Adherence of Streptococcus agalactiae to synchronously growing human cell monolayers without lipoteichoic acid involvement

Abstract

Freshly isolated virulent and nonvirulent strains of Streptococcus agalactiae type III were used to study differences in coccal adherence to synchronously dividing, subconfluent human embryonic amnion and fetal lung monolayers in vitro. The adherence frequency by virulent isolates of mid-logarithmically growing cocci to amnion cells varied markedly with host cell age, being highest shortly after eucaryotic cell division. This variation was not observed with lung cell monolayers, suggesting that cyclic production or exposure of coccal receptor sites on the eucaryotic cell surface with age is not a common property of all primary human cells in vitro. However, and regardless of age, not all cells within these synchronously dividing populations bound virulent cocci, indicating that a very small segment of a population may always be unresponsive to host cell interactions with a coccal pathogen. By comparison, adherence of nonvirulent coccal isolates to amnion and lung cells remained constant and of a very low order, regardless of host cell age. Maximal adherence of virulent S. agalactiae to young host cells occurred at early and mid-logarithmic phases of growth. However, at the late stationary growth phase, adherence was reduced to almost that of nonvirulent isolates. Pretreatment of virulent S. agalactiae with anti-lipoteichoic acid (LTA) serum failed to inhibit coccal adherence to these different host cells. Heat negated adherence. Group B coccal LTA was cytotoxic for these host cells. However, pretreatment of amnion and lung cells with nontoxic levels of this amphiphile did not prevent attachment of virulent cocci. Finally, coccal pretreatment with pronase abrogated adherence to either host cell even though surface-exposed LTA was uneffected, as observed by the indirect fluorescent-antibody procedure. Likewise, no observable difference in surface LTA was detected when fresh isolates of virulent and nonvirulent coccal strains were compared by this procedure. These studies suggest that protein involvement, rather than LTA, is primarily responsible for mediating virulent S. agalactiae type III attachment to these synchronously growing, subconfluent eucaryotic monolayers in vitro.