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Infection and Immunity, January 1999, p. 395-402, Vol. 67, No. 1
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

Annette Gasser, and Johannes Möst*

Institute of Hygiene, University of Innsbruck, Innsbruck, Austria

Received 24 June 1998/Returned for modification 21 July 1998/Accepted 15 October 1998

Multinucleated giant cells (MGC), a characteristic feature of tuberculous granulomas, form by fusion of monocytes or macrophages, but little is known about the mechanism of the fusion process itself. Several studies report an indirect effect of mycobacteria, i.e., induction of a soluble lymphocyte-derived fusion factor following stimulation by mycobacteria or mycobacterial products. The aim of our study was to determine whether contact with mycobacteria can induce MGC formation from human monocytes in vitro. Stimulation of monocytes with Mycobacterium bovis bacillus Calmette-Guérin (BCG) in combination with cytokine-containing supernatants of herpesvirus saimiri-transformed human T-cell clones (T-SN) led to MGC formation with fusion rates of about 27%. In contrast, stimulation with one component alone induced only low fusion rates of up to 10%. Heat-killed BCG in combination with T-SN induced monocyte fusion to the same extent as live mycobacteria. BCG culture supernatant, BCG lysate, or inert particles in combination with T-SN did not induce MGC formation. Experiments using transwell plates containing a semipermeable membrane revealed that induction of the fusion process is dependent on direct contact of monocytes and mycobacteria. MGC formation induced by BCG plus T-SN could be inhibited by addition of monoclonal antibodies to gamma interferon (but not tumor necrosis factor alpha) as well as to the beta  chain (CD18) of beta 2-integrins. These results demonstrate that contact with mycobacteria in combination with cytokine-containing supernatants is able to induce human monocytes to form MGC and that membrane-bound molecules of mycobacteria and monocytes are involved in the fusion process.

* Corresponding author. Mailing address: Institut für Hygiene, Fritz-Pregl-Str. 3, A-6020 Innsbruck, Austria. Phone: 43-512-507-3405. Fax: 43-512-507-2870. E-mail: johannes.moest{at}uibk.ac.at.

Infection and Immunity, January 1999, p. 395-402, Vol. 67, No. 1
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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