Isolation, characterization, cDNA cloning, and antimicrobial properties oftwo distinct subfamilies of alpha-defensins from rhesus macaque leukocytes

Experiments to isolate and characterize rhesus macaque myeloid alpha-defens ins (RMADs) were conducted, Seven RMAD peptides were isolated and sequenced , and the cDNAs encoding six of these peptides and one other alpha-defensin from bone marrow were also characterized. Four of the RMADs were found to be highly similar to human neutrophil alpha-defensins HNP-1 to HNP-3, while the remaining four peptides were much more similar to human enteric alpha- defensin HD-5. Two alpha-defensin pairs differed only by the presence or ab sence of an additional arginine at the amino termini of their mature peptid es, indicative of alternate posttranslational processing, The primary trans lation products of RMAD-1 to -8 are 94- and 96-amino-acid prepropeptides th at are highly similar to those of human alpha-defensins. Immunolocalization experiments revealed a granular cytoplasmic pattern in the cytoplasms of n eutrophils, indistinguishable from the pattern observed after immunostainin g of human myeloid alpha-defensins in polymorphonuclear leukocytes, Each of the purified peptides was tested for its in vitro activities against Staph ylococcus aureus 502a, Listeria monocytogenes EGD, Escherichia coil ML35, a nd Cryptococcus neoformans 271A. Several of the peptides were microbicidal for the gram-positive bacteria and C. neoformans at defensin concentrations in the range of 2 to 5 mu M All of the peptides were bacteriostatic agains t E. call, but none were bactericidal for this organism. This study is the first to characterize the sequences and activities of alpha-defensins from nonhuman primates, data that should aid in delineating the role of these pe ptides in rhesus macaque host defense.