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Infection and Immunity, October 2002, p. 5628-5634, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5628-5634.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Virulent Mycobacterium fortuitum Restricts NO Production by a Gamma Interferon-Activated J774 Cell Line and Phagosome-Lysosome Fusion

Tânia Regina Marques da Silva,1 Juliana Ribeiro de Freitas,1,2 Queilan Chagas Silva,1,2 Cláudio Pereira Figueira,1 Eliana Roxo,3 Sylvia Cardoso Leão,4 Luiz Antônio Rodrigues de Freitas,1,2 and Patrícia Sampaio Tavares Veras1,5*

Laboratório de Patologia e Biologia Celular, CPqGM, FIOCRUZ/BA,1 Faculdade de Medicina, Universidade Federal da Bahia,2 and Escola Bahiana de Medicina e Saúde Pública-SSA-BA, Bahia,5 Instituto Biológico, SP-SP, and,3 Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, UNIFESP/EPM, São Paulo, Brazil4

Received 1 March 2002/ Returned for modification 7 May 2002/ Accepted 9 July 2002

The virulence of different isolates of Mycobacterium has been associated with two morphologically distinguishable colonial variants: opaque (SmOp) and transparent (SmTr). In this report we used an in vitro assay to compare macrophage (M{phi}) responses to SmOp and SmTr Mycobacterium fortuitum variants, taking advantage of the fact that these variants were derived from the same isolate. Cells preactivated or not with gamma interferon (IFN-{gamma}) were infected with SmOp or SmTr M. fortuitum. We showed that SmOp and SmTr induced different levels of nitric oxide (NO) production by IFN-{gamma}-stimulated M{phi}. Indeed, the amount of IFN-{gamma}-induced NO production by J774 cells was 4.8 to 9.0 times higher by SmOp (23.1 to 37.7 µM) compared to SmTr infection (3.9 to 4.8 µM) (P = 0.0332), indicating that virulent SmTr bacilli restricted NO production. In addition, IFN-{gamma}-induced NO production by M{phi} was higher when correlated with reduction of only avirulent SmOp bacillus viability. SNAP (S-nitroso-N-acetyl-DL-penicillamine)-induced NO production did not modify SmTr viability, indicating its resistance to nitrogen radicals. Electron microscopy studies were performed to evaluate the capacity of phagosomes to fuse with lysosomes labeled with bovine serum albumin-colloidal gold particles. By 24 h postinfection, 69% more phagosome-containing SmOp variant had fused with lysosomes compared to the SmTr-induced phagosomes. In conclusion, these data indicate that virulent SmTr bacilli may escape host defense by restricting IFN-{gamma}-induced NO production, resisting nitrogen toxic radicals, and limiting phagosome fusion with lysosomes.

* Corresponding author. Mailing address: Laboratory of Pathology and Cellular Biology/FIOCRUZ BA R Valdemar Falcão, 121 Brotas Salvador, Bahia, Brazil 40295-001. Phone: 55-71-356-87-84. Fax: 55-71-356-42-92. E-mail: pstveras{at}hotmail.com.

Editor: J. D. Clements

Infection and Immunity, October 2002, p. 5628-5634, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5628-5634.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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