Accuri C6 Flow Cytometer System
Originally published online as doi:10.1189/jlb.0209053 on July 28, 2009

Published online before print July 28, 2009
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
jlb.0209053v1
86/5/1159    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Google Scholar
Right arrow Articles by Jann, N. J.
Right arrow Articles by Landmann, R.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jann, N. J.
Right arrow Articles by Landmann, R.
(Journal of Leukocyte Biology. 2009;86:1159-1169.)
© 2009 Society for Leukocyte Biology

Neutrophil antimicrobial defense against Staphylococcus aureus is mediated by phagolysosomal but not extracellular trap-associated cathelicidin

Naja J. Jann*, Mathias Schmaler*, Sascha A. Kristian{dagger},{ddagger}, Katherine A. Radek{ddagger}, Richard L. Gallo{ddagger}, Victor Nizet{dagger}, Andreas Peschel§ and Regine Landmann*,1

* Department of Biomedicine, University Hospital Basel, Basel, Switzerland; Departments of
{dagger} Pediatrics and
{ddagger} Medicine, University of California, San Diego, La Jolla, California, USA; and
§ Department of Medical Microbiology and Hygiene, University of Tübingen, Tübingen, Germany

1. Correspondence: Department of Biomedicine, Division Infection Biology, University Hospital Basel, Hebelstrasse 20, 4031 Basel, Switzerland. E-mail: regine.landmann{at}unibas.ch

ABSTRACT

Neutrophils kill invading pathogens by AMPs, including cathelicidins, ROS, and NETs. The human pathogen Staphylococcus aureus exhibits enhanced resistance to neutrophil AMPs, including the murine cathelicidin CRAMP, in part, as a result of alanylation of teichoic acids by the dlt operon. In this study, we took advantage of the hypersusceptible phenotype of S. aureus {Delta}dltA against cationic AMPs to study the impact of the murine cathelicidin CRAMP on staphylococcal killing and to identify its key site of action in murine neutrophils. We demonstrate that CRAMP remained intracellular during PMN exudation from blood and was secreted upon PMA stimulation. We show first evidence that CRAMP was recruited to phagolysosomes in infected neutrophils and exhibited intracellular activity against S. aureus. Later in infection, neutrophils produced NETs, and immunofluorescence revealed association of CRAMP with S. aureus in NETs, which similarly killed S. aureus wt and {Delta}dltA, indicating that CRAMP activity was reduced when associated with NETs. Indeed, the presence of DNA reduced the antimicrobial activity of CRAMP, and CRAMP localization in response to S. aureus was independent of the NADPH oxidase, whereas killing was partially dependent on a functional NADPH oxidase. Our study indicates that neutrophils use CRAMP in a timed and locally coordinated manner in defense against S. aureus.

Key Words: CRAMP • NETs • peptide • NADPH oxidase • infection