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Infection and Immunity, July 2006, p. 4172-4179, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00447-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Proinflammatory and Immunomodulatory Cytokine mRNA Time Course Profiles in Hamsters Infected with a Virulent Variant of Leptospira interrogans

Frédérique Vernel-Pauillac and Fabrice Merien*

Laboratoire de Recherche en Bactériologie, Institut Pasteur de Nouvelle-Calédonie, 9-11 Avenue Paul Doumer, BP 61, 98845 Nouméa cedex, Nouvelle-Calédonie, France

Received 19 March 2006/ Returned for modification 17 April 2006/ Accepted 21 April 2006

In order to quantify in vivo the mRNAs of cytokines which play important roles in leptospirosis, we have developed quantitative real-time PCR assays for interleukin-2 (IL-2), IL-4, IL-10, IL-12p40, tumor necrosis factor alpha (TNF-{alpha}), gamma interferon (IFN-{gamma}), transforming growth factor ß, and two housekeeping genes (encoding ß-actin and hypoxanthine phosphoribosyltransferase). We used a lethal hamster model reflecting severe leptospirosis in humans. The LightCycler system was used to quantify the gene expression levels with the SYBR green I detection format using external standard curves for each target. We compared the expression levels of cytokine mRNA in the peripheral blood mononuclear cells of both control (uninfected) hamsters and Leptospira interrogans-inoculated hamsters from 1 to 24 h and then 1 to 4 days postinfection. In this kinetic study, there was pronounced expression of Th1 cytokine mRNA (TNF-{alpha}, IFN-{gamma}, and IL-12), with transcripts being detected as early as 1 h postinfection. Expression of anti-inflammatory cytokines, such as IL-4 and IL-10, was prominent in delayed samples from 1 to 4 days postinfection in response to infection with Leptospira interrogans. Our data are the first to establish that pathogenic leptospires can stimulate in vivo the production of type 1 cytokines involved in cellular immunity by using this informative animal model. Measuring and assessing cytokine profiles may provide a useful method for accurate study of the mechanisms of anti-Leptospira immunity, indications of prognosis factors, and prospective evaluation of leptospirosis vaccine efficacy in humans.

* Corresponding author. Mailing address: Institut Pasteur of New Caledonia, BP 61 98845 Noumea cedex, New Caledonia, France. Phone: 00 (687) 27 75 31. Fax: 00 (687) 27 33 90. E-mail: fmerien{at}pasteur.nc.

Editor: W. A. Petri, Jr.

Infection and Immunity, July 2006, p. 4172-4179, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00447-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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