Subcellular location and properties of bactericidal factors from human neutrophils

doi:10.1084/jem.164.5.1407

This Article

	Full Text (PDF, 841K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JEM
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Gabay, J. E.
	Articles by Nathan, C. F.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Gabay, J. E.
	Articles by Nathan, C. F.


Social Bookmarking

	What's this?

ARTICLES

Subcellular location and properties of bactericidal factors from human neutrophils

JE Gabay, JM Heiple, ZA Cohn and CF Nathan

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 conditions of cell disruption, fractionation, extraction, and assay, almost all BF in human PMN appeared to be localized to the membrane of azurophilic granules as a highly potent, broad-spectrum, rapidly acting protein(s) effective in physiologic medium. Some of these properties appear to distinguish ADBF from previously described PMN bactericidal proteins.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

van Rossum, A. P., Rarok, A. A., Huitema, M. G., Fassina, G., Limburg, P. C., Kallenberg, C. G. M. (2004). Constitutive membrane expression of proteinase 3 (PR3) and neutrophil activation by anti-PR3 antibodies. J. Leukoc. Biol. 76: 1162-1170 [Abstract] [Full Text]
Rarok, A. A., Limburg, P. C., Kallenberg, C. G. M. (2003). Neutrophil-activating potential of antineutrophil cytoplasm autoantibodies. J. Leukoc. Biol. 74: 3-15 [Abstract] [Full Text]
Rarok, A. A., Stegeman, C. A., Limburg, P. C., Kallenberg, C. G. M. (2002). Neutrophil Membrane Expression of Proteinase 3 (PR3) Is Related to Relapse in PR3-ANCA-Associated Vasculitis. J. Am. Soc. Nephrol. 13: 2232-2238 [Abstract] [Full Text]
Parmar, J. S., Mahadeva, R., Reed, B. J., Farahi, N., Cadwallader, K. A., Keogan, M. T., Bilton, D., Chilvers, E. R., Lomas, D. A. (2002). Polymers of alpha 1-Antitrypsin Are Chemotactic for Human Neutrophils . A New Paradigm for the Pathogenesis of Emphysema. Am. J. Respir. Cell Mol. Bio. 26: 723-730 [Abstract] [Full Text]
Ganz, T. (2002). Epithelia: Not just physical barriers. Proc. Natl. Acad. Sci. USA 99: 3357-3358 [Full Text]
Degousee, N., Ghomashchi, F., Stefanski, E., Singer, A., Smart, B. P., Borregaard, N., Reithmeier, R., Lindsay, T. F., Lichtenberger, C., Reinisch, W., Lambeau, G., Arm, J., Tischfield, J., Gelb, M. H., Rubin, B. B. (2002). Groups IV, V, and X Phospholipases A2s in Human Neutrophils. ROLE IN EICOSANOID PRODUCTION AND GRAM-NEGATIVE BACTERIAL PHOSPHOLIPID HYDROLYSIS. J. Biol. Chem. 277: 5061-5073 [Abstract] [Full Text]
Sorensen, O. E., Follin, P., Johnsen, A. H., Calafat, J., Tjabringa, G. S., Hiemstra, P. S., Borregaard, N. (2001). Human cathelicidin, hCAP-18, is processed to the antimicrobial peptide LL-37 by extracellular cleavage with proteinase 3. Blood 97: 3951-3959 [Abstract] [Full Text]
van der Geld, Y. M., Limburg, P. C., Kallenberg, C. G. M. (2001). Proteinase 3, Wegener's autoantigen: from gene to antigen. J. Leukoc. Biol. 69: 177-190 [Abstract] [Full Text]
Risso, A. (2000). Leukocyte antimicrobial peptides: multifunctional effector molecules of innate immunity. J. Leukoc. Biol. 68: 785-792 [Abstract] [Full Text]
Sorensen, O., Arnljots, K., Cowland, J. B., Bainton, D. F., Borregaard, N. (1997). The Human Antibacterial Cathelicidin, hCAP-18, Is Synthesized in Myelocytes and Metamyelocytes and Localized to Specific Granules in Neutrophils. Blood 90: 2796-2803 [Abstract] [Full Text]