HLA‐DQB1 allele typing by a new PCR‐RFLP method: Correlation with a PCR‐SSO method

Salazar, Marcela; Yunis, Juan J.; Delgado, Maria Belen; Bing, David H.; Yunis, Edmond J.

doi:10.1111/j.1399-0039.1992.tb02102.x

Cited by 34 publications

(21 citation statements)

References 13 publications

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“…The primers and conditions used in this study as well as dot blots, prehybridization and hybridization procedures were as previously described [22,231. The DRB, DQAl and DQBl alleles were determined using the locusspecific PCR amplified products by PCR-SSO as described before [22, 231.…”

Section: Hla Polymorphism Proceduresmentioning

confidence: 99%

Polymorphic human specific Alu insertions as markers for human identification

Novick

Yunis

et al. 1995

Electrophoresis

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Alu sequences represent the largest family of short interspersed repetitive elements (SINEs) in humans with 500 000 copies per genome. Recently, one Alu subfamily was found to be human specific (HS). We originally described the use of polymorphis HS Alu insertions as a tool in population studies and recently as tools in DNA fingerprinting and forensic analysis. In this report, we will use this simple polymerase chain reaction (PCR) base technique for the detection of HS Alu insertion polymorphisms. We will test the resolving power of this DNA profiling approach in both population genetics and paternity assessment. At the population level, we will describe the genotypic distribution of five polymorphic Alu insertions among 3 populations from the American continent, one of African origin, the other two Amerindians. Insight into their relationships will be provided. At the family level, we will examine one European American family of seven individuals and the same pedigree will also be characterized by way of the two systems currently and widely used to ascertain paternity: PCR-sequence specific oligonucleotide probe hybridization (PCR-SSO) and PCR-restriction fragment length polymorphism (PCR-RFLP) of human leucocyte antigen (HLA) class II molecules, and a standard RFLP protocol used in forensic casework and paternity studies. The importance and strengths of the methods as well as its perspectives for future use in filiation studies will be evaluated.

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Section: Hla Polymorphism Proceduresmentioning

confidence: 99%

Polymorphic human specific Alu insertions as markers for human identification

Novick

Yunis

et al. 1995

Electrophoresis

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“…11,12 TNFb (tumor necrosis factor microsatellite b) microsatellite variants were determined as described. 13 HLA typing of the patient was achieved using the technique of sequence-specific primers (SSP) as previously described.…”

Section: Detection Of Donor Cell Engraftmentmentioning

confidence: 99%

Transplantation of unrelated cord blood cells

Weinreb

Delgado

Clavijo

et al. 1998

Bone Marrow Transplant

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Summary:A 43-year-old woman with Philadelphia chromosome (Ph) positive chronic myelogenous leukemia in acute phase received high-dose chemotherapy followed by transfusion of 12 randomly selected units of umbilical cord blood. HLA analysis showed cells of one donor from day +10 to day +43 post-transfusion. This unit was HLA class II identical with that of the patient. Keywords: placental blood; unmatched; multiple donors; CML There is considerable interest in umbilical cord blood (UCB) transplants (for reviews see Refs 1-4). Most transplants have been from HLA-matched relatives. However, there is increasing use of HLA-matched unrelated donors (reviewed in Ref.3). In 1972, Ende et al 5 transfused UCB from eight non-HLA unrelated donors to a 16-year-old with ALL previously treated with conventional chemotherapy. Success was claimed based on showing RBCs from one donor for 8 months. Shen et al 6 described giving multiple units of HLA-unmatched UCB to four patients with solid tumors. A patient with CML in accelerated phase was transplanted with HLA-partially matched UCB from one donor by Laporte et al 7 who reported successful transplantation of the cord blood stem cells into a 26-year-old women. We report a patient who received 12 units of HLA-untyped UCB for CML in acute phase after high-dose chemotherapy. Case reportThe patient, a 43-year-old female, was diagnosed with Ph chromosome-positive CML in 1991. She was treated with hydroxyurea and interferon until 1993 when accelerated phase developed. She then received total body irradiation (TBI) and cyclophosphamide followed by an autotransplant. In April 1996 accelerated phase recurred. She was treated with cytarabine to which she responded briefly. Based on a previously approved Institutional Review Board (IRB) transplantation protocol involving unrelated UCB and after informed consent, the patient received busulfan (12 mg/kg) and cyclophosphamide (120 mg/kg) followed by infusion of 3.18 × 10 9 mononuclear cells obtained from UCB from 12 unrelated donors.On day +1 we began GVHD prophylaxis consisting of cyclosporine and solumedrol. Early signs of engraftment were observed on day +10 with islands of trilineage hematopoiesis noted in a bone marrow biopsy specimen. There were no clinical features of GVHD. Increased bilirubin developed between days +14 and +17 with a peak value at 7.6 mg/dl. The patient was discharged in a stable condition on day +21 with a WBC count of 2.6 × 10 9 /l. Fetal hemoglobinized RBC and RhD-positive RBC were detected in the blood between day +17 and day +21 (patient's blood type O RhD-negative). Southern blot analysis of DNA from blood cells on day +16 showed no bcr/abl rearrangement (previously detected). Growth factor therapy was stopped on day +31 when the WBC count was Ͼ10 × 10 9 /l. The WBC continued to rise and was 8.5 × 10 9 /l on day +53 with reappearance of Ph chromosome-positive cells. Hydroxyurea and interferon were begun on day +71 and she died on day +90 of leukemia. Umbilical cord blood collecting and processingUnits of UCB were ...

