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Infection and Immunity, December 2008, p. 5686-5693, Vol. 76, No. 12
0019-9567/08/$08.00+0     doi:10.1128/IAI.00133-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Mechanisms of T-Lymphocyte Accumulation during Experimental Pleural Infection Induced by Mycobacterium bovis BCG{triangledown}

Mariana C. Souza, Carmen Penido, Maria F. S. Costa, and Maria Graças Henriques*

Laboratório de Farmacologia Aplicada, Departamento de Farmacologia Aplicada, Farmanguinhos, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil

Received 30 January 2008/ Returned for modification 14 March 2008/ Accepted 14 September 2008

Tuberculous pleurisy is a frequent extrapulmonary manifestation characterized by accumulation of fluid and inflammatory cells in the pleural space. Here, we investigated the mechanisms of T-lymphocyte accumulation in the pleural space by using a murine model of pleurisy induced by Mycobacterium bovis BCG. Intrathoracic (i.t.) injection of BCG (4.5 x 105 bacteria/cavity) induced accumulation of T lymphocytes in the pleural cavities of C57BL/6 mice. We observed the presence of CFU in pleural washes conducted 1, 2, 3, 7, and 15 days after pleurisy induction. Pretreatment with fucoidan inhibited T-lymphocyte accumulation at 1 day, but not at 15 days, after BCG-induced pleurisy. Accordingly, adoptive transfer of fluorescein isothiocyanate-labeled blood mononuclear cells to infected mice showed that T lymphocytes migrated into the pleural cavity 1 day (but not 15 days) after BCG injection. Cell-free pleural wash fluids recovered from mice 1 day after BCG i.t. stimulation (day 1 BCG-PW), but not day 7 or day 15 BCG-PW, induced in vitro T-cell transmigration, which was dependent on L-, P-, and E-selectins. In contrast, day 7 BCG-PW (but not day 1 BCG-PW) induced in vitro T-lymphocyte proliferation via interleukin-2 (IL-2) and gamma interferon (IFN-{gamma}). Accordingly, in vivo IL-2 or IFN-{gamma} neutralization abolished T-lymphocyte accumulation 7 days after pleurisy induction. Our results demonstrate that pleural infection induced by BCG leads to T-lymphocyte accumulation in two waves. The acute phase depends on selectin-mediated migration, while the second wave of T-lymphocyte accumulation seems to depend on a local proliferation induced by cytokines produced in situ.

* Corresponding author. Mailing address: Laboratório de Farmacologia Aplicada, Departamento de Farmacologia Aplicada, Farmanguinhos, Fundação Oswaldo Cruz, Rua Sizenando Nabuco 100, Manguinhos, Rio de Janeiro 21041-250, Brazil. Phone: 55 21 3977-2478. Fax: 55 21 2564-2559. E-mail: gracahenriques{at}fiocruz.br

{triangledown} Published ahead of print on 22 September 2008.

Editor: J. L. Flynn

Infection and Immunity, December 2008, p. 5686-5693, Vol. 76, No. 12
0019-9567/08/$08.00+0     doi:10.1128/IAI.00133-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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