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Fluoresceinated chemotactic peptide and high-affinity anti-fluorescein antibody as a probe of the temporal characteristics of neutrophil stimulation

Academic Article
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Overview

authors

  • Sklar, L. A.
  • Oades, Z. G.
  • Jesaitis, A. J.
  • Painter, R. G.
  • Cochrane, Charles

publication date

  • 1981

journal

  • Proceedings of the National Academy of Sciences of the United States of America-Biological Sciences  Journal

abstract

  • Antifluorescein antibody molecules were used to interrupt the stimulation of neutrophils by a fluoresceinated chemotactic peptide. From the results we construct a semiquantitative relationship among ligand-receptor interaction, the time course of cell triggering and response, and aspects of cellular adaptation. The interaction of the antibody with the free fluoresceinated peptide is complete within a few seconds and the peptide-antibody complex neither stimulates the cells nor inhibits subsequent stimulation by unlabeled peptide. When antibody is added to a cell suspension that has been stimulated with the fluoresceinated peptide, we observe that: (i) the apparent membrane depolarization response monitored by a fluorescent dye can be inhibited only if antibody is added within 30 sec of stimulation; (ii) the superoxide response can be inhibited even if antibody is added more than 1 min after stimulation and decays with an intrinsic half-life of about 12 sec; (iii) responses to a second dose of nonfluoresceinated peptide are enhanced if the antibody is added within 2 min of stimulation by the fluoresceinated peptide. These results suggest that different neutrophil responses depend in individual ways on the time course and extent of ligand binding to its receptor. A comparison of these data with the time course of binding permits an estimate of the number of receptors involved in these responses.

subject areas

  • Antigen-Antibody Complex
  • Chemotactic Factors
  • Chemotaxis, Leukocyte
  • Fluoresceins
  • Humans
  • Immunologic Techniques
  • Ligands
  • Neutrophils
  • Oligopeptides
  • Time Factors
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Identity

PubMed Central ID

  • PMC349304

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.78.12.7540

PubMed ID

  • 7038681
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Additional Document Info

start page

  • 7540

end page

  • 7544

volume

  • 78

issue

  • 12

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