Monitoring of neutrophil priming in whole blood by antibodies isolated from a synthetic phage antibody library

J Leukoc Biol. 2000 Jul;68(1):58-64.


Neutrophil activation is a multistep process. In vitro activation of neutrophils with semiphysiological activators is optimal only after preactivation or priming with cytokines, chemotaxins, and/or bacterial products. Until now, no antibodies have been developed that can distinguish between resting and (cytokine) primed neutrophils with a sufficient dynamic range necessary for screening clinical samples. We have isolated two human phage antibodies, designated MoPhab A17 and A27, from a synthetic bacteriophage antibody library. These phage antibodies recognize epitopes that are upregulated on neutrophils present in whole blood treated with low priming concentrations of cytokines, such as GM-CSF and TNF-alpha. This induction was time- and concentration-dependent and optimal at concentrations that are sufficient for priming functional responses in neutrophils: GM-CSF (10 pM) and TNF-alpha (100 IU/ml). PMNs, isolated from the peripheral blood of chronic obstructive pulmonary disease (COPD) patients with a clinical exacerbation, exhibited a partial in vivo primed phenotype. These antibodies promise to be an ideal tool to monitor disease activity in whole blood of patients with inflammatory diseases.

MeSH terms

  • Adult
  • Antibodies, Monoclonal / pharmacology*
  • Bacteriophages / genetics
  • Epitopes / immunology
  • Genes, Immunoglobulin
  • Genetic Vectors
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology*
  • Humans
  • Immunoglobulin Fragments / genetics
  • Immunoglobulin Fragments / pharmacology*
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Variable Region / genetics
  • Lung Diseases, Obstructive / blood
  • Lung Diseases, Obstructive / immunology
  • Lung Diseases, Obstructive / pathology
  • Neutrophils / drug effects
  • Neutrophils / physiology*
  • Peptide Fragments / chemical synthesis
  • Peptide Fragments / genetics
  • Peptide Fragments / pharmacology*
  • Phagocytosis / drug effects
  • Tumor Necrosis Factor-alpha / pharmacology*


  • Antibodies, Monoclonal
  • Epitopes
  • Immunoglobulin Fragments
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • Peptide Fragments
  • Tumor Necrosis Factor-alpha
  • Granulocyte-Macrophage Colony-Stimulating Factor