DNA sequences that characterize the class I and class II HLA alleles can be identified by hybridization with oligonucleotide probes. In the present studies we will use selective amplification of genomic DNA by the polymerase chain reaction (PCR) followed by hybridization with sequence-specific probes. Locus-specific and group-specific PCR will be used. The latter will be preferred when otherwise correct assignment would be difficult, due to the sharing of polymorphic sequences between alleles. Because of the large number of polymorphisms and the complex sharing of sequences, typing methods will be developed by groups of related alleles. Radioactive and non-radioactive batch-mode procedures will be used, and a method oriented to typing of single samples, using an enzyme-linked immunoassay with magnetic beads, will be explored. The latter approach will make use of 96-well trays pre-loaded with oligonucleotide probes and will be suitable for automation of the typing procedure. Population studies will include North American Caucasoids, American Blacks, Mexican-Americans, and North American Indians. Variant alleles prevalent in each of these populations will be analyzed. it is anticipated that some new alleles will be described on the basis of hybridization patterns. They will be confirmed by sequencing. The functional role of the polymorphic sequences determined by oligonucleotide hybridization, will be investigated using primary mixed lymphocyte cultures (MLC) and T cell clones. In these experiments we will attempt to determine the correlations between polymorphic residues and T cell recognition. Detailed information to be developed will be useful to understand the immune response, HLA associated diseases and the impact of HLA polymorphisms in clinical transplantation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL047145-02
Application #
3366360
Study Section
Special Emphasis Panel (SRC (MR))
Project Start
1991-08-01
Project End
1995-05-31
Budget Start
1992-06-01
Budget End
1993-05-31
Support Year
2
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Type
Schools of Medicine
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390
Lin, M S; Fu, C L; Aoki, V et al. (2000) Desmoglein-1-specific T lymphocytes from patients with endemic pemphigus foliaceus (fogo selvagem). J Clin Invest 105:207-13
Zwirner, N W; Dole, K; Stastny, P (1999) Differential surface expression of MICA by endothelial cells, fibroblasts, keratinocytes, and monocytes. Hum Immunol 60:323-30
Forero, L; Zwirner, N W; Fink, C W et al. (1998) Juvenile arthritis, HLA-A2 and binding of DEK oncogene-peptides. Hum Immunol 59:443-50
Zwirner, N W; Fernandez-Vina, M A; Stastny, P (1998) MICA, a new polymorphic HLA-related antigen, is expressed mainly by keratinocytes, endothelial cells, and monocytes. Immunogenetics 47:139-48
Marcos, C Y; Fernandez-Vina, M A; Lazaro, A M et al. (1997) Novel HLA-B35 subtypes: putative gene conversion events with donor sequences from alleles common in native Americans (HLA-B*4002 or B*4801). Hum Immunol 53:148-55
Fernandez-Vina, M A; Lazaro, A M; Marcos, C Y et al. (1997) Dissimilar evolution of B-locus versus A-locus and class II loci of the HLA region in South American Indian tribes. Tissue Antigens 50:233-50
Wang, J; Fernandez-Vina, M A; Lazaro, A M et al. (1997) New alleles of the HLA-B15 family. Hum Immunol 55:184-9
Rani, R; Fernandez-Vina, M A; Zhang, S et al. (1995) HLA-DPB1 alleles in a population from north India and description of a new variant (DPB1*5601). Tissue Antigens 45:264-9
Satz, M L; Fernandez-Vina, M; Theiler, G C et al. (1995) Allelic heterogeneity of HLA-B35 subtypes in different populations as assessed by DNA typing. Tissue Antigens 46:196-203
Fernandez-Vina, M; Lazaro, A M; Sun, Y et al. (1995) Population diversity of B-locus alleles observed by high-resolution DNA typing. Tissue Antigens 45:153-68

Showing the most recent 10 out of 23 publications