Summary
The host-parasite relationship of HeLa M cells artificially infected with a bovine species of Mycoplasma was studied by light microscopy, transmission electron microscopy and scanning electron microscopy. The use of morphometry to quantitate some of the findings was explored. The parasites were seen in locations extracellular to the cell surface. The detection of small numbers of organisms by light microscopy was well demonstrated by use of the fluorescent antibody technique. Scanning electron microscopy proved to be an excellent method for revealing the surface details of cell-parasite morphology. Ultra-thin sections showed that the parasites are aligned mostly parallel to the plasma membrane of the host cell but separated by a gap of 10 nm. Morphometry indicated an average of 69 organisms per cell surface occupying 1.7% of the surface area. An increase of 26% in diameter of the HeLa cells, possibly as a result of infection, was observed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ambrose, E.J.: Electrophoretic behavior of cells. Progr. Biophys. molec. Biol. 16, 243–265 (1966)
Barile, M.F., Malizia, W.F., Riggs, D.B.: Incidence and detection of pleuropneumoniae-like organisms in cell cultures by fluorescent antibody and cultural procedures. J. Bact. 84, 130–136 (1962)
Boatman, E.S.: Morphology and ultrastructure of mycoplasmas. Ann. N.Y. Acad. Sci. 225, 172–180 (1973)
Brown, S., Teplitz, M., Revel, J.P.: Interaction of mycoplasmas with cell cultures, as visualized by electron microscopy. Proc. nat. Acad. Sci. (Wash.) 71, 464–468 (1974)
Collier, A.M.: Pathogenesis of Mycoplasma pneumoniae infection as studied in the human foetal trachea in organ culture. In: Pathogenic mycoplasmas. New York: Associated Scientific Publishers 1972
Fogh, J., Fogh, H.: A method for direct demonstration of pleuro-pneumoniae-like organisms in cultured cells. Proc. Soc. exp. Biol. (N.Y.) 117, 899–901 (1964)
Fogh, J., Fogh, H.: Morphological and quantitative aspects of Mycoplasma — Human cell relationships. Proc. Soc. exp. Biol. (N.Y.) 125, 423–430 (1967)
Harnett, G.B., Phillips, P.A., Mackay-Scollay, E.M.: A simple method for detecting mycoplasma infection of cell cultures. J. clin. Path. 27, 70–73 (1974)
Hayflick, L.: Mycoplasmas as pathogens. In: Pathogenic mycoplasmas, pp. 17–37. New York: Associated Scientific Publishers 1972
Jones, T.C., Hirsch, J.G.: The interaction in vitro of Mycoplasma pulmonis with mouse peritoneal macrophages and L-cells. J. exp. Med. 133, 231–259 (1971)
Kenny, G.E.: Heat-lability and organic solvent-solubility of Mycoplasma antigens. Ann. N.Y. Acad. Sci. 143, 676–681 (1967)
Kenny, G.E.: Contamination of mammalian cells in culture with mycoplasmata. In: Contamination in tissue culture (J. Fogh, ed.), pp. 107–129. New York: Academic Press 1975
Kenny, G.E.: Serological heterogeneity of the Mycoplasmatales. J. infect. Dis. 127, S2-S5 (1973a)
Kenny, G.E.: Rapid detection of mycoplasmata and non-culturable agents in animal cell cultures by uracil incorporation. In: Microbiology (D. Schlessinger, ed.), pp. 32–36. Washington, D.C.: American Society for Microbiology 1975
Kenny, G.E.: The antigens of the Mycoplasmatales and Chlamydiae. In: The Antigens, Vol. III (M. Seala, ed.), pp. 449–478. New York: Academic Press 1975a
Lemcke, R.M.: Osmolar concentration and fixation of mycoplasmas. J. Bact. 110, 1154–1162 (1972)
Luft, J.H.: Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9, 409–416 (1961)
Manchee, R.J., Taylor-Robinson, D.: Studies on the nature of receptors involved in attachment of tissue culture cells to mycoplasmas. Brit. J. exp, Path. 50, 66–75 (1969)
McGarrity, G.J., Coriell, L.L.: Detection of anaerobic mycoplasmas in cell cultures. In vitro 9, 17–18 (1973)
Revel, J.P.: Scanning electron microscope of cell surface morphology and labeling in situ and in vitro. S.E.M. 1974. Proc. 7th Annual IIT Res. Inst. Chicago (Ill.), pp. 541–547 (1974)
Schneider, E.L., Stanbridge, E.J., Epstein, C.J.: Incorporation of 3H-uridine and 3H-uracil into R.N.A.: A simple technique for detection of mycoplasma contamination of cultured cells. Exp. Cell Res. 84, 311–318 (1974)
Sobeslovsky, O., Prescott, B., Chanock, R.M.: Adsorption of Mycoplasma pneumoniae to neuraminic acid receptors of various cells and possible role in virulence. J. Bact. 96, 695–705 (1968)
Wiebel, E., Bolender, R.: Stereological techniques for electron microscopic morphometry. In: Principles and techniques of electron microscopy, Vol. 3 (M.A. Hayat, ed.). New York: Van Nostrand Reinhold 1973
Author information
Authors and Affiliations
Additional information
The authors wish to thank Christiana Ulness and Andrea Erickson for expert technical assistance and Arnold Schmidt for the operation of the scanning electron microscope. This work was supported by grants from the U.S.P.H.S.: AI 09586, AI 10743, and AI 06720
Rights and permissions
About this article
Cite this article
Boatman, E., Cartwright, F. & Kenny, G. Morphology, morphometry and electron microscopy of HeLa cells infected with bovine Mycoplasma . Cell Tissue Res. 170, 1–16 (1976). https://doi.org/10.1007/BF00220107
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00220107