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Journal of Virology, September 2004, p. 9854-9861, Vol. 78, No. 18
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.18.9854-9861.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Coxsackievirus B4 Infection of Human Fetal Thymus Cells

Fabienne Brilot,1,{dagger} Vincent Geenen,2 Didier Hober,3* and Cheryl A. Stoddart1

Gladstone Institute of Virology and Immunology, University of California, San Francisco, California,1 Center of Immunology, Institute of Pathology, University of Liege, Liege-Sart-Tilman, Belgium,2 UPRES EA3610 Pathogenèse Virale du Diabète de Type 1, Faculté de Médecine et Service de Virologie, Université Lille 2, CHRU, Lille, France3

Received 26 February 2004/ Accepted 17 May 2004

The infection of human fetal thymus organ cultures (FTOC) with coxsackievirus B4 E2 (CVB4 E2) was investigated. Both positive- and negative-strand viral RNA were detected by real-time quantitative reverse transcription-PCR (RT-PCR) in CVB4 E2-infected FTOC, which supported high yields of virus production (~106 50% tissue culture infective doses/ml), and in flow-sorted thymocyte populations for 7 days after inoculation. Cortical CD4+ CD8+ thymocytes were found to be the principal targets of infection. Inoculation of human FTOC with CVB4 E2 led to a marked and progressive depletion of immature thymocytes (CD4+ CD8+ cells) with no enhancement of Annexin V-positive cells. CVB4 E2 replication caused significant major histocompatibility complex (MHC) class I upregulation on these cells. MHC class I upregulation was correlated with positive- and negative-strand RNA quantitative detection and the release of infectious particles. In addition, chloroquine treatment of FTOC and single-thymocyte suspensions suggested that MHC class I upregulation on thymocytes was the result of direct infection rather than caused by production of soluble factors such as alpha interferon. Thus, CVB4 E2 can infect human fetal thymocytes, which subsequently results in quantitative and qualitative abnormalities of these cells.

* Corresponding author. Mailing address: UPRES EA3610 Pathogenèse Virale du Diabète de Type 1, Université de Lille 2, Faculté de Médecine et Service de Virologie, BÂt. Paul Boulanger, CHRU, 59037 Lille, France. Phone: 33-3-20-44-66-88. Fax: 33-3-20-44-52-81. E-mail: dhober{at}chru-lille.fr.

{dagger} Present address: University of Liege, Center of Immunology, Institute of Pathology, Liege-Sart-Tilman, Belgium.

Journal of Virology, September 2004, p. 9854-9861, Vol. 78, No. 18
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.18.9854-9861.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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