Research to develop instrumentatIon and methods for the sequence analysis of peptide antigens presented to the immune system in association with class I and class II molecules of the major histocompatibility complex is proposed. This is a necessary first step in the development of synthetic/recombinant vaccines or other modulators of the immune system that are effective against bacterial and viral infections, cancer, autoimmune disorders and tissue transplant rejection. Of highest priority is the development of methods for characterizing specific disease state associated antigens found in complex mixtures of several thousand self peptides. Multistage chromatography in conjunction with both microcapillary high performance liquid chromatography/ and high performance capillary electrophoresis/ electrospray ionization tandem mass spectrometry will be employed for this purpose. Both of the above systems will be coupled directly to cytotoxicity assays. Specific goals include the following: (a) to further improve the sensitivity of the tandem mass spectrometry method so as to facilitate sequence analysis of class I peptides at the low femtomole level, (b) to develop proteolytic digestion methods that will facilitate sequence analysis of class II antigens at the low femtomole level, (c) to sequence peptides presented by class I HLA- A2.l molecules on human melanoma cells that are recognized by melanoma specific cytotoxic T lymphocytes, (d) to sequence peptides presented by the human class I molecules, HLA-B7, Aw68, and Aw69, (e) to determine the structural basis of peptide interaction with class I, MHC molecules and to define peptide epitopes recognized by xenoreactive, HLA-A2.1 restricted CTL, (f) to sequence peptides processed from HIV virus proteins and presented in association the class 1, HLA-A2.l molecules, (g) to identify the range of endogenous influenza hemagglutinin pep tides associated with the murine MHC class 1 Kd molecule produced by the wild type hemagglutinin gene and synthetic minigenes and to identify endogenous hemagglutinin peptides that bind Kd but are not immunogenic.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI033993-04
Application #
2003931
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1994-01-01
Project End
1998-12-31
Budget Start
1997-01-01
Budget End
1997-12-31
Support Year
4
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Virginia
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
001910777
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Mohammed, Fiyaz; Stones, Daniel H; Zarling, Angela L et al. (2017) The antigenic identity of human class I MHC phosphopeptides is critically dependent upon phosphorylation status. Oncotarget 8:54160-54172
Sim, Malcolm J W; Malaker, Stacy A; Khan, Ayesha et al. (2017) Canonical and Cross-reactive Binding of NK Cell Inhibitory Receptors to HLA-C Allotypes Is Dictated by Peptides Bound to HLA-C. Front Immunol 8:193
Pavlos, Rebecca; McKinnon, Elizabeth J; Ostrov, David A et al. (2017) Shared peptide binding of HLA Class I and II alleles associate with cutaneous nevirapine hypersensitivity and identify novel risk alleles. Sci Rep 7:8653
Bergmann, Tobias; Lindvall, Mikaela; Moore, Erin et al. (2017) Peptide-binding motifs of two common equine class I MHC molecules in Thoroughbred horses. Immunogenetics 69:351-358
Malaker, Stacy A; Penny, Sarah A; Steadman, Lora G et al. (2017) Identification of Glycopeptides as Posttranslationally Modified Neoantigens in Leukemia. Cancer Immunol Res 5:376-384
Weisbrod, Chad R; Kaiser, Nathan K; Syka, John E P et al. (2017) Front-End Electron Transfer Dissociation Coupled to a 21 Tesla FT-ICR Mass Spectrometer for Intact Protein Sequence Analysis. J Am Soc Mass Spectrom 28:1787-1795
Mohammed, Fiyaz; Stones, Daniel H; Zarling, Angela L et al. (2017) The antigenic identity of human class I MHC phosphopeptides is critically dependent upon phosphorylation status. Oncotarget :
Malaker, Stacy A; Ferracane, Michael J; Depontieu, Florence R et al. (2017) Identification and Characterization of Complex Glycosylated Peptides Presented by the MHC Class II Processing Pathway in Melanoma. J Proteome Res 16:228-237
Sidney, John; Schloss, Jennifer; Moore, Carrie et al. (2016) Characterization of the peptide binding specificity of the HLA class I alleles B*38:01 and B*39:06. Immunogenetics 68:231-6
Zhang, Lichao; English, A Michelle; Bai, Dina L et al. (2016) Analysis of Monoclonal Antibody Sequence and Post-translational Modifications by Time-controlled Proteolysis and Tandem Mass Spectrometry. Mol Cell Proteomics 15:1479-88

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