responses to pathogen-associated molecular patterns (PAMPs) which include lipopolysaccharide (LPS) and staphylococcal enterotoxin B (SEB). TLR-4 is thought to act as the primary effector of LPS recognition and TLR-2 is thought to mediate responses to Gram-positive bacterial proteins. Chemokines such as macrophage inflammatory protein (MIP-2) are peptides that are responsible for lung neutrophil (PMN) sequestration following an infectious or inflammatory insult. Given the Gram-positive origin of SEB, we hypothesized that mice with altered TLR-4 signaling would exhibit no difference in lung PMN sequestration following SEB when compared to wild-type mice.

Methods: Wild-type and TLR-4 mutant mice were administered intratracheal saline, LPS (Escherichia coli 0.1 mg/kg), or SEB (1 mg/kg). After 24 h, lung PMN accumulation was determined by myeloperoxidase (MPO) assay and bronchoalveolar lavage fluid cell count (BALfcc). Total lung and BALf MIP-2 was measured by enzyme-linked immunosorbent assay.

Results: There was an increase in lung PMN accumulation (by both MPO and BALfcc) and MIP-2 following LPS and SEB in wild-type mice compared to saline-treated controls. In contrast, TLR-4 mice failed to exhibit an increase in lung MIP-2 or PMN accumulation following either LPS or SEB compared to wild-type mice.

Conclusions: TLR-4 mutant mice are unresponsive to intratracheal LPS. SEB elicited an increase in lung MIP-2 and PMN accumulation in wild-type mice. However, TLR-4 mutant mice were protected from this process. This suggests that TLR-4 signaling may mediate the responses to other PAMPs in addition to LPS.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Bronchoalveolar Lavage Fluid / chemistry
  • Bronchoalveolar Lavage Fluid / cytology
  • Chemokine CXCL2
  • Drosophila Proteins*
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli
  • Lipopolysaccharides / administration & dosage
  • Lipopolysaccharides / pharmacology*
  • Lung / metabolism
  • Lung / pathology*
  • Male
  • Membrane Glycoproteins / physiology*
  • Mice
  • Mice, Inbred C3H
  • Monokines / analysis
  • Monokines / metabolism
  • Neutrophils / pathology*
  • Peroxidase / analysis
  • Pulmonary Alveoli / pathology
  • Receptors, Cell Surface / physiology*
  • Signal Transduction*
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Toll-Like Receptors
  • Trachea

Substances

  • Chemokine CXCL2
  • Drosophila Proteins
  • Lipopolysaccharides
  • Membrane Glycoproteins
  • Monokines
  • Receptors, Cell Surface
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Toll-Like Receptors
  • Peroxidase