Annual and Interim Progress Report Summaries
Principal Investigator: Richard Koup
Project: Comprehensive T Cell Vaccine Immune Monitoring Consortium
Submitted January 20, 2010 (Interim Report)
Year 4 activities for the CTC-VIMC include:
• Supporting the Ho, EuroVac, DC VAC001 and Shattock clinical trials and the Picker (Parks IAVI VDC) NHP trial;
• Qualifying new ICS assays for clinical (7 color) and NHP (8 color) laboratories;
• Quarterly performance monitoring of the standardized IFNγ ELISpot assay in our 4 clinical core labs and harmonizing use of the BIDMC ELISpot across 3 NHP labs; and
• Ramping up the new Molecular Immune Correlates Discovery (MICD) initiative.
Thus, our year 4 agenda balances T cell immunological CAVD infrastructure refinement with an innovative discovery initiative.
A new MICD workgroup comprised of members representing our PBMC Repository (Proficiency Testing Core), NHP Scientific Core and two of our discovery teams (Immune Correlates and the Innate and Mucosal Immunity), will work collaboratively to bring new technology to bear on efforts to further characterize antigen-specific T cell responses. This MICD group will use epitope mapping, tetramer staining and TCR clonotyping to evaluate the breadth and clonality of immune responses in cohorts of clinical and NHP specimens. Goals for this year include characterizing responses in CMV+, HIV- and HIV+ (acute and LTNP) human subjects (including B*57 and A*022 responders) using PBMC and gut biopsies. NHP specimens will include MamuA01 animals that have received DNA/Ad vaccine and others that have received the CMV vaccine. These same specimens will then undergo microarray analysis to determine gene specific responses, which will then be validated through the use of the Fluidigm technology. Use of Fluidigm single cell technology will provide additional information on dynamic range and the heterogeneity of expression.
Efforts in the first half of year 4 have focused on refining the tetramer sorting and staining techniques, epitope mapping repository specimens, and establishing the availability of other specimens for analysis. Currently, we are finalizing a 3 hour tetramer staining and sorting procedure capable of distinguishing CD4 responses, as well as the originally planned CD8 response. Once finalized, cell sorting will begin, followed by clonotyping, microarrays and the Fluidigm analyses. Planning for expansion of this project in year 5, we continue to build alliances with a variety of vaccine development groups to enable testing a broad variety of vaccine responses.