Induction of L-variants in human diploid cells infected by group A streptococci

Human diploid cells in culture, infected with a balanced amount of living group A streptococci, were able to survive the infection and could be divided and propagated normally thereafter. The streptococci were rapidly phagocytized by the tissue culture cells. At the beginning, they kept their typical appearance, as well as their ability to fix dyes and group-specific immunoglobulins. After 1 to 2 days, the number of detectable streptococci decreased and they underwent important morphological changes. After some subsequent divisions of the cell line, streptococci persisted in cells only as large, isolated, swollen cocci, and no longer grew on suitable media. After six to eight divisions, a noticeable percentage of the tissue culture cells were very similar in appearance to the same cell line experimentally infected with "stable" L-variants. Cultures on L-phase media of supernatant fraction and cells, made 24 to 48 hr after inoculation, showed typical L-colonies. These grew well on media without antibiotics, as well as on media containing penicillin or vancomycin. They could be propagated on media with penicillin for months and were able to revert to group A streptococci after several subcultures on antibiotic-free media. Controls of uninoculated tissue culture cells never showed the presence of any microorganism. Group A streptococci inoculated into Eagle's basal medium, which was used for the tissue cultures, did not grow and never gave rise to L-colonies, even though the medium contained penicillin. Previous data suggest a biochemical explanation for this conversion, which otherwise is an occasional phenomenon.