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Infection and Immunity, August 2005, p. 4505-4511, Vol. 73, No. 8
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.8.4505-4511.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Human ß-Defensin 2 Is Expressed and Associated with Mycobacterium tuberculosis during Infection of Human Alveolar Epithelial Cells

Bruno Rivas-Santiago,1,2 Stephan K. Schwander,3 Carmen Sarabia,2 Gill Diamond,4 Marcia E. Klein-Patel,4,5 Rogelio Hernandez-Pando,6 Jerrold J. Ellner,3 and Eduardo Sada2*

Department of Experimental Pathology, Centro de Investigación y Estudios Avanzados del IPN, Mexico City, Mexico,1 Department of Microbiology, Instituto Nacional de Enfermedades Respiratorias, Mexico City, Mexico,2 Department of Medicine and Center for Emerging Pathogens,3 Department of Oral Biology,4 GSBS, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103,5 Department of Pathology, Instituto Nacional de Ciencias Medicas SZ, Mexico City, Mexico6

Received 12 October 2004/ Returned for modification 5 December 2004/ Accepted 8 March 2005

To determine the role of human ß-defensin 2 (HBD-2) in human tuberculosis, we studied the in vitro induction of HBD-2 gene expression by Mycobacterium tuberculosis H37Rv infection in the human lung epithelial cell line A549, in alveolar macrophages (AM), and in blood monocytes (MN) by reverse transcription-PCR. We also studied the induction of HBD-2 gene expression by mannose lipoarabinomannan (manLAM) from M. tuberculosis. Intracellular production of HBD-2 peptide was detected by immunocytochemistry and electron microscopy. Our results demonstrated that there was induction of HBD-2 mRNA in A549 cells after infection with M. tuberculosis at various multiplicities of infection (MOI) and that there was stimulation with manLAM. AM expressed the HBD-2 gene only at a high MOI with M. tuberculosis. MN did not express HBD-2 at any of the experimental M. tuberculosis MOI. Immunostaining revealed the presence of intracellular HBD-2 peptide in A549 cells following infection with M. tuberculosis, and the staining was more intense in areas where there were M. tuberculosis clusters. By using electron microscopy we also demonstrated production of HBD-2 after M. tuberculosis infection and adherence of HBD-2 to the membranes of M. tuberculosis. Alveolar epithelial cells are among the first cells to encounter M. tuberculosis following aerogenic infection. As HBD-2 has been shown to control growth of M. tuberculosis and has chemotactic activity, our results suggest that HBD-2 induction by M. tuberculosis may have a role in the pathogenesis of human tuberculosis.

* Corresponding author. Mailing address: Department of Microbiology, Instituto Nacional de Enfermedades Respiratorias, Mexico City, Mexico. Phone: 525 556 666 172. Fax: 525 556 666 172. E-mail: eduardosasadiaz{at}yahoo.com.

Editor: J. L. Flynn

Infection and Immunity, August 2005, p. 4505-4511, Vol. 73, No. 8
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.8.4505-4511.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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