A recombinant 63-kDa form of Bacillus anthracis protective antigen produced in the yeast Saccharomyces cerevisiae provides protection in rabbit and primate inhalational challenge models of anthrax infection.
Infection by Bacillus anthracis is preventable by prophylactic vaccination with several naturally derived and recombinant vaccine preparations. Existing data suggests that protection is mediated by antibodies directed against the protective antigen (PA) component of the anthrax toxin complex. PA is an 83-kDa protein cleaved in vivo to yield a biologically active 63-kDa protein. In an effort to evaluate the potential of yeast as an expression system for the production of recombinant PA, and to determine if the yeast-purified rPA63 can protect from a lethal inhalational challenge, the sequence of the 63-kDa form of PA was codon-optimized and expressed in the yeast Saccharomyces cerevisiae. Highly purified rPA63 isolated from Saccharomyces under denaturing conditions demonstrated reduced biological activity in a macrophage-killing assay compared to non-denatured rPA83 purified from Escherichia coli. Rabbits and non-human primates (NHP) immunized with rPA63 and later challenged with a lethal dose of B. anthracis spores were generally protected from infection. These results indicate that epitopes present in the 63-kDa from of PA can protect rabbits and non-human primates from a lethal spore challenge, and further suggest that a fully functional rPA63 is not required in order to provide these epitopes.
Hepler
Robert W
RW
Merck Research Laboratories, West Point, PA 19486, USA.
Kelly
Rosemarie
R
McNeely
Tessie B
TB
Fan
Hongxia
H
Losada
Maria C
MC
George
Hugh A
HA
Woods
Andrea
A
Cope
Leslie D
LD
Bansal
Alka
A
Cook
James C
JC
Zang
Gina
G
Cohen
Steven L
SL
Wei
Xiaorong
X
Keller
Paul M
PM
Leffel
Elizabeth
E
Joyce
Joseph G
JG
Pitt
Louise
L
Schultz
Loren D
LD
Jansen
Kathrin U
KU
Kurtz
Myra
M
D000595:Amino Acid Sequence; D000818:Animals; D000881:Anthrax / Q000517:prevention & control*; D022122:Anthrax Vaccines / Q000276:immunology*; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D003062:Codon; D005260:Female; D008253:Macaca mulatta; D008297:Male; D008969:Molecular Sequence Data; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014614:Vaccines, Synthetic / Q000276:immunology*
D022122:Anthrax Vaccines; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D003062:Codon; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; C030325:anthrax toxin