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Using Reference Reagents to Confirm Robustness of Cytokine Release Assays for the Prediction of Monoclonal Antibody Safety
Published on: September 15, 2023
After TGN1412: recent developments in cytokine release assays.
R Stebbings1, D Eastwood, S Poole
1Biotherapeutics Group, NIBSC, Potters Bar, Hertfordshire EN6 3QG, UK. richard.stebbings@nibsc.hpa.org.uk
Journal of Immunotoxicology
|September 13, 2012
Summary
The 2006 TGN1412 trial failure highlighted regulatory science gaps. New immunotoxicity assays using human cells and specific assays like immobilized antibody or endothelial co-culture can predict adverse reactions from biological medicines.
Area of Science:
- Immunotoxicology
- Regulatory Science
- Biologics Development
Background:
- The 2006 TGN1412 monoclonal antibody (mAb) trial failure revealed limitations in standard preclinical safety testing and animal models for predicting clinical outcomes.
- Standard cynomolgus macaque models and in vitro assays failed to predict TGN1412's severe adverse effects due to differences in CD28 expression and in vivo antigen presentation.
Purpose of the Study:
- To identify the reasons for the TGN1412 immunotoxicity trial failure.
- To propose improved in vitro immunotoxicity assays that are sensitive and predictive of clinical outcomes for biological medicines.
Main Methods:
- Investigated CD28 expression differences between cynomolgus macaques and humans.
- Evaluated standard in vitro safety tests and compared them with novel assays including immobilized therapeutic mAb-based and endothelial cell co-culture assays.
- Assessed cytokine release (TNFα, IL-8, IL-2) and lymphoproliferation as potential biomarkers.
Main Results:
- CD28 is expressed on human CD4+ effector memory T-cells but not cynomolgus macaques, explaining the non-predictive animal model results.
- Standard in vitro assays did not replicate in vivo TGN1412 presentation, leading to false-negative safety predictions.
- Immobilized mAb and endothelial cell co-culture assays successfully predicted TGN1412's pro-inflammatory potential.
- IL-2 release and lymphoproliferation were identified as more discriminatory markers for TGN1412-like responses than TNFα and IL-8.
Conclusions:
- Standard preclinical and in vitro assays are insufficient for predicting the immunotoxicity of certain biological medicines like TGN1412.
- Novel in vitro assays that better mimic in vivo conditions, such as immobilized mAb or endothelial cell co-culture assays, are crucial for accurate safety assessment.
- Developing sensitive and predictive immunotoxicity assays is essential for advancing the safe development of novel biological therapeutics and preventing future trial disasters.
