Separation of sublethal and lethal effects of polymorphonuclear leukocytes on Escherichia coli.

B A Mannion; J Weiss; P Elsbach

doi:10.1172/JCI114755

Separation of sublethal and lethal effects of polymorphonuclear leukocytes on Escherichia coli.

B A Mannion, J Weiss, and P Elsbach

Published August 1, 1990 - More info

View PDF

Abstract

Escherichia coli ingested by PMN promptly stop growing and form no colonies in nutrient agar, but metabolize near normally for up to several hours. The bactericidal/permeability increasing protein (BPI) of PMN also inhibits E. coli growth without initial metabolic impairment. We recently showed that BPI-treated E. coli, although unable to grow in normal nutrient agar, can form colonies in this medium plus 0.1% BSA, as long as their metabolism is maintained, indicating that biochemical impairment is a better indicator of death than growth arrest (1990. J. Clin. Invest. 85:853-860). We have now reexamined the fate of ingested E. coli. Rabbit PMN ingest greater than 85% of several rough E. coli strains in 15 min, but greater than 80% of these bacteria, while unable to form colonies in conventional agar, grow normally on agar plus 0.1% BSA. Thus, the PMN under these conditions promptly stop growth of ingested E. coli without killing. Adding nonlethal concentrations of normal human serum (NHS) before, but not after ingestion, accelerates killing and, in parallel, loss of bacterial metabolism (t1/2 less than 0.5 h vs. greater than 3 h, respectively, with and without NHS). The rapid killing of both rough and smooth E. coli pretreated with NHS is lost after C7 depletion (C7-D) and restored when C7 is replenished. Similar results are obtained with human PMN. In contrast, ingested Staphylococcus epidermidis, opsonized with either NHS or C7-D serum rapidly stop metabolizing and do not form colonies in nutrient agar with or without BSA. Respiratory burst activity is the same during ingestion of E. coli (with or without NHS) and S. epidermidis. Killing of E. coli J5 (however, not of O111-B4) by BPI is also accelerated by pretreatment with NHS but not C7-D human serum. These findings indicate that late complement components are needed for efficient killing of both rough and smooth E. coli by PMN, and that BPI is the principal intracellular agent acting on ingested rough E. coli.

Images.