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Infection and Immunity, April 2002, p. 1677-1683, Vol. 70, No. 4
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.4.1677-1683.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Peripheral Blood Mononuclear Cell Activation Induced by Leptospira interrogans Glycolipoprotein

Decio Diament,1,2 Milena Karina Colo Brunialti,1 Eliete Calo Romero,3 Esper Georges Kallas,1 and Reinaldo Salomao1*

Escola Paulista de Medicina, Federal University of São Paulo,1 Instituto de Infectologia Emilio Ribas,2 Instituto Adolpho Lutz, São Paulo, Brazil3

Received 18 May 2001/ Returned for modification 14 August 2001/ Accepted 3 January 2002

Leptospira interrogans glycolipoprotein (GLP) has been implicated in pathological and functional derangement seen in leptospirosis. The goal of this study was to evaluate GLP's ability to induce cellular activation, as assessed by cytokine production and expression of surface activation markers. GLP extracted from either pathogenic L. interrogans serovar Copenhageni or nonpathogenic Leptospira biflexa serovar Patoc (GLPp) was used to stimulate peripheral blood mononuclear cell cultures from healthy donors. Supernatant cytokine levels were measured by enzyme-linked immunosorbent assay. Expression of CD69 and HLA-DR on lymphocytes and monocytes, as well as lipopolysaccharide (LPS) binding, were measured by flow cytometry. At 6 h of incubation, GLP induced a significant rise in tumor necrosis factor alpha levels, which dropped progressively until 72 h of incubation. Interleukin-10 peak levels were obtained at between 24 and 48 h, with sustained levels until 72 h of incubation. The response magnitude was proportional to the GLP dose. CD69 expression on T lymphocytes and monocytes increased significantly, as did HLA-DR expression on monocytes. GLPp induced no CD69 or HLA-DR expression. GLP did not block biotinylated LPS binding to monocytes, suggesting that different pathways are used to induce cell activation. In conclusion, GLP induces cellular activation and may play a major role in the pathogenesis of leptospirosis.

* Corresponding author. Mailing address: R. Pedro de Toledo, 781, 15° andar 04039-032, Sao Paulo, SP, Brazil. Phone: 55-11-55798226. Fax: 55-11-50815394. E-mail: rsalomao-dipa{at}pesquisa.epm.br.

Editor: R. N. Moore

Infection and Immunity, April 2002, p. 1677-1683, Vol. 70, No. 4
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.4.1677-1683.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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