Table 2 Immunodynamic endpoint assessment
Immuno-prognostic Immunotherapeutic Area | Immunodynamic Endpoint | Method of Assessment | Site of Assessment |
|---|---|---|---|
Prognostic | |||
Immunoscore | T-cell infiltrate | IHC: 1. CD8 2. CD45RO | Tumor |
Therapeutic | |||
Conventional Therapies | Â | Â | Â |
Chemotherapy | Immunogenic Cell Death | IHC: 1. phosphorylated eIF2α 2. nuclear HMGB1 (late apoptosis-related marker) 3. LC3-B (autophagosome-related marker) 4. Mx1 and TLR3 (IFN signature) 5. CD8/Foxp3 or CD8/CD68 ratios | Tumor |
| Â | Â | Gene expression analyses: 1. Cxcl10 2. IFNb 3. TLR3 | Tumor |
Radiation therapy | Tumor immunogenicity | Gene expression analysis: 1. Immunologic constant region (ICR) | Tumor |
| Â | Radiation induced T and NK activation | IHC: 1. MICA | Tumor |
| Â | Â | ELISPOT: 1. sMICA 2. anti-sMICA antibodies | Peripheral Blood - Serum |
| Â | Memory T cell response | T cell receptor (TCR) repertoire analysis | PBMCs |
ITAs - Passive | Â | Â | Â |
Cellular Therapy | Â | Â | Â |
Adoptive T and NK cells | Quantification of adoptive cell population | Flow cytometry–based or PCR-based assessment of unique label in adoptive cell population | PBMCs |
Chimeric Antigen Receptor (CAR) T cells | Phenotype of adoptive cell population | Flow cytometry based or PCR based assessment of phenotype of adoptive cell population (see Tables 3 and 4; activation and inhibitory markers including ICOS, 4-1BB, PD-1, PD-L1, OX-40, LAG-3, GITR, VISTA, LIGHT) | PBMCs |
|  | Function of adoptive cell population | Flow cytometry based (perforin, granzyme, intracellular cytokine expression including IFN-γ), ELISPOT, in-vitro cytotoxicity (ie chromium release) or PCR based assessment of phenotype of adoptive cell population after antigen-specific (preferred, ie autologous tumor cells or tumor peptide pulsed T2 cells) or nonspecific stimulation (ie CD3/CD28, PMA, or ionomycin) | PBMCs |
| Â | Toxicity | Comprehensive cytokine assessment, CRP | Peripheral Blood - Serum or plasma |
ITAs - Active & Specific | Â | Â | Â |
Monoclonal Antibodies | Â | Â | Â |
Tumor-targeting | Target antigen expression in tumor | IHC, multicolor IF, and/or in-situ gene expression of target antigen of mAb on tumor (CD20, HER2, EGFR) | Tumor (primary, metastatic and circulating disease) |
|  | FcR polymorphism | FcR genotype (FcgRIIIA, FcgRIIA) | Peripheral blood – Mononuclear cells (genomic DNA) |
|  | Phenotype and function of immune (T and NK cell) response | T cell receptor (TCR) repertoire analysis Flow cytometry based or PCR based assessment of phenotype of T and NK cells (see Tables 3 and 4) and flow cytometry based (perforin, granzyme, intracellular cytokine expression including IFN-γ), ELISPOT, in-vitro cytotoxicity (ie chromium release) or PCR based assessment of phenotype T and NK cells after antigen-specific (preferred, ie autologous tumor cells or tumor peptide pulsed T2 cells) or nonspecific stimulation (ie CD3/CD28, PMA, or ionomycin) | PBMCs |
|  | Systemic cytokine response | Comprehensive cytokine assessment (IL-6, IFN-γ, IL-10) and serologic assessment (sIL-2Rα, MCP) | Peripheral Blood - Serum or plasma |
Immune-targeting, including checkpoint inhibitors | Target expression in tumor microenvironment | IHC, multicolor IF, and/or in-situ gene expression of target on tumor and stroma (CTLA-4, PD-1, PD-L1, OX-40, 4-1BB, LAG-3, GITR, CD40) | Tumor |
| Â | Tumor infiltrating immune response | IHC: 1. Quantity and phenotype of tumor-infiltrating lymphocytes (TILs) 2. CD8 effector: CD4 regulatory T cell ratio | Tumor |
