Functional Evaluation of the Role of C-type Lectin Domain Family 16A at the Chromosome 16p13 Locus

Clin Exp Immunol. 2014 Mar;175(3):485-97. doi: 10.1111/cei.12240.

Abstract

The type 1 diabetes-associated 16p13 locus contains the CLEC16A gene. Its preferential immune cell expression suggests involvement in autoimmunity. Given its elevated expression in dendritic and B cells - known professional antigen-presenting cells (APCs) - we hypothesize that C-type lectin domain family 16 member A (CLEC16A) may be involved in T cell co-stimulation and consequent activation and proliferation. We also sought to identify CLEC16A's subcellular localization. The effect of the CLEC16A knock-down (KD) on B cell co-stimulation and activation of T cells was tested in human lymphoblastoid cell lines (LCLs) by co-culture with CD4(+) T cells. T cell activation and proliferation were determined by flow-cytometric analysis of CD69 and CD25 expression and carboxyfluorescein succinimidyl ester (CFSE) dilution, respectively. CLEC16A subcellular localization in K562 cells was examined by immunofluorescence. We show that the CLEC16A KD did not affect the tested indices of lymphoblastoid cell line (LCL) APC capacity. Additionally, the percentage of activated T cells following LCL co-culture was not affected significantly by the CLEC16A KD. T cells co-cultured with KD or control LCLs also exhibited similar cell division profiles. CLEC16A co-localized with an endoplasmic reticulum (ER) marker, suggesting that it may be an ER protein. In conclusion, CLEC16A may not be involved in T cell co-stimulation. Additional studies on CLEC16A, accounting for its ER localization, are needed to uncover its biological role.

Keywords: T cell activation and proliferation; co-culture assay; flow-cytometry; subcellular localization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigen-Presenting Cells / metabolism
  • B-Lymphocytes / metabolism
  • Chromosomes, Human, Pair 16*
  • Coculture Techniques
  • Endoplasmic Reticulum / metabolism
  • Gene Knockdown Techniques
  • Genetic Loci*
  • Humans
  • K562 Cells
  • Lectins, C-Type / genetics*
  • Lectins, C-Type / metabolism
  • Leukocytes, Mononuclear / metabolism
  • Lymphocyte Activation / immunology
  • Monosaccharide Transport Proteins / genetics*
  • Monosaccharide Transport Proteins / metabolism
  • Protein Transport
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism

Substances

  • CLEC16A protein, human
  • Lectins, C-Type
  • Monosaccharide Transport Proteins