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

Proc Natl Acad Sci U S A. 1981 Dec;78(12):7540-4. doi: 10.1073/pnas.78.12.7540.

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.

Publication types

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

MeSH terms

  • Antigen-Antibody Complex
  • Chemotactic Factors / metabolism*
  • Chemotaxis, Leukocyte*
  • Fluoresceins
  • Humans
  • Immunologic Techniques
  • Ligands
  • Neutrophils / physiology*
  • Oligopeptides / metabolism*
  • Time Factors

Substances

  • Antigen-Antibody Complex
  • Chemotactic Factors
  • Fluoresceins
  • Ligands
  • Oligopeptides
  • F-chemotactic peptide