| Â | Memory T cell response | T cell receptor (TCR) repertoire analysis | PBMCs |
| Â | Clinical response | CRP, LDH, WBC, ALC, MDSCs | Peripheral Blood - Serum or plasma |
| Â | Toxicity | Comprehensive cytokine assessment | Peripheral Blood - Serum or plasma |
Vaccines | Â | Â | Â |
In-situ Vaccines Cell-based Vaccines | Target antigen expression in tumor | IHC, multicolor IF, and/or in-situ gene expression of target vaccination antigen on tumor (gp100, MART, Mucin) | Tumor |
 Dendritic Cell-based  Vaccines Non–cell-based Vaccines | T cell response postvaccination in tumor | IHC, multicolor IF (pre/post assessment of ratio of Treg to Teffectors and CD1a, CD8, CD94; CD207 and HLA-DR), and/or in-situ gene expression of intratumoral T cell population | Tumor |
| Â | Quantification of T cell response | T cell receptor (TCR) repertoire analysis Flow cytometry based or PCR based assessment of tumor-specific T cells (dimer, tetramer, dextramer) | PBMCs) |
|  | Phenotype of T cell response | T cell receptor (TCR) repertoire analysis Flow cytometry based or PCR based assessment of phenotype of tumor-specific T cells (see Table 3) | PBMCs) |
|  | Function of T cell response | Flow cytometry based (perforin, granzyme, intracellular cytokine expression including IFN-γ), ELISPOT, in-vitro cytotoxicity (ie chromium release) or PCR based assessment of phenotype of tumor-specific T cells after antigen-specific (preferred, ie autologous tumor cells or tumor peptide pulsed T2 cells) or nonspecific stimulation (ie CD3/CD28, PMA, or ionomycin) | PBMCs |
| Â | Humoral response Systemic cytokine response | ELISPOT tumor/antigen antibody response Comprehensive cytokine assessment (GM-CSF) | Peripheral Blood - Serum or plasma |
ITargeted Agents Active & Nonspecific | Â | Â | Â |
Cytokines | Intratumoral immune response | IHC, multicolor IF, and/or in-situ gene expression of intratumoral lymphocyte (T, B, and NK cell) population | Tumor |
|  | Phenotype and function of immune(T and NK) response | T cell receptor (TCR) repertoire analysis Flow cytometry based or PCR based assessment of phenotype of T and NK cells (see Tables 3 and 4) and flow cytometry based (perforin, granzyme, intracellular cytokine expression including IFN-γ), ELISPOT, in-vitro cytotoxicity (ie chromium release) or PCR based assessment of phenotype T and NK cells after antigen-specific (preferred, ie autologous tumor cells or tumor peptide pulsed T2 cells) or nonspecific stimulation (ie CD3/CD28, PMA, or ionomycin) | PBMCs |
|  | Systemic serologic and cytokine response | Comprehensive cytokine assessment (IL-6, IFNγ, IL-10) and serologic assessment (sIL-2Rα, MCP) | Peripheral Blood - Serum or plasma |
IDO Inhibitors | IDO expression in tumor | IHC and/or in-situ gene expression of IDO1 | Tumor |
| Â | Inhibition of IDO1 based on Kyn/Trp ratio | Kyn/Trp level and IHC of DC maturation status (CD80, CD86) | Tumor & Peripheral Blood - Serum or plasma |
|  | Phenotype and function of immune (T and NK cell) response | T cell receptor (TCR) repertoire analysis Flow cytometry based or PCR based assessment of phenotype of T and NK cells (see Tables 3 and 4) and flow cytometry based (perforin, granzyme, intracellular cytokine expression including IFN-γ), ELISPOT, in-vitro cytotoxicity (ie chromium release) or PCR based assessment of phenotype T and NK cells after antigen-specific (preferred, ie autologous tumor cells or tumor peptide pulsed T2 cells) or nonspecific stimulation (ie CD3/CD28, PMA, or ionomycin) | PBMCs |