Clarifying T Cell Mechanism in the TME in Preclinical Development

Use IsoCode profiling to elucidate how checkpoint combinations and vaccines affect T cell functionality across various tumor types.

IsoCode Data

FOCiS 2018 Abstract: Single-Cell Highly Multiplexed Proteomics Identifies Novel Polyfunctional Human CD8+ T Cell Signatures Induced by a Nanoparticle-based Melanoma Vaccine in Human Immune System Mice

Polyfunctional T cells (co-secretion of 2+ proteins per single cell) are an important functional attribute of a quality human T-cell immune response to antigen. Assessment of human T-cell polyfunctional responses to tumor vaccines in humanized mice, which are reconstituted with human immune system (HIS), provides a potentially valuable translational tool for tumor vaccine candidate screening and efficacy determination pre-clinically.

We utilized 32-plex single-cell functional proteomics platform to profile human CD8+ T-cells in HIS mice induced by a nanoparticle-based melanoma vaccine (NP). HIS mice were vaccinated with different formulations of NP or vehicle. Splenic human CD8+ T-cells were isolated, stimulated with anti-human CD3/CD28 at 37°C, 5% CO2 for 24 hours and loaded into a single-cell IsoCode Chip, pre-patterned with a 32-plex antibody ELISA array per cellular microchamber. Protein secretions were analyzed from ~1000 single human CD8+ T-cells after 16-hour-on-chip incubation.

The single-cell analysis demonstrates sensitive detection of robust upregulation of polyfunctional human CD8+ T-cell subsets in NP-vaccinated HIS mice compared to controls. The enhanced polyfunctional strength index of the human CD8+ T-cells by the vaccine was driven by antitumor-associated proteins including granzyme B, IFN-γ, MIP-1α, perforin and TNF-α. Additionally, novel single-cell visualizations revealed distinct polyfunctional cell signatures with these combinatorial human cytokine secretions that drove human CD8+ T-cell response by the NP. Polyfunctional human CD8+ T-cell responses sensitively demonstrated successful induction of the tumor vaccine in HIS mice, which may serve as a translational tool for more accurate evaluation of tumor vaccine efficacy in a pre-clinical setting.


Lu Y, Xue Q, Eisele MR, Sulistijo E, Brower K, Han L, Amir ED, Pe’er D, Miller-Jensen K, and Fan R. Highly Multiplexed Profiling of Immune Effector Functions Reveals Deep Functional Heterogeneity in Response to Pathogenic Ligands. Proc. Natl. Acad. Sci., 112 (7), 607-615 (2015).

Lu Y, Chen JJ, Mu L, Xue Q, Wu Y, Wu PH, Li J, Vortmeyer AO, Miller-Jensen K, Wirtz D, and Fan R. High-throughput Secretomic Analysis of Single Cells to Assess Functional Cellular Heterogeneity. Analytical Chemistry, 85 (4), 2548– 2556 (2013).

Ma C, Fan R, Ahmad H, Shi Q, Comin-Anduix B, Chodon T, Koya RC, Liu CC, Kwong GA, Radu CG, Ribas A, and Heath JR. A Clinical Microchip for Evaluation of Single Immune Cells Reveals High Functional Heterogeneity in Phenotypically Similar T Cells. Nature Medicine, 17, 738-743 (2011).

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