Protective C allele of the single-nucleotide polymorphism rs1335532 is associated with strong binding of Ascl2 transcription factor and elevated CD58 expression in B-cells

Abstract

CD58 is expressed on the surface of antigen-presenting cells, including B-cells, and provides co-stimulation to regulatory T-cells (Treg) through CD2 receptor binding. Tregs appear to be essential suppressors of tissue-specific autoimmune responses. Thereby, CD58 plays protective role in multiple sclerosis (MS) and CD58 was identified among several loci associated with MS susceptibility. Minor (C) variant of the single-nucleotide polymorphism (SNP) rs1335532 is associated with lower MS risk according to genome-wide association studies (GWAS) and its presence correlates with higher CD58 mRNA levels in MS patients. We found that genomic region containing rs1335532 has enhancer properties and can significantly boost the CD58 promoter activity in lymphoblast cells. Using bioinformatics and pull-down assay we found that the protective (C) rs1335532 allele created functional binding site for ASCL2 transcription factor, a target of the Wnt signaling pathway. Both in B-lymphoblastoid cell lines and in primary B-cells, as well as in a monocytic cell line, activation of Wnt signaling resulted in an increased CD58 promoter activity in the presence of the protective but not the risk allele of rs1335532, whereas ASCL2 knockdown abrogated this effect. In summary, our results suggest that ASCL2 mediates the protective function of rs1335532 minor (C) allele in MS.

The Risk G Allele of the Single-Nucleotide Polymorphism rs928413 Creates a CREB1-Binding Site That Activates IL33 Promoter in Lung Epithelial Cells

“…NCIH-196 cells were transfected with siRNA duplexes (500 pmol of per 5 million cells) 48 h before transfection with the luciferase constructs and a further 200 pmol to extend the silencing effect. The measurement of CREB1 mRNA level by RT-PCR was performed as described [ 74 ]. Of three published pairs of siRNAs against CREB1 [ 75 ], one mediated a significant decrease in CREB1 mRNA level.…”

Multiomics dissection of molecular regulatory mechanisms underlying autoimmune-associated noncoding SNPs

“…These exclusive distal genes included many known immunologic genes, such as CD37 ( 45 ), CD28 ( 46 ), IL7 ( 47 ), IL12RB1 ( 48 ), or IL2RA ( 49 ). Previous functional assays have demonstrated that some distal immune-relevant genes could be regulated by distal enhancers in immune cell types, such as IL2RA ( 50 ), CD58 ( 51 ), IRF1 ( 52 ), or IRF5 ( 20 ), indicating the important roles of long-range regulation on autoimmune diseases. We further analyzed all 4,778 SNP-gene regulatory pairs and detected predominantly distal pairs (87.48%) compared with local genes ( Figure 5B and Supplemental Table 7 ).…”

Two genetic variants explain the association of European ancestry with multiple sclerosis risk in African-Americans

“…However, previously published experimental work has already begun to provide insight into the mechanism by which the CD58 gene variant contributes risk for MS. CD58, the protein coded for by the CD58 gene, is a cell adhesion molecule present on antigen presenting cells (APCs) that binds to CD2 on T cells to both strengthen the adhesion between T cells and APCs and to enhance T cell activation 24 . CD58 is also expressed in B cells, an important cell type in MS pathology, with higher levels of expression linked both to enhanced migration to inflammatory sites (for anti-inflammatory activity) and to CD2 ligation 25 . A past study showed that the protective allele of the rs2300474 variant, in strong LD with the protective allele of the rs12025416 variant, increases CD58 levels 17 (in contrast, the MS susceptibility haplotype, which harbors the risk allele of this variant, decreases CD58 expression).…”