Membrane glycoproteins of human polymorphonuclear leukocytes that act as receptors for mannose-specific Escherichia coli

Abstract

Type 1 fimbriated (mannose-specific) Escherichia coli cells bind to mannose residues on human polymorphonuclear leukocytes (PMN); this leads to phagocytosis of the bacteria. To identify the mannose-containing receptors on the PMN, the cells were surface labeled with 1251 and lysed in 0.5% Nonidet P-40, and the lysate was fractionated by affinity chromatography on a column of Sepharose-bound fimbriae. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography of the material eluted from the column with 500 mM methyl-a-mannoside revealed two radioactive bands of Mr 70,000 to 80,000 (gp7O-80) and 100,000 (gplOO). Another weak band of Mr 150,000 (gpl5O) was observed after prolonged exposure of the gel.Upon blotting of the glycoproteins separated by polyacrylamide gel electrophoresis and overlaying of the blots with concanavalin A, gpl50 appeared as the major band. Membrane preparations of the PMN were enriched in gp7O-80, gplOO, and gpl50, in comparison with the cell homogenates, further suggesting that these glycoproteins are surface components. Fractionation of the membrane preparations on the immobilized fimbriae followed by concanavalin A overlay of blots of the methyl-a-mannoside-eluted material revealed that gplSO was the major component in this fraction. The eluted fraction, obtained from a cell lysate (4.4 ,ug/ml), inhibited by 70% the agglutination of yeasts by the intact bacteria. Our results suggest that the three surface glycoproteins isolated by us serve as receptors for mannose-specific E. coli on PMN and may be involved in the lectin-mediated phagocytosis of the bacteria.

Klebsiellaspp. as Nosocomial Pathogens: Epidemiology, Taxonomy, Typing Methods, and Pathogenicity Factors

“…Once in the host tissue, however, the type 1 pili are no longer of use to the bacteria, since they trigger an opsonin-independent leukocyte activity known as lectinophagocytosis (167). The repulsion forces separating bacterium and leukocyte are weakened by the hydrophilic character of these pili (169), thus enabling the adhesins to bind to specific mannose-containing receptors on the leukocyte surface (205). Adhesin-binding triggers stimulation of the leukocyte (137), which ultimately leads to phagocytosis and intracellular killing of the bacterium (131).…”

Binding of Escherichia coli S fimbriae to cultured human endothelial cells

“…The role of bacterial adhesion in septicemia and hematogeneous infections is less clear, but there is evidence suggesting that it may be involved in the pathogenesis of neonatal sepsis and meningitis caused by Escherichia coli 018:K1 (9). These strains typically carry two types of adhesins, type 1 fimbriae that bind to oligomannoside chains (1,20) and S fimbriae that bind to sialylo2-3galactoside units of cell surface glycoproteins (17). 018:K1 E. coli expresses S fimbriae in blood during experimental septic infections in mice and infant rats, whereas type 1-fimbriated bacteria seem to be eliminated from circulation (11,21).…”

Identification of Polymorphonuclear Leukocyte and HL-60 Cell Receptors for Adhesins of Streptococcus gordonii and Actinomyces naeslundii

“…The 150-and 130-kDa glycoprotein bands on transfers of HL-60 or PMN cell extracts were not detected by ConA. Instead, ConA reacted with a distinct set of glycoproteins on both untreated and sialidase-treated blots, including one near 150 kDa that may serve as a major receptor for mannose-specific Escherichia coli (38). In control lanes, ConA reacted weakly with both fetuin and asialo-fetuin and strongly with thyroglobulin.…”