Annual and Interim Progress Report Summaries
Principal Investigator: Richard Koup
Project: Comprehensive T Cell Vaccine Immune Monitoring Consortium
Submitted June 29, 2012
The CTVIMC has successfully transitioned to its new streamlined organization for the next five years of CAVD operation. Our laboratories are now located within 6 institutions—the VRC, BIDMC, Duke, FHCRC, Imperial College and VGTI--and focus primarily on providing cellular assay support services for clinical trials and NHP studies. The administrative core is located at FNIH, with responsibility for coordinating collaboration and communications across the consortium and with other CAVD members, as well as overall management of the CTVIMC project. Our discovery research initiatives now represent a smaller portion of the consortium’s effort, focused on interrogation of biological processes as measured by innovative analytic methods.
CTVIMC investigators produced 10 publications this year. Several documented our ongoing assay standardization activities, highlighting qualification of improved ICS panels for clinical and NHP uses, and optimization of the Fluidigm technology. We achieved a truly remarkable concordance in a head to head comparison of the new VIMC ICS panels against the validated FHCRC/HVTN panel. The highly multiplexed Fluidigm technology allows rapid and economical screening of a great many markers potentially related to an immune process from a small or even single cell sample. Through application of the Fluidigm technology to the RV144 specimens, we identified novel markers that could be potential correlates of protection. Other publications documented our findings that: vaccine induced viral inhibition by effector memory CD8+ T cells correlates with better viral control in NHP not protected against infection with SIVsm E660; different Ad serotypes differ in receptor usage, tropism and ability to activate cells and differentially induce production of IFN-alpha and stimulate IFN-related intracellular pathways; and, several innate immune pathways, including interferon and inflammasome signaling, were induced by the adjuvant poly ICLC in a manner similar to that observed with the licensed Yellow Fever vaccine.
In addition to supporting the DCVAX trial and preparing for the MUCOVAC2 trial, investigators within our Clinical Trials Testing Core focused efforts over the year on the optimization of a B cell ELISpot assay, evaluating R848 and CPG-C as alternative stimulations. We have found that stimulation via TLR7 or TLR9 agonists +IL2 induces differentiation and proliferation of B cells into detectable CD27+, CD38+ and CD27/CD38+ IgG producing B cells. In our discovery projects, work to evaluate the mechanism of the Viral Inhibition Assay (VIA) using a luciferase read-out has resulted in significant improvements in the assay including reductions in assay time, background inhibition and cost with a substantial improvement in assay throughput. Our projects interrogating TCR recognition of HIV infected cells and Tfh cells have already developed single cell TCR sequencing techniques and reporter cell lines, and identified that accumulation of Tfh during SIV infection is associated with more GC B cells with high Bcl-6 and higher titers of SIV-specific antibodies.
Other achievements over the past year include: the T cell assay support services our NHP core has provided to several NHP NYVAC trials; fulfillment of orders for HIV and SIV peptides by our Reagent Core, as well as the acquisition of a supply of avi-tagged proteins for use in B cell assay development efforts; and the implementation of an online ordering system by our Central PBMC Repository to facilitate sharing these well characterized specimens with the larger CAVD community.