Synthetic peptide chemotactic factors for neutrophils: the range of active peptides, their efficacy and inhibitory activity, and susceptibility of the cellular response to enzymes and bacterial toxins

Abstract

The chemotactic activity for neutrophil leucocytes of twenty-six peptides of varied sequence, of which the majority were N-formylated, was assessed by determining the concentration at which each was maximally active and the efficacy of each peptide at that concentration. These two measures of activity did not correlate with one another. Many formylated peptides with a wide variety of sequences were active. Of these, the formyl-methionyl peptides had highest efficacy, but many other peptides were active at concentrations as low as the formyl-methionyl tripeptides. Unrelated peptides, viz formyl-methionyl-leucyl-phenylalanine, acetyl-tri-alanine, formyl-tri--phenyla-lanine, cross-inhibit the cells' response to one another, and this inhibition is reversible. Inhibition is prevented if the cells are incubated throughout the experiment in levamisole or A23187. These experiments suggest that the leucocyte peptide receptor is capable of binding many ligands, and that activation of a response is not solely a function of binding affinity. They exclude a strict steric specificity for binding. Chemotactic responses to formylated peptides were shown to be reduced in cells pretreated with perfringolysin, a bacterial cholesterol-binding toxin, and with phospholipase C. Trypsin and pronase also reduced these responses when used at 500 micrograms per 10(6) cells but not at lower doses.