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Table 2 Immunodynamic endpoint assessment

From: Immunodynamics: a cancer immunotherapy trials network review of immune monitoring in immuno-oncology clinical trials

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    
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
   Gene expression analyses: 1. Cxcl10 2. IFNb 3. TLR3 Tumor
Radiation therapy Tumor immunogenicity Gene expression analysis: 1. Immunologic constant region (ICR) Tumor
  Radiation induced T and NK activation IHC: 1. MICA Tumor
   ELISPOT: 1. sMICA 2. anti-sMICA antibodies Peripheral Blood - Serum
  Memory T cell response T cell receptor (TCR) repertoire analysis PBMCs
ITAs - Passive    
Cellular Therapy    
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
  Toxicity Comprehensive cytokine assessment, CRP Peripheral Blood - Serum or plasma
ITAs - Active & Specific    
Monoclonal Antibodies    
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
  Tumor infiltrating immune response IHC: 1. Quantity and phenotype of tumor-infiltrating lymphocytes (TILs) 2. CD8 effector: CD4 regulatory T cell ratio Tumor
  Memory T cell response T cell receptor (TCR) repertoire analysis PBMCs
  Clinical response CRP, LDH, WBC, ALC, MDSCs Peripheral Blood - Serum or plasma
  Toxicity Comprehensive cytokine assessment Peripheral Blood - Serum or plasma
Vaccines    
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
  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
  Humoral response Systemic cytokine response ELISPOT tumor/antigen antibody response Comprehensive cytokine assessment (GM-CSF) Peripheral Blood - Serum or plasma
ITargeted Agents
Active & Nonspecific
   
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
  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
  1. ALC absolute lymphocyte count, CRP C-reactive protein, DC dendritic cell, IDO indoleamine-2,3-dioxygenase; IFN-γ interferon gamma, LDH lactate dehydrogenase, MCP Monocyte Chemotactic Protein-1; MDSC myeloid-derived suppressor cells, PBMC peripheral blood mononuclear cells, sIL-2Rα, soluble IL-2 receptor-alpha, WBC white blood cell count