PMID: 3772295Nov 1, 1986Paper

Subcellular location and properties of bactericidal factors from human neutrophils

The Journal of Experimental Medicine
J E GabayC F Nathan

Abstract

We examined the subcellular location of bactericidal factors (BF) in human neutrophils, using an efficient fractionation scheme. Nitrogen bomb cavitates of DIFP-treated PMN were centrifuged through discontinuous Percoll gradients, each fraction extracted with 0.05 M glycine, pH 2.0, and tested for the killing of Escherichia coli. greater than 90% of BF coisolated with the azurophil granules. After lysis of azurophils, 98% of azurophil-derived BF (ADBF) sedimented with the membrane. ADBF activity was solubilized from azurophil membrane with either acid or nonionic detergent (Triton X-100, Triton X-114). Bactericidal activity was linear with respect to protein concentration over the range 0.3-30 micrograms/ml. 0.1-0.3 microgram/ml ADBF killed 10(5) E. coli within 30 min at 37 degrees C. At 1.4 micrograms/ml, 50% of 2 X 10(5) bacteria were killed within 5 min. ADBF was effective between pH 5-8, with peak activity at pH 5.5. Glucose (20 mM), EDTA (1-25 mM), and physiologic concentrations of NaCl or KCl had little or no inhibitory effect on ADBF. ADBF killed both Gram-positive and Gram-negative virulent clinical isolates, including listeria, staphylococci, beta-hemolytic streptococci, and Pseudomonas aeruginosa. Thus, under these co...Continue Reading

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Citations

Jul 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·J E GabayC F Nathan
Dec 1, 1988·The Journal of Experimental Medicine·M McGroganJ Gabay
Jan 1, 1994·The Journal of Experimental Medicine·R M Steinman, C L Moberg
May 20, 2004·Kidney International·Adrian SchreiberRalph Kettritz
Jan 1, 1994·Mediators of Inflammation·V Witko-Sarsat, B Descamps-Latscha
May 30, 2002·American Journal of Respiratory Cell and Molecular Biology·Jasvir S ParmarDavid A Lomas
Apr 1, 1988·The Journal of Clinical Investigation·D L GrangerD T Durack
Mar 21, 2002·Proceedings of the National Academy of Sciences of the United States of America·Tomas Ganz
Oct 29, 2005·Scandinavian Journal of Immunology·J O GrönroosV J O Laine
Aug 5, 2008·Biochemical and Biophysical Research Communications·Shintaro HaraShigeru Kohno
Aug 28, 2004·Journal of Leukocyte Biology·André P van RossumCees G M Kallenberg
Jul 2, 2003·Journal of Leukocyte Biology·Agnieszka A RarokCees G M Kallenberg
Jan 1, 1990·European Journal of Haematology·T GanzR I Lehrer
Aug 23, 2002·Journal of the American Society of Nephrology : JASN·Agnieszka A RarokCees G M Kallenberg
Jul 1, 1995·Clinical Microbiology Reviews·D K Wagner, P G Sohnle

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