The overall objective of this project is the development of a simple, rapid, and precise method of typing the polymorphism at the HLA class II (in particular, DRB1, DQB1, DPB1, and DQA1) and the class I (HLA-A, B, and C) loci using the polymerase chain reaction (PCR) method of DNA amplification. Our experimental strategy is to use, when possible, locus specific PCR primers and a panel of sequence specific oligonucleotide probes. Our goal is to develop a simple and convenient """"""""reverse dot blot"""""""" assay for each locus which involves the hybridization of a labeled PCR product to a strip containing a defined array of immobilized probes in a single reaction, as well as an automated approach to """"""""reading"""""""" the genotype from the pattern of probe reactivity. This project will involve the sequence analysis of class I and class II genes from non-Caucasian populations and families to identify new alleles and haplotypes as well as the determination of allele, phenotype, and genotype frequencies in various human populations using oligonucleotide typing. Unlike immunological approaches or RFLP analysis, methods for detecting coding sequence polymorphisin, such as oligonucleotide typing, are valuable because they indicate where and how alleles differ. Individual residues are known to elicit allogeneic T cell responses; consequently, this typing approach should prove valuable for transplantation. In particular, it should help us determine the nature and location of mismatches which are important in determining the clinical outcome of bone marrow transplants. In addition, these assays will continue to help us identify specific class II polymorphic residues which may be implicated in disease susceptibility. They should also prove valuable for studies of peptide, MHC, and T cell receptor interactions.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL047170-01
Application #
3366402
Study Section
Special Emphasis Panel (SRC (MR))
Project Start
1991-08-01
Project End
1995-05-31
Budget Start
1991-08-01
Budget End
1992-05-31
Support Year
1
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Roche Molecular Systems, Inc.
Department
Type
DUNS #
City
Alameda
State
CA
Country
United States
Zip Code
94501
Spencer, Bryan R; Johnson, Bryce; Wright, David J et al. (2016) Potential impact on blood availability and donor iron status of changes to donor hemoglobin cutoff and interdonation intervals. Transfusion 56:1994-2004
Spencer, Bryan R; Kleinman, Steven; Wright, David J et al. (2013) Restless legs syndrome, pica, and iron status in blood donors. Transfusion 53:1645-52
Carrick, Danielle M; Johnson, Bryce; Kleinman, Steven H et al. (2011) Agreement among HLA antibody detection assays is higher in ever-pregnant donors and improved using a consensus cutoff. Transfusion 51:1105-16
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Shaz, Beth H; Demmons, Derrick G; Crittenden, Colleen P et al. (2009) Motivators and barriers to blood donation in African American college students. Transfus Apher Sci 41:191-7
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Trachtenberg, E A; Keyeux, G; Bernal, J E et al. (1996) Results of Expedicion Humana. I. Analysis of HLA class II (DRB1-DQA1-DPB1) alleles and DR-DQ haplotypes in nine Amerindian populations from Colombia. Tissue Antigens 48:174-81
Trachtenberg, E A; Erlich, H A; Rickards, O et al. (1995) HLA class II linkage disequilibrium and haplotype evolution in the Cayapa Indians of Ecuador. Am J Hum Genet 57:415-24

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