Accuri C6 Flow Cytometer System
Originally published online as doi:10.1189/jlb.1108682 on April 23, 2009

Published online before print April 23, 2009
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Buy
Right arrow All Versions of this Article:
jlb.1108682v1
86/1/133    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
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Evans, D. L.
Right arrow Articles by Jaso-Friedmann, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Evans, D. L.
Right arrow Articles by Jaso-Friedmann, L.
(Journal of Leukocyte Biology. 2009;86:133-141.)
© 2009 Society for Leukocyte Biology

Cellular expression and antimicrobial function of a phylogenetically conserved novel histone 1x-like protein on mouse cells: a potential new class of pattern recognition receptor

Donald L. Evans*, Meghan A. Connor*, Lauren D. Moss*, Sarah Lackay{dagger}, John H. Leary, III*, Thomas Krunkosky{ddagger} and Liliana Jaso-Friedmann*,1

* Departments of Infectious Diseases,
{dagger} Pathology, and
{ddagger} Anatomy and Radiology, College of Veterinary Medicine, University of Georgia, Athens, Georgia, USA

1. Correspondence: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA. E-mail: ljaso{at}uga.edu

ABSTRACT

A H1x-like protein (i.e., NCAMP-1) is expressed on the membrane and in GEs from fish NK-like cells. In the present study, we identify the imprinting control region mouse NCAMP-1 ortholog using NCAMP-1 polyclonal antibodies and mAb. Polychromatic flow cytometry revealed NCAMP-1 expression on PBLs (Gr-1+ PMNs were 21.1% NCAMP-1+; DX-5+ NK cells were 12.2% NCAMP-1+), mesenteric LN cells (CD11c+ DCs were 23.2% NCAMP-1+; Gr-1+ PMNs were 24.8% NCAMP-1+; CD21+ B cells were 17.8% NCAMP-1+), and splenocytes (CD11c+ were 39.6% NCAMP-1+; Gr-1+ PMNs were 40.9% NCAMP-1+; DX-5+ NK cells were 24.3% NCAMP-1+; CD21+ B cells were 28.5% NCAMP-1+). Western blot analysis using pNCAMP-1 and GEs from RAW 264.7 cells produced a 32-kDa signal. GEs from RAW 264.7 cells produced a significant reduction in Escherichia coli CFU. This antimicrobial killing activity was inhibited by pretreatment of the extract with (polyclonal) anti-NCAMP-1. Treatment with preimmune serum did not reduce bacterial cell killing. Confocal microscopy using NCAMP-1 and LAMP-1 mAb demonstrated that NCAMP-1 was located on the membrane and in cytosolic vesicles of RAW 264.7 cells and did not appear to colocalize with LAMP-1. NCAMP-1 may participate as a bifunctional protein on cells. It is expressed on the membranes of phagocytic cells, NK cells, and APCs in mice as well as in the granules of macrophages. In phagocytic cells, NCAMP-1 may participate in a nonregulated exocytosis pathway of cellular secretion.

Key Words: innate immune antibacterial peptides • host-pathogen interactions • flow cytometry • macrophages • antigen presenting cells