Biological medicines can potentially cause adverse drug reactions related to immunosuppression, immunostimulation and hypersensitivity.
These include cytokine release-associated acute reactions involving the production of pro-inflammatory cytokines. The clinical manifestations can range from flu-like reactions to more severe cytokine release syndromes (CRS).
In 2006 a severe CRS was observed during the clinical trial of the therapeutic anti-CD28 monoclonal antibody (mAb) TGN1412 – developed to treat B cell leukaemia and autoimmune diseases.
We were involved in testing trial materials and understanding why pre-clinical safety testing failed to predict the adverse effects observed in humans.
Our findings have been used to develop improved pre-clinical testing methods for biological medicines. Find out more in research published in the Journal of Immunology.
Although severe incidences like this are rare, correctly predicting the cytokine-releasing potential of new biotherapeutics is a major safety concern.
Disseminated intravascular coagulation experienced by a volunteer during the 2006 TGN1412 clinical trial.
Use of intracellular cytokine staining to study the different mechanisms of cytokine release by therapeutic mAbs – Identification of immune cell subpopulation involvement in IFNγ production
We provide contract pre-clinical safety testing of biological medicines and vaccines. We use state-of-the-art in vitro cytokine release assays considered to reflect the ‘cytokine storm’ that occurred during the phase I clinical trial of TGN1412.Find out more about NIBSC control testing. Contact us: Sandrine.Vessillier@nibsc.org
To harmonize the field of immunotoxicology evaluation, we have developed a panel of validated reference reagents for cross-platform comparison of cytokine release data. This panel (NIBSC code: 19/156) consists of three isotype controls plus three positive controls (human anti-CD52, mouse anti-CD3 and human superagonistic anti-CD28 mAb ) with different mechanisms of action and inducing cytokine release from different type of immune cells.
Our comparative study of cytokine release assays highlighted the strengths and weaknesses of different protocols used in pre-clinical studies. We found that a “one size fits all” approach for the pre-clinical safety testing of biotherapeutics isn’t suitable and recommend a tiered approach to detect multiple mechanisms of cytokine release.
We showed that the presence of red blood cells hampers the induction of T cell-derived cytokines and recommend using isolated white blood cells for evaluation of cytokine release by T cells.
However whole blood cell assays are more predictive for evaluation of non-T cell-derived cytokines.
Glycophorin A (GYPA), the main protein on erythrocytes inhibited TGN1412-induced cytokine release: Cytokine release after red blood cell (RBC) depletion (RBC depleted) in presence of glycophorin A (+ GYPA) or whole blood (+ WB)
Vessillier, S., M. Fort, L. O’Donnell, H. Hinton, K. Nadwodny, J. Piccotti, P. Rigsby, K. Staflin, R. Stebbings and D. Mekala, Willingham, A. and B. Wolf (2020). "Development of the first reference antibody panel for qualification and validation of cytokine release assay platforms-Report of an international collaborative study." Cytokine: X: 100042. https://doi.org/10.1016/j.cytox.2020.100042
Whitfield, S. J. C. , Taylor, C., Risdall, J. E., Griffiths, G. D. , Jones, J. T. A., Williamson, E. D., Rijpkema, S., Saraiva, L. , Vessillier, S., Green , A. C. and Carter, A. J. (2017) Interference of the T Cell and Antigen-Presenting Cell Costimulatory Pathway Using CTLA4-Ig (Abatacept) Prevents Staphylococcal Enterotoxin B Pathology. The Journal of Immunology, 198 (10): 3989-3998. https://doi.org/10.4049/jimmunol.1601525Vessillier, S., Eastwood, D., Fox, B., Swaminathan, S., Sathish, J. Thorpe, S.J., Thorpe, R. and Stebbings, R. (2015) Cytokine release assays for the prediction of therapeutic mAb safety in first-in man trials - Whole blood cytokine release assays are poorly predictive for TGN1412 cytokine storm. Journal of immunological methods 424:43-52. https://doi.org/10.1016/j.jim.2015.04.020Reed, D.M., Paschalaki, K.E., Starke, R.D., Mohamed, N.A, Sharp, G., Fox, B., Eastwood, E., Bristow, A., Ball, C., Vessillier, S., Hansel, T.T., Thorpe, S.J., Randi, A.M., Stebbings, R. and Mitchell, J.A. (2015) An autologous endothelial cell: PBMC assay that detects cytokine storm responses to biologics. FASEB Journal, 29(6): 2595-602. https://doi.org/10.1096/fj.14-268144Stebbings, R., L. Findlay, C. Edwards, D. Eastwood, C. Bird, D. North, Y. Mistry, P. Dilger, E. Liefooghe, I. Cludts, B. Fox, G. Tarrant, J. Robinson, T. Meager, C. Dolman, S. J. Thorpe, A. Bristow, M. Wadhwa, R. Thorpe and S. Poole (2007). "Cytokine storm" in the phase I trial of monoclonal antibody TGN1412: better understanding the causes to improve preclinical testing of immunotherapeutics. J.Immunol. 179(5): 3325-3331. https://doi.org/10.4049/jimmunol.179.5.3325Eastwood D, Findlay L, Poole S, Bird C, Wadhwa M, Moore M, Burns C, Thorpe R, Stebbings R. (2010). Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4+effector memory T-cells. Brit. J. Pharmacol. 161:512-26. https://doi.org/10.1111/j.1476-5381.2010.00922.xEastwood D, Bird C, Dilger P, Findlay L, Poole S, Thorpe SJ, Thorpe R, Wadhwa M, Stebbings R. (2013). Severity of the TGN1412 trial disaster cytokine storm correlated with IL-2 release. Brit. J. Clin. Pharmacol. 76:299-315. https://doi.org/10.1111/bcp.12165Stebbings R, Eastwood D, Poole S, Thorpe R. (2013). After TGN1412: Recent developments in cytokine release assays. J. Immunotoxicology 10:75-82. https://doi.org/10.3109/1547691X.2012.711783
Dr Sandra Diebold, Section leader Dr Sandrine Vessillier, Head of ImmunotoxicologyDeepa Rajagopal Elliot Macleod