ArtinM: Purification and Evaluation of Biological Activities

Methods Mol Biol. 2020:2132:349-358. doi: 10.1007/978-1-0716-0430-4_34.

Abstract

The immunomodulatory activity of plant lectins has been evaluated because of their high selectivity for glycans linked to receptors on innate and adaptative immune cells. ArtinM is a mannosyl-binding lectin, obtained from the seeds of Artocarpus heterophyllus, that induces the differentiation of CD4+ T cells and macrophages by interacting with CD3 and TLR2/CD14, respectively. This ArtinM property ultimately favors the combat of intracellular pathogens, opening new perspectives on the lectins application as immunomodulatory agents. The current section describes protocols for purification and evaluation of ArtinM biological activity. The purification is based on the ArtinM-D-mannose affinity. The effect of inducing IL-12 production by murine macrophages cell line is adopted to evaluate the ArtinM biological activity.

Keywords: ArtinM; Immunomodulation; Innate and adaptative immune cells; Plant lectin; Purification.

MeSH terms

  • Animals
  • Artocarpus / chemistry
  • Artocarpus / metabolism*
  • CD3 Complex / metabolism
  • CD4-Positive T-Lymphocytes / cytology*
  • CD4-Positive T-Lymphocytes / drug effects
  • CD4-Positive T-Lymphocytes / metabolism
  • Cell Differentiation / drug effects
  • Cell Line
  • Immunologic Factors / isolation & purification
  • Immunologic Factors / pharmacology*
  • Interleukin-12 / metabolism
  • Lipopolysaccharide Receptors / metabolism
  • Macrophages / cytology*
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Mannose / metabolism
  • Mice
  • Plant Lectins / isolation & purification
  • Plant Lectins / pharmacology*
  • RAW 264.7 Cells
  • Seeds / chemistry
  • Seeds / metabolism
  • Toll-Like Receptor 2 / metabolism

Substances

  • CD3 Complex
  • Cd14 protein, mouse
  • Immunologic Factors
  • Lipopolysaccharide Receptors
  • Plant Lectins
  • Tlr2 protein, mouse
  • Toll-Like Receptor 2
  • Interleukin-12
  • Mannose