Cell-free released components of Streptococcus sanguis inhibit human platelet aggregation.
To study the role of surface components in the selective binding and aggregation of platelet-rich plasma (PRP) by strains of viridans streptococci, we treated the binding, aggregation strain Streptococcus sanguis I 2017-78 by sonication or trypsinization. Morphologically identifiable electron-dense fibrils were released from the cell wall, apparently from an inner electron-dense layer, under conditions that left cells intact. These controlled conditions were determined to cause submaximal loss in adhesion to platelet ghosts and PRP aggregation by treated, washed S. sanguis. Soluble components were recovered from the controlled sonic or L-(tosylamido 2-phenyl)ethyl chloromethyl ketone-trypsin treatments. Each showed dose-response inhibition of aggregation when preincubated with PRP before challenge with fresh, untreated S. sanguis. The time to onset of PRP aggregation was inhibited by 50% with 0.2 mg of TPCK-trypsin peptides or 1.0 mg of the sonicate per ml per 2 X 10(8) platelets. Components of both preparations were immunologically cross-reactive, but lipoteichoic acid was not a major antigen of either. By weight, the TPCK-trypsin peptides were virtually all protein; the sonicate residues identified were about 50% protein and 7% hexose. Each was a complex mixture of components as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. More than 8 TPCK-trypsin peptides and 16 sonicate components were so identified. In contrast, at least four or five components from either preparation were recognized as surface determinants by a rabbit antiserum to whole homologous microbes. Platelet-binding ligands of S. sanguis could be among these determinants.
D000268:Adhesiveness; D001792:Blood Platelets / Q000382:microbiology; D005690:Galactose / Q000032:analysis; D005944:Glucosamine / Q000032:analysis; D005947:Glucose / Q000032:analysis; D006801:Humans; D010455:Peptides / Q000032:analysis / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects*; D013010:Sonication; D013298:Streptococcus sanguis / Q000032:analysis* / Q000502:physiology; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
D010455:Peptides; D014108:Tosylphenylalanyl Chloromethyl Ketone; D014357:Trypsin; D005947:Glucose; D005944:Glucosamine; D005690:Galactose