Interleukin 2 promoter/enhancer controlled expression of a synthetic cecropin-class lytic peptide in transgenic mice and subsequent resistance to Brucella abortus.
The addition of an antimicrobial that can be synthesized by the mammalian immune system at the point of challenge may enhance disease resistance. A possible group of agents are cecropins, broad-spectrum antimicrobial peptides, which have been described and characterized. They are relatively non-toxic to normal cells from multicellular organisms but are toxic to a wide range of bacteria, protozoa and fungi, as well as infected and abnormal cells. Twenty-six lines of transgenic mice were produced by pronuclear injection of DNA consisting of the 5'-flanking region from -593 to +110 of the mouse interleukin 2 (IL-2) gene, Shiva 1a (a synthetic cecropinclass lytic peptide), and the SV40 polyadenylation/splice signal. A reverse-transcription PCR assay determined that two lines of transgenic mice were produced whose spleen-derived lymphocytes could be induced to transcribe and mature mRNA for Shiva 1a by exposure to 3.25 mg ml-1 of Con A. Two lines were challenged with an inoculation of 5 x 10(4) Brucella abortus strain 2308. After four weeks, there were significantly fewer B. abortus organisms in the spleens of transgenic mice than in non-transgenic control mice of the same strain (p < 0.05). Since the controlling regions of the IL-2 enhancer and the amino acid sequence of the signal peptide are highly conserved among several species, it is likely that this recombinant gene will function in other mammals.
Reed
W A
WA
Department of Animal, Dairy, and Veterinary Sciences, Logan, Utah State University 84322-4700, USA.
Elzer
P H
PH
Enright
F M
FM
Jaynes
J M
JM
Morrey
J D
JD
White
K L
KL
D000818:Animals; D023181:Antimicrobial Cationic Peptides; D002003:Brucella abortus; D002006:Brucellosis / Q000276:immunology / Q000517:prevention & control; D004742:Enhancer Elements, Genetic; D004926:Escherichia coli / Q000187:drug effects; D005260:Female; D007376:Interleukin-2 / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic / Q000235:genetics*; D010455:Peptides / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013154:Spleen / Q000382:microbiology; D019076:Transgenes
D023181:Antimicrobial Cationic Peptides; D007376:Interleukin-2; D010455:Peptides; D011993:Recombinant Fusion Proteins; C057355:Shiva-1