Measurement of bacterial ingestion and killing by macrophages

Current Protocols in Immunology
P A CampbellD A Drevets

Abstract

This unit presents fairly simple assays for measuring the binding of bacteria to macrophages, internalization of bacteria (also called ingestion or phagocytosis), and bacterial killing by macrophages. The first basic protocol describes how to measure the ability of macrophages to ingest bacteria. Because it is critical to remove residual extracellular organisms, the protocol presents two alternative steps to accomplish this: a washing procedure and a more stringent method in which cells are sedimented through sucrose. In addition, it is important to distinguish those bacteria truly ingested by a macrophage from those that are bound to, but not internalized by, the cell. A simple but effective way to do this is described in an alternate protocol. The unit also presents two ways to measure the ability of a macrophage to kill bacteria it has internalized. The first is a straightforward assay in which bacterial colonies are enumerated before and after a killing period; a subsequent colony count will indicate whether the bacteria grew within or were killed by the macrophage. The second protocol describes a way to measure bacterial viability based on bacterial metabolism, in which the ability of bacterial dehydrogenases to mediate th...Continue Reading

References

Nov 1, 1987·The Journal of Experimental Medicine·N R Payne, M A Horwitz
Sep 3, 1985·Journal of Immunological Methods·R Peck
Jun 1, 1994·Infection and Immunity·D A DrevetsP A Campbell

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Citations

Aug 13, 2013·PLoS Pathogens·Christopher L NoblesAnthony W Maresso
Sep 21, 2004·American Journal of Physiology. Lung Cellular and Molecular Physiology·Anatoly N MikerovJoanna Floros
Nov 6, 2007·American Journal of Physiology. Lung Cellular and Molecular Physiology·Anatoly N MikerovJoanna Floros
Dec 19, 2007·European Journal of Immunology·Tobias FrankenbergGeorg Häcker
Nov 16, 2011·Infection and Immunity·Brandon M ShawKevin Dybvig
Jan 9, 2010·Microbiology and Immunology·Cíntia Silva dos SantosAna Luíza de Mattos-Guaraldi
Sep 10, 1999·Microbiology and Immunology·M HeinzelmannH C Polk
Apr 22, 2015·The Journal of Immunology : Official Journal of the American Association of Immunologists·S Marina Casalino-MatsudaPeter H S Sporn
Feb 5, 2016·Analytical and Bioanalytical Chemistry·Guat Wei LimDerek Juinn Chieh Chan
Oct 3, 2015·In Vitro Cellular & Developmental Biology. Animal·Wenhui QiuMing Yang
Sep 16, 2015·The Journal of Immunology : Official Journal of the American Association of Immunologists·Evan L ChiswickDaniel G Remick
Jan 15, 2003·Journal of Virology·Kazuhiro KondoKoichi Yamanishi
Dec 21, 2017·Environmental Science & Technology·Wenhui QiuKe-Jian Wang
Dec 22, 2007·Nanomedicine·Jennifer B HallScott E McNeil

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