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Infection and Immunity, April 2003, p. 2199-2207, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.2199-2207.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Group A Streptococcus Gene Expression in Humans and Cynomolgus Macaques with Acute Pharyngitis

Kimmo Virtaneva,1 Morag R. Graham,1 Stephen F. Porcella,1 Nancy P. Hoe,1 Hua Su,1 Edward A. Graviss,2 Tracie J. Gardner,2 James E. Allison,3 William J. Lemon,4 John R. Bailey,5 Michael J. Parnell,5 and James M. Musser1*

Laboratory of Human Bacterial Pathogenesis,1 Veterinary Branch, Rocky Mountain Laboratories, National Institute of AllergyInfectious Diseases, National Institutes of Health, Hamilton, Montana 59840,5 Department of Pathology, Baylor College of Medicine, Houston, Texas 77030,2 Pediatric Medical Group, Houston, Texas 77098,3 Division of Human Cancer Genetics, Ohio State University Comprehensive Cancer Center, Columbus, Ohio 432104

Received 14 October 2002/ Returned for modification 10 December 2002/ Accepted 9 January 2003

The molecular mechanisms used by group A Streptococcus (GAS) to survive on the host mucosal surface and cause acute pharyngitis are poorly understood. To provide new information about GAS host-pathogen interactions, we used real-time reverse transcription-PCR (RT-PCR) to analyze transcripts of 17 GAS genes in throat swab specimens taken from 18 pediatric patients with pharyngitis. The expression of known and putative virulence genes and regulatory genes (including genes in seven two-component regulatory systems) was studied. Several known and previously uncharacterized GAS virulence gene regulators were highly expressed compared to the constitutively expressed control gene proS. To examine in vivo gene transcription in a controlled setting, three cynomolgus macaques were infected with strain MGAS5005, an organism that is genetically representative of most serotype M1 strains recovered from pharyngitis and invasive disease episodes in North America and Western Europe. These three animals developed clinical signs and symptoms of GAS pharyngitis and seroconverted to several GAS extracellular proteins. Real-time RT-PCR analysis of throat swab material collected at intervals throughout a 12-day infection protocol indicated that expression profiles of a subset of GAS genes accurately reflected the profiles observed in the human pediatric patients. The results of our study demonstrate that analysis of in vivo GAS gene expression is feasible in throat swab specimens obtained from infected human and nonhuman primates. In addition, we conclude that the cynomolgus macaque is a useful nonhuman primate model for the study of molecular events contributing to acute pharyngitis caused by GAS.

* Corresponding author. Mailing address: Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 903 South 4th St., Hamilton, MT 59840. Phone: (406) 363-9315. Fax: (406) 363-9427. E-mail: jmusser{at}niaid.nih.gov.

Editor: D. L. Burns

Infection and Immunity, April 2003, p. 2199-2207, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.2199-2207.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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