Predict Immunotoxicological Response

Therapeutics that engage and modulate immune targets represent a significant and growing component of candidate drugs in discovery and development. A challenge presented by this cohort of drugs is that while modulation of immune function can provide profound therapeutic benefit, they may also trigger adverse events due to undesired impacts on immune function. There is a requirement that during development, the potential immunotoxicology of these therapeutics is thoroughly investigated and their risk profile established. Charles River specializes in mapping human immune function in vitro, ex vivo, and in vivo from early development and lead selection through to clinical trial support. We can work with you to test predictive immunotoxicology and reduce the risk that this will impact your therapeutic.

Antibodies to cell movement of the virus

Have You Fully De-Risked Your Candidate?

Read about several approaches to predictive immunotoxicology, highlighting different types of assays and how they are used to evaluate the risks of enhanced on-target effects and reveal the unexpected.

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Regulatory Expectations

Regulatory authorities expect that the potential for immunotoxicity of all immunomodulatory therapeutics is appropriately tested. These predictive immunotoxicology assessments, based on the risk profile derived from the mode of action, may extend through in vitro assessments such as the cytokine release assays (CRA assays), specific endpoints incorporated into pre-clinical studies, and monitoring of immune function within clinical trials. The major challenge in charting a successful course through this process is working with a partner who can deliver expert scientific and regulatory support.

The range of predictive immunotoxicology of therapies are often closely tied to the desired pharmacology of the therapeutic. As such, the program of immunotoxicology assessments must be tailored and refined to individual clinical candidates.

Diagram illustrating the possible adverse drug interactions with immune cells.

diagram illustrating the possible adverse drug interactions with immune cells

We can select from a broad range primary human immune cell assays to model the diverse immunological mechanisms to help de-risk the potential immunotoxicology of novel therapeutics.

Validated Immunotoxicology Assays

We have validated primary human immunological assays that enable you to predict potential immunotoxicology during preclinical development, and in addition can support your clinical trials to determine the mechanisms underlying any observed infusion reactions.

Our extensive predictive immunotoxicology range of assays allows you to work with our scientists to build a tailored program for your therapeutic suitable for each stage of development.

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  • Infusion-related reaction assays

    These predictive immunotoxicology assays are designed to capture the risk of immunotox during, or immediately following, the infusion period. These effects are often directly related to the mode of action of a therapeutic but may have multiple underlying causes.

    • Cytokine release (CRA assays)
    • Complement activation, C3a, C5a, SC5b-9, CH50
    • IgE
    • Tryptase
    • Platelets


    t cell, immune cell, immune system, cell assays

    CRA Assays for Predictive Immunotoxicology

    The assessment of cytokine release assays (CRA assays) plays a key role in developing immunomodulatory therapeutics. Our experts for this panel discussion share their knowledge on this topic.

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  • Modification of innate immune function and differentiation

    These predictive immunotoxicology assays are designed to assess the risk of immunosuppression by non-immunomodulating drugs, as well as the mechanism of action or anticipated pharmacology of immune modulators.

    Because innate immune effectors are the body’s first line of defense against pathogens, any changes in function to NK cells or neutrophils are of keen interest. Additionally, dendritic cells and macrophages play a role in priming adaptive immune responses; therefore, any suppression of these cell populations can have detrimental downstream effects on T and B cell responses. Mast cells are also a key contributor to anaphylactic (Type I hypersensitivity) responses and may be assessed for their potential to degranulate in response to various test articles.

    • Immunophenotyping
      • Activation and cytotoxicity markers
      • Receptor occupancy
      • Intracellular cytokine staining
    • Neutrophil/macrophage function and activation
      • Oxidative burst activity
      • Phagocytosis
      • Migration
      • M1/M2 polarization
    • Cytokines and chemokines
      • Plasma/serum profiling
      • Intracellular staining
    • Cytotoxicity assays for NK cell function
    • ADCC/CDC
    • Histamine and tryptase degranulation assays (mast cells and basophils)
    • Fc-gamma receptor binding
  • Modification of adaptive immune function and differentiation

    Adaptive immune effectors provide potent activity against invading pathogens and provide immune memory. A variety of test articles have been developed to activate (cancer immunotherapies) or suppress (autoimmunity) the function of T cell and B cell populations.

    The predictive immunotoxicology assays below provide the means for characterizing the effect of a test article on these cell populations and cover a broad range of endpoints. Cell surface and intracellular markers can be used to characterize the phenotypes of T cell and B cell populations. Functional assays capture the modulation of key activities of these cell types: cytotoxicity, proliferation, CRA assays, antibody responses, and the assessment of antigen specific responses.

    • Immunophenotyping
      • Broad range of validated panels are available
      • Custom receptor occupancy assays can be developed and validated
      • TNBK panels
      • Treg and subset analysis
      • Proliferation and intracellular cytokine profiling
      • Activation and cytotoxicity markers


    Flow cytometry used for immunophenotyping

    Validating Flow Cytometry Assay Panels for Blood Cell Subpopulation

    This poster examines the development of different methods for predictive immunotoxicology that were successfully validated to analyze samples in a toxicology study.

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    • T-cell cytotoxicity
    • ELISPOT analysis for antigen specific T cell responses
    • T cell dependent antibody responses (TDAR)
    • Delayed type hypersensitivity (DTH)
    • Cytokines and chemokines
      • Plasma/serum profiling
      • PBMC/ whole blood CRA assay
    • T cell Polarization – Th1, Th2, Th17
      • Analysis of target gene signature

Our scientists can use whole blood and PBMC, but also have extensive experience working with purified populations (e.g., T cells, dendritic cells, macrophages, platelets, and endothelial cells) to conduct predictive immunotoxicology assays.

bar graph showing PBMC cytokine release in response to therapeutic antibodies

Example: Novel biologics and immune modulating drugs can be benchmarked against a range of therapeutic antibodies with known toxicology profiles for their ability to drive CRA assays in whole blood and PBMC assays.

Our experience with the broad range of predictive immunotoxicology assays provides you the tools needed to develop your therapeutic. From monitoring and assessing the desired pharmacology, to performing early assessments of when that desired pharmacology may cross over into immunotoxicology risk, by selecting the right assays for the target and therapeutic type, we can reduce risks for your programs and your patients.

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Frequently Asked Questions (FAQs) about Predictive Immunotoxicology

  • For which therapeutics can predictive immunotoxicology assays be applied?

    While immunotoxicology assessments are broadly applied to therapeutics with a targeted immunomodulatory function, they can also be applied to those where there is a class risk.

  • When should I apply this strategy to my development?

    As a component of a ‘fail early, fail cheaply’ approach, it is important to apply appropriate risk mitigation strategies before lead selection. An effective approach to fully de-risking drug candidates with predictive assays in the earlier stages of development can help to avoid failure later in the clinic.

  • How should an appropriate group of analysis be selected?

    As there are a wide range of assays that can speak to the risk of immunotoxicity, the best starting point is to consider the pharmacology of the drug. This is especially relevant to monoclonal antibodies, and in this case both the target and the activities of the Fc region should be considered.

  • For qualitative assays such as CRA, how should data be interpreted?

    The degree of cytokine release is often directly related to the pharmacology of the test article. Determining to what degree the results represent an identification of a clinical risk will depend on the inclusion of appropriate controls, selection of a suitable number of donors, and detailed scientific review of the results.