Use of UV-irradiated bacteriophage T6 to kill extracellular bacteria in tissue culture infectivity assays

Denise R. Shaw, Anthony T. Maurelli, Jon D. Goguen, Susan C. Straley, Roy Curtiss

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

We have utilized 'lysis from without' mediated by UV-inactivated bacteriophage T6 to eliminate extracellular bacteria in experiments measuring the internalization, intracellular survival and replication of Yersinia pestis within mouse peritoneal macrophages and of Shigella flexneri within a human intestinal epithelial cell line. The technique we describe has the following characteristics: (a) bacterial killing is complete within 15 min at 37°C, with a > 103-fold reduction in colony-forming units (CFU); (b) bacteria within cultured mammalian cells are protected from killing by UV-inactivated T6; (c) the mammalian cells are not observably affected by exposure to UV-inactivated T6. This technique has several advantages over the use of antibiotics to eliminate extracellular bacteria and is potentially widely applicable in studies of the interactions between pathogenic bacteria and host phagocytic cells as well as other target tissues.

Original languageEnglish (US)
Pages (from-to)75-83
Number of pages9
JournalJournal of Immunological Methods
Volume56
Issue number1
DOIs
StatePublished - Jan 14 1983
Externally publishedYes

Keywords

  • bacteria-phagocyte interactions
  • bactericidal assays
  • in vitro infectivity assays
  • intracellular bacterial pathogens
  • phagocytosis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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