title: HLA-DRB1 shared epitope genotyping using the revised classification and its association with circulating autoantibodies, acute phase reactants, cytokines and clinical indices of disease activity in a cohort of South African rheumatoid arthritis patients.
abstract: BACKGROUND
  The revised shared epitope (SE) concept in rheumatoid arthritis (RA) is based on the presence (S) or absence (X) of the SE RAA amino acid motif at positions 72 to 74 of the third hypervariable region of the various human leucocyte antigen (HLA)-DRB1 alleles. The purpose of this study was to investigate SE subtypes on the basis of the American College of Rheumatology 1987 revised criteria for the classification of RA in a cohort of South African RA patients (n = 143) and their association with clinical and circulating biomarkers of disease activity (autoantibodies, acute phase reactants and cytokines).
  
  
  METHODS
  Genomic DNA was analysed using high-resolution recombinant sequence-specific oligonucleotide PCR typing of the HLA-DRB1 allele. Subtypes of the SE were classified according to the amino acids at positions 72 to 74 for the RAA sequence, and further sub-divided according to the amino acids at positions 70 and 71, which either contribute to (S2, S3P), or negate (S1, S3D) RA susceptibility. Disease activity was assessed on the basis of (1) Disease Activity Score in 28 joints using C-reactive protein (CRP), (2) rheumatoid factor (RF), (3) CRP and (4) serum amyloid A by nephelometry, anticyclic citrullinated peptide antibodies (aCCP) by an immunofluorometric procedure, and cytokines by multiplex bead array technology.
  
  
  RESULTS
  Of the 143 RA patients, 81 (57%) were homozygous (SS) and 50 (35%) were heterozygous (SX) for the SE alleles with significant overexpression of S2 and S3P (respective odds ratios (ORs) 5.3 and 5.8; P < 0.0001), and 12 (8%) were classified as no SE allele (XX). Both the SS and SX groups showed a strong association with aCCP positivity (OR = 10.2 and P = 0.0010, OR = 9.2 and P = 0.0028, respectively) relative to the XX group. Clinical scores and concentrations of the other biomarkers of disease activity (RF, CRP and T helper cell type 1 (Th1), Th2, macrophage and fibroblast cytokines) were also generally higher in the SS group than in the SX and XX groups.
  
  
  CONCLUSIONS
  RA susceptibility alleles investigated according to revised criteria for the classification of RA were significantly increased in South African RA patients and strongly associated with aCCP in particular as well as with circulating cytokines and disease severity.
authors: Medical Research Council Unit for Inflammation and Immunity, Department of Immunology, Faculty of Health Sciences, University of Pretoria, and Tshwane Academic Division of the National Health Laboratory Service, Bophelo Road, Pretoria, 0001, South Africa. pieter.meyer@nhls.ac.za
authors: Pieter W A
authors: PW
authors: Meyer
authors: Bridget
authors: B
authors: Hodkinson
authors: Mahmood
authors: M
authors: Ally
authors: Eustasius
authors: E
authors: Musenge
authors: Ahmed A
authors: AA
authors: Wadee
authors: Heidi
authors: H
authors: Fickl
authors: Mohammed
authors: M
authors: Tikly
authors: Ronald
authors: R
authors: Anderson
mesh_terms: D000209:Acute-Phase Proteins / Q000378:metabolism*; D000328:Adult; D000368:Aged; D001172:Arthritis, Rheumatoid / Q000097:blood* / Q000453:epidemiology / Q000235:genetics*; D001323:Autoantibodies / Q000097:blood*; D015415:Biomarkers / Q000097:blood; D015331:Cohort Studies; D016207:Cytokines / Q000097:blood*; D004198:Disease Susceptibility / Q000097:blood; D018984:Epitopes, T-Lymphocyte / Q000235:genetics; D005260:Female; D059811:HLA-DRB1 Chains / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012720:Severity of Illness Index; D013019:South Africa / Q000453:epidemiology; D055815:Young Adult
chemical_list: D000209:Acute-Phase Proteins; D001323:Autoantibodies; D015415:Biomarkers; D016207:Cytokines; D018984:Epitopes, T-Lymphocyte; D059811:HLA-DRB1 Chains