PUBLICATION

The CXCL12/CXCR4 Signaling Axis Retains Neutrophils at Inflammatory Sites in Zebrafish

Authors
Isles, H.M., Herman, K.D., Robertson, A.L., Loynes, C.A., Prince, L.R., Elks, P.M., Renshaw, S.A.
ID
ZDB-PUB-190817-7
Date
2019
Source
Frontiers in immunology   10: 1784 (Journal)
Registered Authors
Elks, Philip, Loynes, Catherine, Renshaw, Steve A., Robertson, Anne
Keywords
CXCL12, CXCR4, chemokine, inflammation, neutrophil, retention, zebrafish
MeSH Terms
  • Animals
  • Neutrophils/immunology*
  • Neutrophils/pathology
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/immunology*
  • Inflammation/genetics
  • Inflammation/immunology
  • Inflammation/pathology
  • Zebrafish/genetics
  • Zebrafish/immunology*
  • Chemokine CXCL12/genetics
  • Chemokine CXCL12/immunology*
  • Cell Movement/genetics
  • Cell Movement/immunology*
  • Gene Knockdown Techniques
  • Receptors, CXCR4/genetics
  • Receptors, CXCR4/immunology*
  • Signal Transduction/genetics
  • Signal Transduction/immunology*
(all 19)
PubMed
31417560 Full text @ Front Immunol
Abstract
The inappropriate retention of neutrophils at inflammatory sites is a major driver of the excessive tissue damage characteristic of respiratory inflammatory diseases including COPD, ARDS, and cystic fibrosis. The molecular programmes which orchestrate neutrophil recruitment to inflammatory sites through chemotactic guidance have been well-studied. However, how neutrophil sensitivity to these cues is modulated during inflammation resolution is not understood. The identification of neutrophil reverse migration as a mechanism of inflammation resolution and the ability to modulate this therapeutically has identified a new target to treat inflammatory disease. Here we investigate the role of the CXCL12/CXCR4 signaling axis in modulating neutrophil retention at inflammatory sites. We used an in vivo tissue injury model to study neutrophilic inflammation using transgenic zebrafish larvae. Expression of cxcl12a and cxcr4b during the tissue damage response was assessed using in situ hybridization and analysis of RNA sequencing data. CRISPR/Cas9 was used to knockdown cxcl12a and cxcr4b in zebrafish larvae. The CXCR4 antagonist AMD3100 was used to block the Cxcl12/Cxcr4 signaling axis pharmacologically. We identified that cxcr4b and cxcl12a are expressed at the wound site in zebrafish larvae during the inflammatory response. Following tail-fin transection, removal of neutrophils from inflammatory sites is significantly increased in cxcr4b and cxcl12a CRISPR knockdown larvae. Pharmacological inhibition of the Cxcl12/Cxcr4 signaling axis accelerated resolution of the neutrophil component of inflammation, an effect caused by an increase in neutrophil reverse migration. The findings of this study suggest that CXCR4/CXCL12 signaling may play an important role in neutrophil retention at inflammatory sites, identifying a potential new target for the therapeutic removal of neutrophils from the lung in chronic inflammatory disease.
Genes / Markers
Figures
Figure Gallery (2 images)
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
i114TgTransgenic Insertion
    s1999tTgTransgenic Insertion
      sh267TgTransgenic Insertion
        w2
          Point Mutation
          1 - 4 of 4
          Show
          Human Disease / Model
          No data available
          Sequence Targeting Reagents
          Target Reagent Reagent Type
          cxcl12aCRISPR1-cxcl12aCRISPR
          cxcr4bCRISPR1-cxcr4bCRISPR
          1 - 2 of 2
          Show
          Fish
          Antibodies
          No data available
          Orthology
          No data available
          Engineered Foreign Genes
          Marker Marker Type Name
          GAL4EFGGAL4
          GFPEFGGFP
          KaedeEFGKaede
          1 - 3 of 3
          Show
          Mapping
          No data available