Histochemical Demonstration of Changes in Enzymatic Activity of KB Cells Produced by Staphylococcal Alpha and Beta Hemolysins

Abstract

Standarized suspensions of KB cells were incubated for 6 hr at 37[degree]C with various doses of purified alpha and beta hemolysins. Cells were observed for hematoxylin and Sudan B staining. Histochemical reactions for the following enzymes were performed thiamine pyrophosphatase/TPP-ase/, [alpha]-glutamyl-transpeptidase /GGTP-ase/, acid and alkaline phosphatases, adenosine triphosphatase /ATP-ase/, 5''-nucleotidase and nonspecific esterases /utilising as substrates beta-naphtyl-acetate and 5-bromo-indoxyl-acetate/. Enzyme activity was expressed in percentages of cells exhibiting presence of typical granulations. Application of various doses of both hemolysins, results in cytotoxic effect in higher and several alterations in cell enzymatic activities in lower concentrations. Difference in alterations caused by alpha and beta hemolysin, were noted. Alpha-hemolysin does not cause specific changes; only 5''-nucleotidase activity was affected. It is possible that this toxin acts on energetic systems of cells. Beta-hemolysin may disturb the lipid metabolism; this is supported partly by observation of increase in Sudan B staining lipids and decrease of activity of esterases. Both toxins do not exhibit activity of cholinesterase. Some implications of these results are discussed.