Nontypeable Haemophilus influenzae-binding gangliosides of human respiratory (HEp-2) cells have a requisite lacto/neolacto core structure.

Academic Article

Abstract

  • Nontypeable Haemophilus influenzae (NTHI) are a major cause of human infections. We previously demonstrated high affinity and high specificity binding of NTHI to minor gangliosides of human respiratory (HEp-2) cells and macrophages, but not to brain gangliosides. We further identified the NTHI-binding ganglioside of human macrophages as alpha2,3-sialylosylparagloboside (IV3NeuAc-nLcOse4Cer, nLM1), which possesses a neolacto core structure that is absent in brain gangliosides. This supported a hypothesis that lacto/neolacto core carbohydrates are critical for NTHI-ganglioside binding. To investigate, we determined the core carbohydrate structure of NTHI-binding gangliosides of HEp-2 cells, through multiple approaches, including specific enzymatic degradation, mass spectral analysis and gas-liquid chromatography. Our analyses denote the following critical structural attributes of NTHI-binding gangliosides: (1) a conserved lacto/neolacto core structure; (2) requisite sialylation, which may be either internal or external, with alpha2,3 (human macrophages) or alpha2,6 (HEp-2 cells) anomeric linkages; (3) internalized galactose residues. Mass spectral and gas chromatographic analyses confirm that NTHI-binding gangliosides of HEp-2 cells possess lacto/neolacto carbohydrate cores and identify the structure of the major peak as NeuAcalpha2-6Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glcbeta1-1Cer (alpha2,6-sialosylparagloboside, nLM1). Collectively, our studies denote NTHI-binding gangliosides as lacto/neolacto series structures.
  • Authors

  • Berenson, Charles S
  • Sayles, Kelly B
  • Huang, Jing
  • Reinhold, Vernon
  • Garlipp, Mary Alice
  • Yohe, Herbert C
  • Status

    Publication Date

  • August 1, 2005
  • Keywords

  • Bacterial Adhesion
  • Cell Line
  • Chromatography, Gas
  • Clostridium perfringens
  • Gangliosides
  • Haemophilus influenzae
  • Humans
  • Mass Spectrometry
  • Molecular Structure
  • Neuraminidase
  • Newcastle disease virus
  • Respiratory System
  • Structure-Activity Relationship
  • beta-Galactosidase
  • Digital Object Identifier (doi)

    Pubmed Id

  • 16051069
  • Start Page

  • 171
  • End Page

  • 182
  • Volume

  • 45
  • Issue

  • 2