Abstract
To identify immunoglobulin variable heavy chain (VH) gene usages in Korean ankylosing spondylitis (AS) patients, expression level of VH2 genes from peripheral blood mononuclear cells (PBMCs) of 8 AS patients and 9 healthy donors was analysed by quantitative real-time PCR (Q-PCR). Q-PCR results demonstrated VH2 genes were overexpressed in AS patients (Relative amount of mRNA of VH2 genes to a house-keeping gene, 7.13+/-7.77 vs, 0.68+/-0.55; P<0.0001). The sequence analysis revealed the majority of them contained CDC42 binding protein kinase Beta (CDC42 BPB) genes. The insertion of CDC42 BPB gene was confirmed by PCR with primers corresponding CDC42 BPB and CH genes. Our study revealed VH2 overexpression and unique rearrangement in Ig VH genes from peripheral blood of AS patients. This may imply aberrant immunoglobulin gene rearrangement in B cell occurs in Korean AS patients, which requires further investigation.
Highlights
Ankylosing spondylitis (AS), a prototype of spondyloarthritis (SpA), is a chronic inflammatory arthritis that mainly affects the sarcroiliac joints and the spine
To analyze VH2 gene usages in detail, quantitative real-time PCR (Q-PCR) was performed with RNA from peripheral blood mononuclear cells (PBMCs) in AS patients
After we checked out that primer sets used in previous studies (Voswinkel et al, 2001) can cover variable heavy chain (VH) germline sequence from Ig Blast databases, we realized that 8 sequences of IGHV2 subfamilies (IGHV2-5*01, 5*04, 5*05, 5*06, 5*07, 5*10, 70*09 and 70*12) and 10 sequences of IGHV4 subfamilies (IGHV4-34*01, 34*03, 34*04, 34*05, 34*06, 34*07, 34*08, 34*11, 34*12 and 59*10) were not covered with those primer sets (Cowell et al, 1999; Van Esch et al, 2003) (Table 2)
Summary
Ankylosing spondylitis (AS), a prototype of spondyloarthritis (SpA), is a chronic inflammatory arthritis that mainly affects the sarcroiliac joints and the spine. It is characterized by peripheral arthritis, enthesitis, and extraskeletal features, such as uveitis and inflammatory bowel disease. Populations of immune cells have been implicated in the pathogenesis of AS/spondyloarthritis. Recent data have demonstrated that Th17 cells, IL-17-producing effector T helper cells, contribute to AS/ spondyloarthritis inflammation (Jandus et al, 2008; Shen et al, 2009). Specific autoantibodies have not yet been identified in patients with AS/spondyloarthritis, and the role or relevance of B cells in the pathogenesis of the disease is unclear
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