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Recently, we reported that conjugal transfer of plasmid pERL1, determining i.a. erythromycin resistance (Emr), into group A streptococci could trigger the expression of anti-phagocytic activity, adhesiveness, opacity factor and capacity to bind immunoglobulin Fc-parts and beta 2-microglobulin. In the present study, ethidium bromide treatment of Emr transconjugants allowed the selection of "cured", erythromycin sensitive (Ems) mutants. This procedure did not affect the expression of the abovementioned characteristics. However, when plasmid pERL1 was again transferred to two such mutants, the "secondary", Emr transconjugants obtained showed lack of each of these properties. Our experiments thus demonstrated a "switch-on" as well as a "switch-off" effect, exerted by the same plasmid, pERL1, on some major pathogenic properties of group A streptococci.

Summary Submicroscopic aspects of the adherence of Group A streptococci to HEp-2 cells and the time sequence of their further interaction with these cells were studied. The M+ variant of streptococci, characterized by the presence of filamentous protrusions on the cell wall, displayed a high capacity for adherence, in contrast to the M- variant of the same strain, where adherence was low. The first stage of the interaction between M+variant of Group A streptococci and HEp-2 cells was adherence of the filamentous protrusions of the bacterial cell wall to host cell cytoplasmic membrane; this was followed by closer contact of the streptococcus cell wall with HEp-2 cell surface. Continuing incubation led to the development of invaginations in the cytoplasmic membranes of HEp-2 cells, into which streptococci gradually penetrated. Ingestion of streptococci into the forming pseudovacuoles of the host cell was accompanied by bacterial cell division, culminating in total disintegration of the host cell and release of the streptococci into the medium. At all stages of the interaction there was a pronounced tendency to form multiple contacts between the surface structures of the streptococcus cell and the membrane structures of the animal cell being attacked.

In vitro tests for adherence to human vaginal and pharyngeal epithelial cells were used to study the problem of tissue-specific tropism in group B streptococci (GBS). Twenty-two vaginal or pharyngeal clinical isolates of GBS (serotypes Ia, Ib, II, and III) were used. No significant differences in adherence to vaginal and pharyngeal epithelial cells were found between GBS from both sources: statistical analysis furnished no evidence for tissue-specific tropism. Serotype III vaginal GBS adhered better to vaginal and pharyngeal epithelial cells than did serotype III GBS strains isolated from the pharynx. However, pronounced differences in the level of adherence were found among strains of the same serotypes and from the same sources. Thus, the results obtained suggest that differences in adherence may rather be strain-dependent that type-dependent.

The adherence of group B streptococci (GBS) of serotypes Ia, II and III to human vaginal cells was studied in vitro. The adherence was not dependent on the viability of bacteria; killing of GBS by UV irradiation or glutaraldehyde treatment did not inhibit the adherence. Killing of GBS by heating to 56 degrees C for 1 h led to a pronounced decrease of adherence, demonstrating the thermosensitivity of the GBS structures involved. The protein nature of these structures was proved by a significant reduction of adherence after pretreatment of GBS with trypsin or pepsin. Pretreatment of GBS with sialidase had no influence on the adherence. Such a pretreatment of vaginal cells caused an increase of adherence showing that the receptors on epithelial cells may be partly masked by sialic acid.

Adherence of three strains of group A streptococci and their fosfomycin-resistant mutants to HEp-2 tissue culture cells was compared with some cell-surface characteristics, i.e. ultrastructure and hydrophobicity. Among Fosr mutants, both well-adhering and weakly adhering mutants were found. Clonal analysis of the mutants proved their greater stability in the adherence. Well-adhering parent strains of streptococci and Fosr mutants exhibited surface fibrillae in contrast to weakly adhering Fosr mutants which were devoid of fibrillae or contined fibrillae of lower electron density. Decrease of adherence of Fosr mutants of two strains was accompanied by a decrease of their hydrophobicity.