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The toxicity of lipoteichoic acid (LTA) from Streptococcus pyogenes type 12 was investigated by using mouse fibroblasts in culture in the absence of serum. Morphologically, while low concentrations of LTA elicited a subtle effect characterized by progressive cellular degeneration with practically no release of protein, larger concentrations (greater than 50 micrograms/ml) of this amphiphile resulted in rapid death of cell monolayers. Metabolic studies utilized a concentration of LTA (17.5 micrograms/ml) which caused the smallest change in cell morphology in the least number of mouse fibroblast cells per monolayer. Under these conditions, cell monolayers showed an increase of 450% in their content of collagenous protein after exposure to LTA. However, the amount of such material secreted remained unchanged. Also, changes in the type of collagenous protein formed were observed after exposure to LTA. Collagenous protein accumulating intracellularly was found to be practically hydroxyproline-free. However, collagenous protein secreted by this cell line showed a significantly reduced content of hydroxyproline as compared with control cells unexposed to this coccal membrane component. Column chromatographic studies confirmed that the collagenous protein secreted by monolayers exposed to LTA was defective (under hydroxylated). It was concluded that LTA does not affect the amount of collagenous protein secreted. However, it does increase the amount of this protein formed and retained by this cell line as well as causing a reduction in the hydroxylation of proline in both intracellular and secreted collagenous material. A possible relationship between abnormal basement membrane morphology and disturbed collagen synthesis in post-streptococcal glomerulonephritis as related to LTA is discussed.

Cytotoxicity and inhibition of normal collagen synthesis in mouse fibroblasts by lipoteichoic acid from Streptococcus pyogenes type 12