Immune labeling of certain strains of Actinomyces naeslundii and Actinomyces viscosus by fluorescence and electron microscopy

Abstract

A total of 12 well-characterized strains of Actinomyces viscosus and A. naeslundii grown on Trypticase soy agar plates supplemented with sheep erythrocytes were examined by light microscopy and transmission electron microscopy after treatment with appropriately labeled antisera to homologous and heterologous strains. Cells incubated with homologous rabbit antisera followed by fluorescein-isothiocyanate (FITC)-conjugated goat anti-rabbit immunoglobulin G (IgG) exhibited a completely smooth fluorescent outline in the case of A. naeslundii and and interrupted, irregular fluorescent outline in the case of human strains of A. viscosus. The different labeling patterns appeared to be related to the presence at the ultrastructural level of long, unevenly distributed strands of "fuzz" on the surface of human A. viscosus cells, whereas A. naeslundii cells had a narrower layer of fuzz, or more even thickness. The immunocoating reaction revealed homologous antibody binding to the irregular strands of fuzz on the surface of human A. viscosus cells, whereas homologous antisera to A. naeslundii coated A. naeslundii cells with a moderately electron-dense coating of antibody of even thickness. Human strains of A. viscosus incubated with heterologous antiserum to A. naeslundii followed by FITC-labeled goat anti-rabbit IgG exhibited a segmented fluorescent outline, which differed from that produced with homologous antisera. A. naeslundii incubated with heterologous rabbit antisera to human A. viscosus strains and FITC-labeled anti-rabbit IgG exhibited a completely smooth fluorescent outline similar to that produced with homologous antiserum. A. viscosus strains of hamster origin differed from A. viscosus strains of human origin by the absence of a surface fuzz and the comparatively smooth, even fluorescence produced by incubating these cells with homologous rabbit antiserum followed by FITC-labeled goat anti-rabbit IgG. Antiserum to a hamster strain did not cross-react with A. naeslundii or human strains of A. viscosus. Under the growth conditions of this experiment, ultrastructural features and labeling patterns with the indirect fluorescent technique may be useful in differentiating these serotypes from one another.