Accelerating precision medicine™

Immune Monitoring in Oncology

Background in Oncology

Cancer is one of the leading causes of death in industrialized countries. Immuno-oncology is an innovative new field of cancer research and focuses on treatment that aims to harness the body’s immune system to control tumor growth.

Immunotherapy strategies include immunization to cancer antigens (cancer vaccines), alteration of local immunosuppressive signals to favor an anti-tumor response (checkpoint blockade), and manipulation of patient cells ex vivo to amplify their ability to kill tumor cells when re-injected into patients (adoptive cell therapy).

Immune Monitoring Expertise in Oncology

ImmuneCarta has the expertise and tools to phenotypically and functionally characterize the immune response to tumor cells in tissue through the isolation of Tumor-Infiltrating Lymphocytes (TIL) or in peripheral blood. Our diverse multiparametric immune monitoring assays can be applied to a wide range of studies and therapeutic approaches.

  • Adoptive Cellular Therapies (Dendritic cell vaccination, Chimeric Antigen Receptor (CAR) T cells)

  • Studies analyzing the clinical response following immune checkpoint blockade (αPD-1, αCTLA-4, αPD-L1)
    • Development of surrogate compound screening assays for prioritizing immune modulating drugs
  • Studies which aim to analyze the local tumor microenvironment
    • TIL isolation and characterization of the immunosuppressive compartment
    • Measurement of TIL functionality in the presence and absence of immune modulators
  • Cancer vaccine immunotherapy and measurement of tumor-specific T cell frequency and functionality

Checkpoint Blockade (Inhibitor) Immunotherapy

Background in Checkpoint Blockade (Inhibitor) Immunotherapy

An efficient immune response requires that a fine balance be established between co-stimulatory and co-inhibitory signals. This balance regulates effector responses and assures immune protection while limiting uncontrolled inflammation.

It is now well-established that tumors and viruses co-opt immune checkpoint receptors (e.g. PD-1, CTLA-4, Tim-3, LAG-3) as a mechanism of immune evasion. Checkpoint inhibitors have yielded positive clinical outcomes by rescuing cancer and virus-specific exhausted T cells resulting in an enhanced immune response.

Expertise in Checkpoint Blockade Immunotherapy

ImmuneCarta has extensive expertise in the field of checkpoint blockade and offers surrogate assays for compound screening of checkpoint inhibitors.

We develop highly multiplexed flow cytometry panels to analyze immune exhaustion. Carefully designed panels provide information on the relative expression of co-inhibitory molecules, as well as the proportion of recently activated to fully exhausted T cells.

ImmuneCarta’s functional assays (ICS and CFSE) assess the ability of novel compounds to reverse functional exhaustion of T cells and provide clients with a comprehensive view of the immune status.

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Surrogate Assays for Screening Immune Modulating Drugs

Background of Surrogate Assays in Determining Drug Efficacy

ImmuneCarta uses T cells isolated from HIV-infected patients to assess the efficacy of immune checkpoint blockade in vitro by measuring the functional impact on HIV-specific T cells.

  • HIV-specific T cells co-express immune checkpoints
  • Peripheral T cells are easily accessible for in vitro compound testing

Expertise in Surrogate Assays

Leukapheresis samples from chronic HIV-infected patients are obtained and PBMC (peripheral blood mononuclear cells) are isolated and cryopreserved. After identification of breadth, magnitude and specificity of response using a matrix ELISpot (Enzyme-Linked Immunosorbent Spot ) approach, PBMC are stimulated with HIV peptides in the presence and absence of immune checkpoint molecules.

Proliferation utilizing CFSE (Carboxyfluorescein succinimidyl ester) and cytokine secretion utilizing ICS (Intracellular Cytokine Staining) are measured in the presence and absence of immune modulators.

  • Efficacy of inhibitors is determined by an increase in proliferation and cytokine secretion.

Flow Cytometry dot plots depicting immune response of T cells in presence of immunotherapy

Dot plots showing the frequency of proliferating HIV-specific CD3 T cells in the presence of an immune modulating compound

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Adoptive Cell Therapy of Chimeric Antigen Receptor (CAR) T cells

Background in Adoptive Cell Therapy

CAR T cells are detected by flow cytometry in PBMC (peripheral blood mononuclear cells) isolated from treated patients using HLA-restricted multimers. Functionality is also assessed using intracellular cytokine staining (ICS) and/or CFSE proliferation.

Immune response monitoring for adoptive cell therapy when patients infused with own stimulated T cells

Adoptive cell therapy involves
stimulating patient’s T cells in vitro,
and infusing patient with stimulated cells

The functional and phenotypic profile of these cells at pre-infusion and post-infusion time points provides immune response information on the fate and persistence of these adoptively transferred effector T cells.

Expertise in Adoptive Cell Therapy

ImmuneCarta uses phenotypic and functional profiling to confirm antigen specificity and functionality of cells for adoptive cell therapy.

We monitor peripheral immune responses to therapeutic vaccines formulated with autologous dendritic cells pulsed with viral or tumor antigens, and have developed standardized procedures for the generation and maturation of monocyte-derived dendritic cells (MDDC) for in vitro stimulation assays.

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Cancer Vaccine Immunotherapy


Cancer vaccine immunotherapy has garnered much interest and success in recent years. These therapeutic vaccines are designed to train the immune system to recognize and attack cancer cells

Expertise in Cancer Vaccine Immunotherapy

  • ImmuneCarta has the expertise and tools to phenotypically and functionally characterize and monitor the immune response to cancer antigens following immunization.
    • ImmuneCarta successfully collaborated with ImmunoVaccine in the DPX-Survivac clinical trial. Results were presented in a poster presentation at the 2014 AACR Conference in April, entitled "Immune profiling of regulatory T cells, frequency of Survivin-specific T cells and Intracellular cytokine Staining (ICS) in ovarian cancer patients vaccinated with a therapeutic vaccine."
    • Key finding: Phenotypic and functional probing demonstrated durable and polyfunctional CD4+ and CD8+ T cell responses

"The flow cytometry-based immune monitoring performed by ImmuneCarta has added significant value to our clinical program by clearly demonstrating the mechanism of action of our DPX-Survivac vaccine in cancer patients."
Marc Mansour, Ph.D., Chief Operating Officer of ImmunoVaccine

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