Koup T-cell Vaccine Immune Monitoring Consortium

OVERVIEW:

The goal of the Comprehensive T-cell Vaccine Immune Monitoring Consortium (T-cell VIMC) is to create a consortium of clinical and Non-Human Primate (NHP) immune testing laboratories within the top HIV vaccine pre-clinical and clinical trials networks. This will provide the ability to standardize certain laboratory practices and assays across clinical trials and to bring together leading vaccine scientists to identify new targets and assays for use in T-cell immune monitoring. The T-cell VIMC will enable researchers to optimize and standardize the best assays for general distribution and use in comparing different vaccine platforms. The end product will be a system that allows for the intelligent comparison of T-cell immunogenicity of pre-clinical and clinical HIV vaccine platforms within the context of an ever-expanding knowledge base on HIV transmission and basic principles of immunology. 

RESEARCH OBJECTIVES:

1. Establish an international consortium of four GCLP clinical laboratories and three central NHP laboratories with centralized coordination, standardization, and certification of T-cell immune testing.

2. Establish discovery teams to develop new assays and technologies for monitoring T-cell immune responses to HIV vaccines.

3. Provide administrative and scientific structure that will facilitate the evaluation of new assays developed by the discovery teams leading to rapid validation, reagent standardization, training, and distribution to central laboratories.

PROGRESS:

With both clinical and NHP core capacity established, standardization of ELISpot assay achieved, and development of expanded intracellular cytokine staining (ICS) panels well in hand, current efforts focus on bringing innovative technologies to bear on the search for correlates of protection. Building on recent efforts to comparatively evaluate alternative T-cell research assays such as the cultured ELISpot, CFSE and viral inhibition assays, the CTC-VIMC will now focus its discovery efforts on new methods to characterize antigen specific T-cell responses, including breadth of responses through epitope mapping, the clonality of responses through TCR clonotyping, and gene specific responses through microarrays which will then be validated through an innovative single cell technology to provide additional information on dynamic range and the heterogeneity of expression. To achieve the broadest applicability of results, these new methods will be deployed with specimens derived from clinical and NHP trials across a variety of vaccine candidates.

Specific achievements include:

• Provided T cell assay services to clinical trials including ADVAX-Electroporation (C004) and EV03. Support of DC-VAX001 to begin in 2010.
• Established Reagent Core with ongoing distribution of peptides to CAVD researchers (e.g., SIVmac239, SIVmacE543, Ad5, HIV, CMV, CEF.  PTE peptides coming in 2010.)
• Optimized efficient epitope-mapping strategy that uses 4 stage mapping procedure with a two dimensional matrix.
• Conducted comparative evaluation of T cell research assays including cultured ELISpot, CFSE, Viral Inhibition Assay, and perforin ICS assay.
• Established clinical and NHP PBMC banks from leukaphresis programs at Duke Human Vaccine Institute and the VRC, respectively. Clinical specimens include fully-characterized and HLA-typed HIV+ and HIV- donors.  NHP PBMCs include CITES permitted animals with exposures to various HIV vaccine regimens. Currently augmenting clinical blank with HIV+ (ART naïve) and HIV- specimens from Africa.  Specimens used for assay development and proficiency testing initiatives.
• Determined utility of PTE peptides for cross-trial comparisons.
• Completing qualification of 7 color clinical ICS panel that includes markers for CD3, CD4, CD8, INFγ, IL-2 and TNFα and viability; parallel development of 8 color panel for NHP laboratories at pre-qualification stage. Assay development testing included collaborative testing at 8 clinical laboratories and 5 NHP laboratories
• Four Clinical Core laboratories, representing the HVTN, IAVI and the VRC’s NVITAL, established equivalency of their respective ELISpot assays through rigorous comparative statistical evaluation and technology transfer. Maintenance of equivalency to be monitored by quarterly proficiency testing.  Parallel testing program ongoing in NHP laboratories.
• Four Core Clinical Laboratories GCLP certified.
• Evaluating cohorts of LTNP and YF vaccinees to identify new T cell functions that correlate with protection from disease and progression.
• Established genomic profiles relevant to several vaccine platforms and trials; improved reporting procedures for data generated from micro array analyses.
• Optimized Cultured ELISpot assay.