Abstract
CD109 gene encodes a glycosylphosphatidylinositol-linked glycoprotein found in a subset of platelets and endothelial cell, and human platelet antigen (HPA) 15 is found on CD109. We evaluated the HPA genotype and/or the CD109 mRNA expression on two peripheral blood stem cells (PBSC), two peripheral bloods (PB), 12 granulocyte products, natural killer (NK)-92, B-lymphocyte (CO88BV59-1), K-562 leukemia cell line, human embryonic stem cell (hESC), and human fibroblasts (HF). HPA genotyping was performed by SNaPshot assay and CD109 mRNA expression was evaluated by real-time PCR with SYBR green and melting curve analysis. Genotype HPA-15a/-15a was found in PBSC#1 and two granulocyte products, and HPA-15a/-15b was found in PBSC#2, eight granulocyte products, NK-92, K-562, hESC, and HF, and HPA-15b/-15b was found in two granulocyte products. CD109 mRNA expression was highly increased in HF and increased in CD34+ and CD34− PBSCs and some granulocyte products, compared to the PB. However, the increase of expression level varied among the PBSC and granulocyte products. The CD109 mRNA expression of NK-92, K-562, hESC, and CO 88BV59-1 was not detected. HPA genotype was evaluated in various cells and the expression of CD109, which contains HPA 15, was different among cell lines and high in HF and PBSCs.
Highlights
The CD109 gene encodes a glycosylphosphatidylinositol linked glycoprotein, that is, a member of the alpha2macroglobulin/complement family of thioester-containing proteins [1]
Genotyping of human platelet antigen including HPA 15, which is located in CD109, was performed on different cells and cell lines
There have been no previous reports on the HPA genotype of cell lines, natural killer (NK)-92, CO 88BV59-1 Blymphocyte, K-562 leukemia, cell lines, and cells including human fibroblasts (HF) and human embryonic stem cell (hESC)
Summary
The CD109 gene encodes a glycosylphosphatidylinositol linked glycoprotein, that is, a member of the alpha2macroglobulin/complement family of thioester-containing proteins [1]. CD109 is expressed on platelets, activated Tcells, and endothelial cells [2] and has gained clinical attention due to the association of antibodies to CD109 with alloimmune thrombocytopenia and posttransfusion purpura [3]. Depending on a single nucleotide polymorphism at nucleotide of coding CD109 gene, human platelet antigen (HPA)-15a, and HPA-15b alleles are defined [4], and antiHPA-15 antibodies may develop and cause clinical consequences. CD109 mRNA transcript has been studied in various cancers and is found to be highly expressed in squamous cell carcinomas and melanomas [5–7]. The studies of the CD109 mRNA expression have been limited to few cell types we have evaluated mRNA expression in a wider variety of cell types and cell lineages including common cell lines. The cell types or cell lineages included in this study are peripheral blood stem cells (PBSC), granulocyte products, natural killer. The genotype of different HPAs including the HPA 15, which is included in the CD109, was characterized
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