Association of a 9.2‐kilobase pvu ii class i major histocompatibility complex restriction fragment length polymorphism with ankylosing spondylitis

Abstract

We analyzed DNA restriction fragment length polymorphism (RFLP) in 53 white ankylosing spondylitis (AS) patients and 92 healthy controls, utilizing a full-length HLA-B7 complementary DNA probe. A 9.2-kilobase (kb) Pvu I1 fragment was found to be significantly increasexn frequency in B27 positive AS patients compared with the B27 positive control group (P = 0.00085, relative risk = 7.2). The presence of both B27 and the 9.2-kb RFLP in an individual conferred a relative risk for AS of 297, compared with a relative risk of 119 in those who had B27 alone. The 9.2-kb RFLP appears to be a marker for AS which, with HLA-B27, contributes further to susceptibility to the disease. Our The seronegative spondylarthropathies, including ankylosing spondylitis (AS), Reiter's syndrome, psoriatic arthritis/spondylitis, and the arthritidspondylitis of inflammatory bowel disease, constitute a heterogeneous group of disorders characterized by axial and/or peripheral arthritis, overlapping clinical features, a tendency toward familial aggregation, and the absence of rheumatoid factor or subcutaneous nodules (1). Although the association of AS and other seronegative spondylarthropathies with HLA-B27 is well documented (2,3), the precise role of B27 in the genetic susceptibility to AS is yet to be established. There is a marked discrepancy between the frequency of this antigen in the white population (6.8%) and the prevalence of AS (0.2%), which raises the possibility that a unique subtype of B27 or a linked gene might be more disease-specific. As many as 6 B27 subtypes have been identified by a combination of approaches, including monoclonal antibody studies (4), cytotoxic T lymphocyte studies (5,6), a restriction typing assay (7), high performance liquid chromatography peptide mapping @), and amino acid and nucleotide sequencing (9,lO). Different B27 variants have exhibited higher or lower prevalence according to ethnic group (1 1,12).The availability of complementary DNA (cDNA) probes specific for the class I major histocompatibility complex (MHC) region genes permits further definition of these genes at the DNA level. A useful approach in this regard has been the technique of restriction fragment length polymorphism (RFLP)

Molecular analysis of major histocompatibility complex alleles associated with the lupus anticoagulant.

“…Genomic DNA was extracted from peripheral blood leukocytes (18) and 10 ug digested with the restriction endonucleases Bam HI and Taq I under conditions specified by the manufacturer, and then electrophoresed through 0.8% agarose gel overnight at 50 V. The gel was then stained in ethidium bromide, photographed, and subjected to denaturation and neutralization as described previously (19), and transferred onto Zetabind membranes (Cuno Inc., Meriden, CT).…”

Association of amino acid sequences in the HLA-DQB1 first domain with antitopoisomerase I autoantibody response in scleroderma (progressive systemic sclerosis).

“…"Broad" HLA-DR specificities (DR1, DR2, DR3/6, DR4, DR5(DRwll), DR5(DRwl2), DR7, DRw8, DR9, and DRw1O) were determined either by RFLP analysis of Taq I, Bam HI and Eco RI digested genomic DNA described previously (42) or by polymerase chain reaction (PCR) amplification of 0.5 gg of genomic DNA using the primers DRBAMP-A (5'-CCCCACAGCACGTTTCTTG-3') and DRBAMP-B (5'-CCGCTGCACTGTGAAGCTCT-3'). The PCR was carried out for 30 cycles under the following parameters: denaturation 1 min at 96°C, annealing I min at 55°C, extension 2 min at 72°C.…”

Association of polar amino acids at position 26 of the HLA-DQB1 first domain with the anticentromere autoantibody response in systemic sclerosis (scleroderma).

“…A variety of studies have reported associations of SS with class I MHC specificities, includingHLA-A9 (11,12), B8 (13,14), andB35 (15,16); class II MHC, including HLA-DRl (16)(17)(18)(19), DR3 (14,20) and DR5 (19,(21)(22)(23); and class III MHC, specifically C4 null alleles (15,22). These associations have been generally weak and inconsistent between different centers, and some investigations have even been unable to show any significant HLA associations with SS (24)(25)(26)(27)(28) (33); 10 sg was digested with the restriction endonucleases BamH 1 and TaqI under conditions specified by the manufacturer, and then electrophoresed through an 0.8% agarose gel overnight at 50 V. The gel was then stained in ethidium bromide, photographed, subjected to denaturation and neutralization as described previously (34), and then transferred on to Zetabind membranes. (Cuno, Inc., Meriden, CT).…”

“…Oligonucleotide probes corresponding to the polymorphisms at positions 25,34,55, and 69 were used forthe hybridizations as supplied by the 11th International Histocompatibility Testing Workshop.…”