The success or failure of tissue allografts is determined by the products of a large family of autosomal and sex-linked, polymorphic genes that are designated histocompatibility (H) genes. The strongest barriers to successful transplantation are determined by the alloantigenic products of class I and class II genes of the major histocompatibility complex (MHC). The matching of donors and recipients for MHC class I and class II alloantigens not only reduces or eliminates MHC-encoded barriers but, conversely, optimizes the presentation of donor minor H antigens that require MHC matching of donor and recipient for presentation. The demonstration that the antigenic components of single minor H antigens are peptides bound to the peptide binding site of MHC molecules provides not only a reason for why minor H genes have eluded cloning, but, also provides an experimental approach to identifying and sequencing these H antigen peptides. Using the knowledge of the class I molecule that is required for the presentation of a single minor H antigen to cytolytic T lymphocytes (CTL) and the fact that subsets of endogenous peptides bind to individual class I molecules, we have chromatographically resolved peptides from multiple minor H antigens. The amino acid sequences of these minor H peptides can be applied to cloning and sequencing the respective genes. The overall objectives of this research program are to clone and identify minor H genes in the mouse and to understand the specificity and genetic control of the T cell response to their encoded antigens.
The first aim i s the amino acid sequencing of minor H antigen peptides by mass spectrometry and application of the amino acid sequences to the synthesis of degenerate oligonucleotides for the cloning of minor H gene CDNAS. H gene CDNAS will be confirmed by sequencing and DNA-mediated gene transfer into eukaryotic cells with subsequent testing with specific CTL. The second objective is the estimation of diversity of minor H peptides expressed by inbred mouse strains by amplification and sequencing of DNA sequences that encode the homologous peptides. The third objective is the testing of our hypothesis that minor H peptides comprise a group of non-abundant endogenous peptides that exhibit relatively higher affinity for class I molecules than that exhibited by abundant, endogenous peptides. Peptides derived from minor H antigens, viral and soluble antigens, and abundant endogenous proteins will be tested for relative affinity in competitive binding assays and assays for stabilization of expression of class I:peptide complexes. The fourth objective is the sequencing of immunodominant minor H antigen peptides and the testing of the hypothesis that the ranking of these dominant antigens is based on the relative affinity of their peptides for class I molecules. These studies constitute the first analysis of the T cell response to minor H antigens that will both sequence minor H antigen peptides and genes as well as investigate the affinity and specificity of the binding of these peptides to MHC class I molecules.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI016052-18
Application #
2060325
Study Section
Immunological Sciences Study Section (IMS)
Project Start
1989-12-01
Project End
1998-07-31
Budget Start
1994-08-01
Budget End
1995-07-31
Support Year
18
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Mayo Clinic, Rochester
Department
Type
DUNS #
City
Rochester
State
MN
Country
United States
Zip Code
55905
Wettstein, Peter J; Borson, Nancy D (2007) Distributions of single nucleotide polymorphisms in differential chromosome segments of congenic resistant strains that define minor histocompatibility antigens. Immunogenetics 59:631-9
Wettstein, Peter J; Strausbauch, Michael; Borson, Nancy (2007) Repertoires of T cell receptors expressed by graft-infiltrating T cells evolve during long-term recall responses to single minor histocompatibility antigens. Int Immunol 19:523-34
Wettstein, Peter J; Borson, Nancy D; Park, Jewn G et al. (2005) Cysteine-tailed class I-binding peptides bind to CpG adjuvant and enhance primary CTL responses. J Immunol 175:3681-9
Lanza, Robert P; Chung, Ho Yun; Yoo, James J et al. (2002) Generation of histocompatible tissues using nuclear transplantation. Nat Biotechnol 20:689-96
Nevala, W K; Paul, C; Wettstein, P J (1998) Immunodominant minor histocompatibility antigen peptides recognized by cytolytic T lymphocytes primed by indirect presentation. Transplantation 65:559-69
Strausbauch, M A; Nevala, W K; Roopenian, D C et al. (1998) Identification of mimotopes for the H4 minor histocompatibility antigen. Int Immunol 10:421-34
Borson, N D; Strausbauch, M A; Wettstein, P J et al. (1998) Direct quantitation of RNA transcripts by competitive single-tube RT-PCR and capillary electrophoresis. Biotechniques 25:130-7
Johnston, S L; Graham, D; Wettstein, P J (1997) Diversity of alpha and beta subunits of T-cell receptors specific for the H4 minor histocompatibility antigen. Immunogenetics 46:17-28
Nevala, W K; Wettstein, P J (1997) Immunodominant minor histocompatibility antigen peptides presented by H2Db molecules. Transplantation 64:1323-30
Nevala, W K; Paul, C; Wettstein, P J (1997) Reduced diversity of CTLs specific for multiple minor histocompatibility antigens relative to allograft rejection in vivo. J Immunol 158:1102-7

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