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(Journal of Leukocyte Biology. 2000;68:58-64.)
© 2000 by Society for Leukocyte Biology

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

Leo Koenderman, Deon Kanters, B. Maesen, Jan Raaijmakers, Jan-Willem J. Lammers, John de Kruif*,{dagger} and Ton Logtenberg*,{dagger}

Department of Pulmonary Diseases and
* Immunology, University Medical Center and
{dagger} Utrecht Biotechnology Systems, Utrecht, The Netherlands

Correspondence: Dr. L. Koenderman, Department of Pulmonary Diseases, F.02.333, University Hospital Utrecht, Heidelberglaan 100, NL 3508 GA Utrecht, The Netherlands. E-mail: L.Koenderman{at}hli.azu.nl

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.

Key Words: priming • detection • antibodies • neutrophils • monocytes




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