Identification of in vivo-induced conserved sequences from Yersinia pestis during experimental plague infection in the rabbit.
In an effort to identify the novel virulence determinants of Yersinia pestis, we applied the gene "discovery" methodology, in vivo-induced (IVI) antigen technology, to detect genes upregulated during infection in a laboratory rabbit model for bubonic plague. After screening over 70,000 Escherichia coli clones of Y. pestis DNA expression libraries, products from 25 loci were identified as being seroreactive to reductively adsorbed, pooled immune serum. Upon sequence analysis of the predicted IVI gene products, more frequently encountered conserved protein functional categories have emerged, to include type-V autotransporters and components of more complex secretion systems including types III and VI. The recombinant products from eight selected clones were subsequently immunoblotted against pooled immune serum from two naturally infected host species: the prairie dog, and a species refractory to lethal disease, the coyote. Immune prairie dog serum recognized 2-3 of the rabbit-reactive antigens, suggesting at least some overlap in the pathogen's in vivo survival mechanisms between these two hosts. Although the coyote serum failed to recognize most of the IVI antigens, LepA was universally reactive with all three host sera. Collectively, the profiles/patterns of IVI conserved sequences (IVICS) may represent immune "signatures" among different host species, possessing the potential for use as a diagnostic tool for plague. Further, the antigenic nature of IVICS makes them ideal for further evaluation as novel subunit vaccine candidates. The gathering of additional data and analysis of the intact IVI genes and the expressed IVICS products should provide insight into the unique biologic processes of Y. pestis during infection and reveal the genetic patterns of the pathogen's survival strategy in different hosts.
Andrews
Gerard P
GP
Department of Veterinary Sciences, University of Wyoming, Laramie, Wyoming 82070, USA. gandrews@uwyo.edu
Vernati
Giulia
G
Ulrich
Ricky
R
Rocke
Tonie E
TE
Edwards
William H
WH
Adamovicz
Jeffrey J
JJ
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D017124:Conserved Sequence; D004269:DNA, Bacterial / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D010930:Plague / Q000382:microbiology*; D011817:Rabbits; D015010:Yersinia pestis / Q000235:genetics* / Q000302:isolation & purification*
D001426:Bacterial Proteins; D004269:DNA, Bacterial