Specific binding of Haemophilus influenzae to minor gangliosides of human respiratory epithelial cells

Gangliosides are sialylated glycosphingolipids that serve as receptors for various bacteria, To investigate endogenous gangliosides of human respiratory epithelial cells as potential receptors for Haemophilus influenzae, three strains, including nontypeable H. influenzae (NTHI) 1479, and isogenic fimbriated (f(+)) and nonfimbriated (f(0)) H. influenzae type b 770235, were H-3 labeled and overlaid on two-dimensional thin-layer chromatography (TLC) plates containing either purified HEp-2 gangliosides or murine brain gangliosides: NTHI 1479 bound exclusively to two distinct minor ganglioside doublets, with mobilities near that of G(M1). These minor gangliosides comprised only 14.2 and 9.4% of the total, respectively, NTHI 1479 also bound to a distinct ganglioside of human macrophages whose chromatographic mobilities closely resemble those of one of the NTHI-binding gangliosides of HEp-2 cells, H. influenzae type b 770235 f(+) and f(0) each bound to a different minor HEp-2 ganglioside doublet, with proportionately weaker affinity for a major ganglioside doublet, Remarkably, none of the three strains bound to any murine brain gangliosides. Moreover, when 80 to 90% of sialic acid residues were enzymatically removed from HEp-2 gangliosides, NTHI 1479 binding was proportionately impaired, compared with untreated controls. Our findings support a role for specific gangliosides of specific cells as receptors for H. influenzae strains, Our findings further demonstrate that individual minor gangliosides possess unique biological properties.