Koup: T Cell Vaccine Immune Monitoring Consortium

OVERVIEW

The goal of the Comprehensive T Cell Vaccine Immune Monitoring Consortium (CTVIMC) is to implement a comprehensive program for assessing vaccine-elicited T cell responses that will facilitate the development and licensure of an HIV vaccine. The consortium provides qualified, GCLP-compliant, standardized T-cell assays and exploratory cellular assays in support of pre-clinical and clinical development of HIV vaccines within the CAVD.​

RESEARCH OBJECTIVES

1. Develop new assays and technologies for monitoring T cell immune responses to HIV vaccines.

2. Maintain a core of central clinical laboratories including the FHCRC/HVTN laboratory, NVITAL and the IAVI HIL at Imperial College.

3. Maintain a core of central NHP laboratories including one at BIDMC and another at the VRC.

4. Enable efficient deployment of T cell assays, oversee new assay transfers and qualifications, and oversee T cell and B cell ELISpot testing for clinical trials and NHP studies within the CAVD.

5. Supply the CAVD community with quality-controlled, characterized PBMC and peptides for use in the assay development and standardization.

6. Provide gene arrays and Fluidigm assays to interrogate vaccine-induced T and B cells for NHP and human clinical trials.

PROGRESS

The CTVIMC continues to provide both standardized T cell assays and a variety of research assays—including an expanding repertoire of B cell assays—to CAVD investigators for both clinical trials and NHP studies. Our preference is to begin working with VDC investigators early in the study design process so that we can optimize assay selection, assuring the best complement of assays and time points for the study, as well as to promote suitable preparation of specimens for cellular assays, and timely procurement of necessary peptides. Our

partnership with each VDC continues throughout the study, convening interim calls as necessary, and lending our collective expertise to investigators as they interpret study results.

Intracellular cytokine staining (ICS) panels remain our primary standardized T cell endpoint assays, with several panels available to measure both CD8 and CD4 responses and a variety of other markers. A standardized IFN-g ELISpot assay is also available from our core of clinical and NHP labs.

Our research assays which serve as secondary endpoints, typically undertaken after a response has been demonstrated in a standardized assay, include:

· ​Cytokine bead array (CBA) to measure factors linked to variety of immune functions, such as Th1, Th2, pro-inflamatory, regulatory, and chemotactic responses;

· ​Epitope mapping, using either ELISpot or ICS platforms to define which epitopes are targeted by a T cell response;

· ​T cell, B cell and NK phenotyping using multi-parameter flow cytometry to define representation and activation state of lymphocyte subsets;

​· ​Fluidigm assays for T and B cell interrogation, both nanoarray and single cell, to quantify gene  expression profiles;

· ​Gene arrays to identify gene expression profile signatures of cell/tissue subsets;

· ​RNA seq analyses (beginning in 2013) to replace gene arrays using a high-throughtput platform; and

· ​TCR clonotyping to define the repertoire of epitope (peptide) specific responses.

Our labs continue efforts to optimize and qualify a B cell ELISpot assay, the CFSE assay and the VIA.

The CTVIMC also provides CAVD investigators with a variety of peptides, avi-tagged env proteins and clinical and NHP PBMC specimens, and some NHP tissue specimens to support assay development initiatives.

​In addition to supporting the DCVAX and MUCOVAX clinical trials in Y7, the CTVIMC also provided standardized assays to 8 NHP studies, and gene array analyses and B cell Fluidigm assays to several more NHP studies.  We also maintained a robust research agenda, with specific projects focused on determining the mechanism of the VIA, interrogating TCR recognition of HIV infected cells and assessing the functional equivalent of TFH cells in the periphery.