The long term objective of this application is to elucidate pathways for generating an MHC molecule/peptide complex within an antigen presenting cell.
The specific aims of this proposal are to functionally and genetically characterize a group of mutant antigen presenting cells, which are unable to present a variety of soluble antigens to class II restricted T cells but can present a peptide antigen. The approaches are: to use proteolytic digests of antigen and DR restricted peptide specific T cell clones to more extensively evaluate the mutants' ability to present antigen; to determine whether the mutants are defective in antigen processing via the endocytic pathway; to use somatic cell hybrids to determine how many complementation groups exist among 8 obligate independent presentation defective mutants; to determine whether human non-lymphoid cells and murine lymphoid cells can complement the defect(s) in the mutants; to determine whether the affected gene(s) map to X chromosome or the HLA-D region of chromosome 6; and to isolate phenotypically similar mutants using retroviral insertional mutagenesis as a tool for eventual cloning of the mutant gene(s). In addition, isolation of other antigen processing mutants by immunoselection with antigen specific cytolytic T cells will be pursued. These studies may lead to a better understanding of MHC restricted antigen processing/presentation and may thus provide insights into the pathophysiology of HLA associated diseases in which MHC restricted T cells are likely to play a role.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29AI028809-04
Application #
3455349
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1989-12-01
Project End
1994-11-30
Budget Start
1992-12-01
Budget End
1993-11-30
Support Year
4
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Children's Hospital of Philadelphia
Department
Type
DUNS #
073757627
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Hou, Tieying; Rinderknecht, Cornelia H; Hadjinicolaou, Andreas V et al. (2013) Pulse-chase analysis for studies of MHC class II biosynthesis, maturation, and peptide loading. Methods Mol Biol 960:411-432