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Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis are Components of a Polymicrobial Intracellular Flora within Human Buccal Cells

¹ Department of Oral Science, School of Dentistry, and
² Biomedical Imaging and Processing Laboratory, Department of Neuroscience, University of Minnesota, 17-252 Moos Tower, 515 Delaware St. SE, Minneapolis, MN 55455, USA;

View larger version (91K): [in this window] [in a new window]   Figure 1. Panels A-C show images of z-section No. 39 from a stack of 74 0.2-µm z-sections (magnification 600X; the scale bar in Panel A also applies to Panels B and C). BEC in this field were double-labeled with the EUB338 universal probe (A) and the A. actinomycetemcomitans-specific probe (B). The cell in the center of Panel A contained a large mass of brightly fluorescent intracellular bacteria (red arrow). Other cells in the field contained smaller bacterial masses (not marked). Panel B shows that a portion of the large mass labeled with the universal probe also hybridized with the A. actinomycetemcomitans-specific probe (green arrow). The images from Panels A and B were superimposed (Panel C), confirming that bacteria labeled with both probes (yellow arrow) were adjacent to other bacteria labeled only with the universal probe (red arrow). Panel D presents a three-dimensional reconstruction of the same field. Bacteria recognized only by the universal probe are shown in solid red, while co-localization of the A. actinomycetemcomitans and universal probes is depicted by a green wireframe over a red interior. Reconstructed BEC surfaces are presented in blue. The red and green colors are muted when bacterial masses are intracellular, and brighter when bacteria appear to project out of the surface. The angle of view was rotated along the z-axis, and the image was zoomed. The large mass which appeared to have a lobular structure in z-section No. 39 was seen to be a cohesive unit containing A. actinomycetemcomitans in direct proximity to other species (red and green arrows).

View larger version (67K): [in this window] [in a new window]   Figure 2. In Panel A, the green wireframe denotes bacteria labeled by the P. gingivalis-specific probe. There were many bacterial masses within the cell shown, but only small portions of those masses appeared to contain P. gingivalis. In Panel B, the green wireframe indicates bacteria labeled by the T. forsythensis-specific probe. Note that the upper perimeter of that cell could not be properly rendered. The cell seemed to be almost completely filled with bacteria. Large masses of T. forsythensis were present, but species recognized only by the universal probe appeared to dominate the intracellular flora (the scale bar in Panel B applies to Panel A as well).

Journal of Dental Research, Vol. 84, No. 1, 59-63 (2005)
DOI: 10.1177/154405910508400110


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