Respiratory burst facilitates the digestion of Escherichia coli killed by polymorphonuclear leukocytes.
We examined factors that may limit degradation of bacterial protein of Escherichia coli S15 killed by polymorphonuclear leukocytes (PMN). Both human and rabbit PMN degraded up to 40% of [14C]amino acid-labeled protein of ingested and killed E. coli in 2 h as determined by loss of acid-precipitable radioactivity. In contrast, equally bactericidal broken-PMN preparations or isolated granules degraded only about 10% of bacterial protein regardless of pH. To determine whether activation of the respiratory burst contributes to digestion, we compared degradation by intact PMN in room air and under N2. Depletion of O2 by N2 flushing had no effect on the bactericidal activity of either human or rabbit PMN but reduced degradation by approximately 50%. Protein degradation during phagocytosis was also reduced in the presence of cyanide or azide, inhibitors of myeloperoxidase (MPO). PMN of two patients with chronic granulomatous disease ingested and killed E. coli S15 as well as did normal PMN but degraded bacterial protein as did normal PMN incubated under N2. The low degradative activity of PMN disrupted by sonication could be raised to nearly the level of intact PMN incubated in room air by preincubation of the PMN with 10(-7) M formyl-methionyl-leucyl-phenylalanine (fMLP) before sonication and by pretreatment of E. coli with MPO. Depletion of O2 or chloride during these preincubations with formyl-methionyl-leucyl-phenylalanine respectively, virtually abolished and markedly diminished stimulation of bacterial protein degradation. We conclude that enhanced MPO-mediated O2 metabolism of intact PMN plays a role in the digestion of killed E. coli.
Weiss
J
J
Kao
L
L
Victor
M
M
Elsbach
P
P
D000818:Animals; D001386:Azides / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism; D001770:Blood Bactericidal Activity / Q000187:drug effects; D002474:Cell-Free System; D003486:Cyanides / Q000494:pharmacology; D004926:Escherichia coli / Q000276:immunology* / Q000378:metabolism; D006105:Granulomatous Disease, Chronic / Q000503:physiopathology; D006801:Humans; D009504:Neutrophils / Q000502:physiology*; D010100:Oxygen / Q000378:metabolism; D010101:Oxygen Consumption; D011817:Rabbits
D001386:Azides; D001426:Bacterial Proteins; D003486:Cyanides; D010100:Oxygen