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“…Dot-blot, prehybridization, and hybridization procedures were carried out as described (12, 13). DRB1 and DQB1 alleles were determined in the PCRamplified products by sequence-specific oligonucleotide probe hybridization as described before (11,12).Statistical Analysis. The frequencies of independent haplotypes and alleles were determined by direct counting.…”

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confidence: 99%

“…DNA samples were amplified by PCR for DRB1 and DQB1 loci in a 100-,ul reaction mixture containing 50 mM KCl, 10 mM Tris HCl (pH 8.3), dNTPS (each at 125 mM), 50 pmol of each primer, 1.5-2 mM of MgCl2, and 2.5 units of Taq polymerase [AmpliTaq DNA polymerase (Perkin-Elmer/Cetus)]. The primers and conditions used in this study have been described elsewhere (10)(11)(12)(13)(14). Negative controls were included to detect contamination.…”

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confidence: 99%

A common major histocompatibility complex class II allele HLA-DQB1* 0301 is present in clinical variants of pemphigoid.

Delgado

Turbay

Yunis

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

191

117

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Bullous pemphigoid (BP) is an autoimmune subepidermal blistering disease seen primarily in elderly persons. It is characterized clinically by the development of tense bullae and by the presence of an antibasement membrane antibody. In BP, the antigens involved in the autoimmunity are epidermal basement membrane peptides BPAgl and BPAg2. We have compared high resolution typing of major histocompatibility complex class II loci (HLA-DRB1, DQB1) in 21 patients with BP, 17 with ocular cicatricial pemphigoid (OCP), and 22 with oral pemphigoid (OP) to a panel of 218 haplotypes of normal individuals. We found that the three diseases (BP, OCP, and OP) have significant association with DQB1*0301 (P = 0.005, P < 0.0001, and P = 0.001, respectively). The frequencies of alleles DQB1*0302, *0303, and *06, which share a specific amino acid sequence from position 71 to 77 (Thr-Arg-Ala-Glu-Leu-Val-Thr) were also increased (P = 0.01). We suggest that an identical major histocompatibility complex class II allele (DQB1*0301) is a common marker for enhanced susceptibility and that the same amino acid residues in positions 71-77 (DQB1*0301, -0302, -0305, -0602, -0603 alleles) are found in patients with BP, OCP, and OP. Our findings propose that the autoimmune response in the three different clinical variants of pemphigoid, involves the recognition by T cells of a class II region of DQB1, bound to a peptide from the basement membrane of conjunctiva, oral mucosa, and skin.Bullous pemphigoid (BP) is an autoimmune subepidermal blistering disease seen primarily in elderly persons and is characterized clinically by the development of blisters. Therapy usually involves oral corticoids and frequently immunosuppressant agents (1). There is in vivo deposition of immunoglobulins (Ig) and complement components at the basement membrane zone of the dermo-epidermal junction. Sera of the majority of patients contains anti-basement membrane zone autoantibodies (1).It is postulated that the lesions in BP result from autoimmunization with IgG class autoantibodies against antigens BPAgl and BPAg2, components of the epidermal basement membrane zone (2). Recently, it has been demonstrated that passive transfer of autoantibodies that recognize BPAg2, when injected into neonatal mice, can produce clinical BP (3).In previous studies we have shown an association between the DQB1*0301 allele with ocular cicatricial pemphigoid (OCP) and oral pemphigoid (OP), when compared with a population of normal controls (4, 5). While in patients with pemphigus vulgaris we have found an association with HLA-DR4, DQ8 among Jewish patients, and DR6, DQ5 in nonJewish patients (6-8).In the present report, 21 Caucasian patients with BP were studied for major histocompatibility complex (MHC) class II DRB1, DQB1 alleles, using amplification of genomic DNA by the polymerase chain reaction and sequence-specific oligonucleotide probe hybridization. Frequencies were compared with 218 haplotypes of normal individuals. MATERIALS AND METHODSTwenty-one patients with BP were typed ...

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HLA‐DQB1 allele typing by a new PCR‐RFLP method: Correlation with a PCR‐SSO method

Cited by 34 publications

References 13 publications

Polymorphic human specific Alu insertions as markers for human identification

Polymorphic human specific Alu insertions as markers for human identification

Transplantation of unrelated cord blood cells

A common major histocompatibility complex class II allele HLA-DQB1* 0301 is present in clinical variants of pemphigoid.

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