Identification of human T cell epitopes of pathogenic fungi

The goal of this project is to identify T-cell epitopes of pathogenic fungi including species in the



genera of Blastomyces, Coccidioides, Cryptococcus, and Histoplama. The subcontract site at



UTSA will conduct the proposed experiments for the identification and confirmation of



Coccidioides T cell epitopes. The newly identified T-cell epitopes can be used in an array of



basic immunological research to better understand the fungus-host interaction as well as



translational applications including development of vaccines and diagnostic reagents for



Coccidioides infection. Dr. Hung will direct all experiments at UTSA and collaborate with Dr. Stu



Levitz and other collaborators on this project to complete the following tasks:



1. Compile a set of fast track proteins that have been previously identified and evaluated as



Coccidioides antigens for vaccine and serological diagnosis.



2. Assist in the down selection of Coccidioides antigens and potential T-cell epitopes that



are predicted by bioinformatics analysis.



3. Conduct T-cell activation assays to evaluate immunoreactivity of the predicted T-cell



epitopes from the newly predicted epitopes using human leukocyte antigen (HLA-DR4)



transgenic mice that are vaccinated with the fast track or newly identified Coccidioides



antigens.



4. Hung’s laboratory will prepare samples for evaluation CD4+ T cells response of the



vaccinated and control mice. Samples will be shipped to UMMS for cytokine assays



including Th1 and Th17 responses will be used to delineate the protective immunity



elicited by the vaccine.



5. Hung’s laboratory will prepare human peripheral blood monocytic cells (PBMCs) from



blood samples donated by patients who have recovered from pulmonary



coccidioidomycosis. Dr. Neil Ampel will collect those blood samples and ship them to



Hung’s laboratory. Each predicted epitopes will be incubated with human PBMCs



separately to stimulate T-cell response. The culture supernatants will be collected for



cytokine assays.



6. Hung will communicate with Dr. Stuart Levitz and other collaborators in this project.



Hung’s laboratory will also assist in analyzing data and writing scientific reports.



7. The newly identified and confirmed human T-cell epitopes of Coccidioides will be



deposited into IEDB epitope databased for sharing with the research community.