The Nef allele encoded by the simian immunodeficiency virus (SIV) strain smmPBj14 contains an immunoreceptor tyrosine-based activation motif (ITAM) that interacts with zAP-70 and Syk, and activates intracellular signalling. It is hypothesized that SIVsmmPBj14 Nef may also be capable of stimulating dendritic cells (DC), and that this may enhance antigen presentation by DC.
In aim 1, recombinant adenovirus vectors (rAV) will be used to express the following proteins in DC: (1) wild-type SIVsmmPBj14 Nef, (2) a Y17R mutant, which lacks the ITAM, and (3, 4) truncated mutants which contain only amino acids 1-43 of either wild type or YR mutant Nef (this region includes the intact ITAM). After infection of DC with these vectors, intracellular signaling events and modification of key protein substrates will be examined.
In aim 2, Nef-expressing rAV will be used to infect cultured DC, and the efficiency with which these cells can stimulate an antigen-specific T cell response will be measured. Finally, an rAV construct which co-expresses HIV-1 gp120 and SIVsmmPBj14 Nef (or its ITAM) will be constructed, and inoculated into BALB/c mice; T cell responses to gp120 will be measured. This will reveal whether SIVsmmPBj14 Nef, or its ITAM, can enhance the immunogenicity of an HIV-1 vaccine.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31AI010536-03
Application #
6533980
Study Section
Special Emphasis Panel (ZRG1-RPHB-3 (02))
Program Officer
Hernandez, Milton J
Project Start
2002-09-01
Project End
Budget Start
2002-09-01
Budget End
2003-08-31
Support Year
3
Fiscal Year
2002
Total Cost
$36,625
Indirect Cost
Name
University of Rochester
Department
Microbiology/Immun/Virology
Type
Schools of Dentistry
DUNS #
208469486
City
Rochester
State
NY
Country
United States
Zip Code
14627
Webster, Ramothea L; Johnson, R Paul (2005) Delineation of multiple subpopulations of natural killer cells in rhesus macaques. Immunology 115:206-14