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Infection and Immunity, July 2000, p. 4238-4244, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

In Vitro Cell Invasion of Mycoplasma gallisepticum

Florian Winner, Renate Rosengarten, and Christine Citti*

Institute of Bacteriology, Mycology and Hygiene, University of Veterinary Medicine Vienna, A-1210 Vienna, Austria

Received 3 February 2000/Returned for modification 4 April 2000/Accepted 17 April 2000

The ability of the widespread avian pathogen Mycoplasma gallisepticum to invade cultured human epithelial cells (HeLa-229) and chicken embryo fibroblasts (CEF) was investigated by using the gentamicin invasion assay and a double immunofluorescence microscopic technique for accurate localization of cell-associated mycoplasmas. The presence of intracellular mycoplasmas in both cell lines was clearly demonstrated, with organisms entering the eukaryotic cells within 20 min. Internalized mycoplasmas have the ability to leave the cell, but also to survive within the intracellular space over a 48-h period. Frequencies of invasion were shown to differ between the two cell lines, but were also considerably dependent on the mycoplasma input population. Of the prototype strain R, a low-passage population in artificial medium, Rlow, was capable of active cell invasion, while a high-passage population, Rhigh, showed adherence to but nearly no uptake into HeLa-229 and CEF. By passaging Rlow and Rhigh multiple times through HeLa-229 cells, the invasion frequency was significantly increased. Taken together, these findings demonstrate that M. gallisepticum has the capability of entering nonphagocytic host cells that may provide this pathogen with the opportunity for resisting host defenses and selective antibiotic therapy, establishing chronic infections, and passing through the respiratory mucosal barrier to cause systemic infections.

* Corresponding author. Mailing address: Institute of Bacteriology, Mycology and Hygiene, University of Veterinary Medicine Vienna, Veterinärplatz 1, A-1210 Vienna, Austria. Phone: 43 1 25077 2101. Fax: 43 1 25077 2190. E-mail: Christine.Citti{at}vu-wien.ac.at.

Infection and Immunity, July 2000, p. 4238-4244, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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