ABSTRACT
BACKGROUND & OBJECTIVES: Group A Streptococcus, causative agent of several clinical manifestations codes for multiple protein invasins which help the bacterium to enter non-phagocytic cells. C5a peptidase (SCPA) is a surface protein conserved among different serotypes of M1 strain. The present study was taken up to study SCPA promoted fibronectin independent entry of GAS into epithelial cells. METHODS: An isogenic 90226 emm1deltaAB (M1(-)) mutant was constructed with thermosensitive pGhost vector. This isogenic M1(-) mutant expressed SCPA on the surface as determined by Western blotting and immunofluorescence. RESULTS: On preincubation with anti-SCPA serum, the isogenic M1(-) strain exhibited 54 per cent decreased invasion as compared to the bacteria incubated with control serum. Also, purified recombinant SCPA proteins blocked internalization of M1(-) streptococci into HEp-2 cells. The M1(-) strain invaded at the same efficiency in the presence or absence of fibronectin. INTERPRETATION & CONCLUSION: These results suggested that SCPA acted as a potential invasin of group A streptococcus and promoted invasion independent of fibronectin.
Subject(s)
Adhesins, Bacterial/metabolism , Blotting, Western , Cell Line , Endopeptidases/metabolism , Epithelial Cells/microbiology , Fibronectins/physiology , Humans , Streptococcus pyogenes/physiologyABSTRACT
Strains of 12 T types of group A streptococci preserved by sand desiccation and stored at 4 degrees to 10 degrees C were regularly sub-cultured to check their viability. For this, streptococci coated onto sand particles mixed with sterile sheep blood were inoculated into Todd-Hewitt broth with added blood and incubated for 24 to 48 h. Checking for viability every six months showed that group A streptococci could be preserved by this method for 21 yr. Our study shows that sand desiccation is a convenient and cheap method for the long-term preservation of streptococci in laboratories where other methods of preservation are not feasible on a regular basis.
Subject(s)
Bacteriological Techniques , Cell Survival , Cryopreservation , Desiccation , Preservation, Biological/methods , Silicon Dioxide , Streptococcus pyogenes/physiologyABSTRACT
Lisogenia em amostras de estreptococos do grupo A foi examinada. Verificou-se que entre as lisogênicas a resistência a tetraciclina era mais comum