Interaction of Bartonella henselae with the Murine Macrophage Cell Line J774: Infection and Proinflammatory Response

doi:10.1128/IAI.69.10.5974-5980.2001

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Infection and Immunity, October 2001, p. 5974-5980, Vol. 69, No. 10
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.10.5974-5980.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Interaction of Bartonella henselae with the Murine Macrophage Cell Line J774: Infection and Proinflammatory Response

Tiziana Musso,¹^,^* Raffaele Badolato,² Daniela Ravarino,¹ Sarah Stornello,¹ Patrizia Panzanelli,³ Chiara Merlino,¹ Dianella Savoia,⁴ Rossana Cavallo,¹ Alessandro Negro Ponzi,¹ and Mario Zucca⁴

Department of Public Health and Microbiology,¹ Department of Anatomy, Pharmacology, and Forensic Medicine,³ and Department of Clinical and Biological Sciences,⁴ University of Turin, Turin, and Department of Pediatrics, University of Brescia, Brescia,² Italy

Received 12 March 2001/Returned for modification 4 May 2001/Accepted 10 July 2001

Bartonella henselae is the causative agent of cat scratch disease (CSD), a self-limiting condition characterized by a subacuteregional lymphadenopathy that may develop into disseminated bartonellosisin immunocompromised subjects. Mice experimentally infected withB. henselae display typical liver and spleen granulomas rich inT cells and macrophages. So far there are no data on the interactionbetween bartonellae and macrophages. In order to clarify thistopic, we investigated the interaction of B. henselae with J774,a mouse macrophage cell line. Analysis of bacterial uptake byfunctional assays and transmission electron microscopy indicatesthat bartonellae can enter and survive inside J774. Entry occurredwithin 30 min postinfection and reached a plateau at 160 min.Infection of J774 was followed by a dose-dependent release ofthe proinflammatory cytokines tumor necrosis factor alpha, interleukin1 $beta$ (IL-1 $beta$ ), and IL-6. Bartonellae persisted intracellularly withoutloss of viability for at least 8 h, and their number slightlydecreased 24 h postinfection. Gamma interferon (IFN- $gamma$ ) treatmentof J774 significantly decreased the number of recoverable bacteriaat 8 and 24 h. This enhancement of macrophage bactericidal activitywas associated with nitric oxide (NO) release and was preventedby the addition of the competitive inhibitor of NO synthesis N^G-monomethyl L-arginine. These findings suggest that IFN- $gamma$ -mediatedactivation of macrophages may be important for the clearing ofB. henselae infection and that anti-B. henselae microbicidal activityof IFN- $gamma$ -activated macrophages is mediated to a large extent byNOproduction.

^* Corresponding author. Mailing address: Istituto di Microbiologia, Via Santena 9, 10126 Torino, Italy. Phone: 39 011 670.6609.Fax: 39 011 663.6436. E-mail: tiziana.musso{at}unito.it.

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