The immunomodulatory β-galactoside-specific lectin from mistletoe: Partial sequence analysis, cell and tissue binding, and impact on intracellular biosignalling of monocytic leukemia cells

H. J. Gabius, H. Walzel, S. S. Joshi, J. Kruip, S. Kojima, V. Gerke, H. Kratzin, S. Gabius

Research output: Contribution to journalArticle

78 Scopus citations

Abstract

Nanogram quantities of the β-galactoside-specific lectin from mistletoe (ML-I) that is composed of two different types of subunits exhibit immunomodulatory potency and enhance cytokine secretion in vitro and in vivo. Partial sequence analysis of the carbohydrate-binding B chain revealed a ragged N-terminus and overall homologies to the B subunit of Ricin D and Ricin E. Two evolutionarily neutral substitutions were apparent in the otherwise identical N-terminal sequences of the two toxic chains within the lectin preparation. On the basis of the influence of chemical modification by group specific reagents on ligand binding, the lectin was biotinylated with biotinyl-N-hydroxysuccinimide ester to allow monitoring of cell binding. Monocytic leukemia cells (THP-1) specifically hound the lectin with positive cooperativity at low lectin concentrations. Radiolabelled lectin could be found in several organs and in an experimental solid tumor in biodistribution in mice. Its presence in a notable amount in spleens is especially noteworthy with respect to the already reported immunomodulation. To determine intracellular responses that precede the lectin-dependent augmentation of cytokine secretion, phosphorylation of proteins and phospholipids as well as Ca2+-mobilization were assessed in THP-1 cells. Quantitative increases of [32P]-phosphate incorporation were determined for a 28 kDa protein and for phosphatidylinositol-4,5-biphosphate. Similarly, the fluorescence activity of the intracellular Ca2+-indicator fluo-3 is elevated by approximately 25% after lectin treatment. Apparently, cell binding of the lectin is followed by modulation of biosignalling processes.

Original languageEnglish (US)
Pages (from-to)669-675
Number of pages7
JournalAnticancer Research
Volume12
Issue number3
StatePublished - 1992

Keywords

  • Biotinylation
  • Cytokine
  • Immunomodulation
  • Lectin
  • Protein phosphorylation

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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