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Infection and Immunity, September 2005, p. 6026-6038, Vol. 73, No. 9
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.9.6026-6038.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification of Group A Streptococcus Antigenic Determinants Upregulated In Vivo

Kowthar Y. Salim,1 Dennis G. Cvitkovitch,1 Peter Chang,2 Darrin J. Bast,2 Martin Handfield,3 Jeffrey D. Hillman,3,4 and Joyce C. S. de Azavedo2*

Department of Microbiology, Faculty of Dentistry,1 Faculty of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada,2 Center for Molecular Microbiology and Department of Oral Biology, University of Florida College of Dentistry, Gainesville, Florida,3 iviGene Corp., Gainesville, Florida4

Received 30 November 2004/ Returned for modification 8 February 2005/ Accepted 19 May 2005

Group A Streptococcus (GAS) causes a range of diseases in humans, from mild noninvasive infections to severe invasive infections. The molecular basis for the varying severity of disease remains unclear. We identified genes expressed during invasive disease using in vivo-induced antigen technology (IVIAT), applied for the first time in a gram-positive organism. Convalescent-phase sera from patients with invasive disease were pooled, adsorbed against antigens derived from in vitro-grown GAS, and used to screen a GAS genomic expression library. A murine model of invasive GAS disease was included as an additional source of sera for screening. Sequencing DNA inserts from clones reactive with both human and mouse sera indicated 16 open reading frames with homology to genes involved in metabolic activity to genes of unknown function. Of these, seven genes were assessed for their differential expression by quantitative real-time PCR both in vivo, utilizing a murine model of invasive GAS disease, and in vitro at different time points of growth. Three gene products—a putative penicillin-binding protein 1A, a putative lipoprotein, and a conserved hypothetical protein homologous to a putative translation initiation inhibitor in Vibrio vulnificus—were upregulated in vivo, suggesting that these genes play a role during invasive disease.

* Corresponding author. Mailing address: Department of Microbiology, Mount Sinai Hospital, 600 University Ave., Rm. 1483, Toronto, Ontario M5G 1X5, Canada. Phone: (416) 586-8459. Fax: (416) 586-8746. E-mail: jdeazavedo{at}mtsinai.on.ca.

Editor: V. J. DiRita

Infection and Immunity, September 2005, p. 6026-6038, Vol. 73, No. 9
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.9.6026-6038.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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