Characterization of lipoteichoic acid binding to polymorphonuclear leukocytes of human blood

Abstract

Preparation of LTA. [3H]LTA (specific activity, 20,000 to 30,000 cpm/,ug) was obtained by growing streptococci in the presence of radiolabeled glycerol as previously described (19). Briefly, group A streptococci, strain 1RP41 (originally obtained from Rebecca Lancefield, The Rockefeller University, New York, N.Y.), were grown in 5-liter batches in van de Rijn-Kessler defined medium (26) containing 2.5 mCi of [2-3H]glycerol (specific activity, 200 mCi/mmol; New England Nuclear Corp., Boston, Mass.) per liter. After 16 h of growth at 370C, the bacteria were harvested

Stimulation of monokine production by lipoteichoic acids

“…LTAs have been reported to bind to, and in some cases influence the function of, several cell types, including epithelial cells (3,7,13), platelets (4), polymorphonuclear leucocytes (15), lymphocytes (5), and erythrocytes (6), and binding to granulocytes and monocytes apparently stimulates the respiratory burst (26,31). A requirement for fatty acids in mediating cell binding of LTA was previously shown (15,41). In the present study, a potential role for the negatively charged poly(glycerophosphate) chains was additionally detected by the use of polycations.…”

Effects of a streptococcal lipoteichoic acid on complement activation in vitro

“…Fragmentation of early components, e,g., C3, might deprive the phagocytes of opsonins, or saturate their C3 receptors. It is of interest that in vitro treatment of blood neutrophils with gingival fiuid conferred a binding defect on the cells' C3b receptors (Wilton et al 1977), Work by others showed that streptococcal LTA bound to receptor molecules on phagocytic cells (Levy et al 1990), and inhibited attachment of streptococci to neutrophils (Courtney et al 1981), Such LTAs might also biock, nonspecifically, Fc receptors on neutrophils (Ginsburg et al 1988). In any event, crevicular neutrophils from some forms of periodontitis seem to be associated with diminished phagocytic function, most likely induced by the environment of the lesion (Newman & Addison 1982).…”

Anti-lipoteichoic acid antibodies enhance release of cytokines by monocytes sensitized with lipoteichoic acid

“…In one study, it was found that LTA alone was not able to induce an oxidative burst in polymorphonuclear leukocytes, but upon treatment of the cells with anti-LTA antibodies, a brisk production of oxygen radicals was observed (8). Because LTA alone bound to polymorphonuclear leukocytes but was not capable of inducing patching and redistribution of LTA receptors unless anti-LTA was present (6), it was postulated that cross-linking of LTA receptors is required for production of oxygen radicals. Indeed, in human monocytes, the stimulation of the oxidative burst induced by LTA alone was associated with some degree of cross-linking of LTA receptors, as evidenced by patching and capping (12).…”

“…LTA from Streptococcus pyogenes M type 3 was extracted with hot phenol-water and purified by gel filtration chromatography as described previously (6,20). Deacylated LTA (d-LTA) was obtained by subjecting LTA to mild alkaline (0.2 M NaOH at 37°C) treatment for 60 min followed by neutralization with 0.1 M HCl (11).…”

“…LPS from Salmonella enteritidis, used as a positive control, was purchased from Difco (Diagnostic International Distribution, Milan, Italy). Anti-LTA serum was prepared by immunizing rabbits with LTA-methylated albumin complexes (6,22). Immunoglobulin G (IgG) was purified from anti-LTA serum by ion-exchange chromatography, using a quaternary aminoethyl Sephadex A-50 column equilibrated with 0.1 M Tris buffer, pH 6.5, as described previously (3).…”