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Infection and Immunity, March 2004, p. 1470-1478, Vol. 72, No. 3
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.3.1470-1478.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Paneth Cell {alpha}-Defensins from Rhesus Macaque Small Intestine

Hiroki Tanabe,1,{dagger} Jun Yuan,1 Melinda M. Zaragoza,2 Satya Dandekar,2,3 Agnes Henschen-Edman,4 Michael E. Selsted,1,5 and Andre J. Ouellette1,5*

Departments of Pathology,1 Microbiology and Molecular Genetics, College of Medicine,5 Department of Molecular Biology and Biochemistry, School of Biological Sciences, University of California, Irvine, California 92697,4 Department of Microbiology and Immunology, School of Medicine,2 California National Primate Research Center, University of California, Davis, California 956163

Received 22 October 2003/ Returned for modification 6 November 2003/ Accepted 17 November 2003

Antimicrobial peptides are secreted by small intestinal Paneth cells as components of innate immunity. To investigate the role of {alpha}-defensins in enteric host defenses in nonhuman primates, {alpha}-defensin cDNAs were isolated, {alpha}-defensin peptides were purified from rhesus macaque small bowel, and the bactericidal activities of the peptides were measured. Six rhesus enteric {alpha}-defensin (RED) cDNAs, RED-1 to RED-6, were identified in a jejunum cDNA library; the deduced RED peptides exhibited extensive diversity relative to the primary structures of rhesus myeloid {alpha}-defensins. RED-4 was purified from monkey jejunum, and N-terminal peptide sequencing of putative RED-4 peptides identified two N termini, RTCYCRTGR... and TCYCRTGRC...; these corresponded to alternative N termini for the RED-4 molecules, as deduced from their molecular masses and RED cDNAs. In situ hybridization experiments localized RED mRNAs exclusively to small intestinal Paneth cells. Recombinant RED-1 to RED-4 were purified to homogeneity and shown to be microbicidal in the low micromolar range (<=10 µg/ml) against gram-positive and gram-negative bacteria, with individual peptides exhibiting variable target cell specificities. Thus, compared to myeloid {alpha}-defensins from rhesus macaques, enteric {alpha}-defensin peptides are highly variable in both primary structure and activity. These studies should facilitate further analyses of the role of {alpha}-defensins in primate enteric immunity.

* Corresponding author. Mailing address: Department of Pathology, Med Sci I D-440, College of Medicine, University of California, Irvine, CA 92697-4800. Phone: 949-824-4647. Fax: 949-824-1098. E-mail: aouellet{at}uci.edu.

Editor: F. C. Fang

{dagger} Present address: Sano Hospital, 3-3-1-15 Suehiro, Asahikawa, Hokkaido 071-8133, Japan.

Infection and Immunity, March 2004, p. 1470-1478, Vol. 72, No. 3
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.3.1470-1478.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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