Immunotoxicology

PBMC cytokine release assay (CRA) – T cell immunogenicity risk panel

Background information

  • The development of anti-drug antibodies (ADA) towards peptide drugs can alter their bioavailability and pharmacokinetics in vivo resulting in a reduction in overall drug efficacy.1
  • The presence of ADA also negatively impacts the safety of these therapeutics by increasing the likelihood of adverse reactions being observed in patients to whom the drug is administered.2
  • While ADA generation can occur via B-cell dependent pathways,3 the immunogenic response of biotherapeutics is predominantly T-cell mediated and governed by epitopes found in their primary sequences.4
  • Plate-based approaches centring on evaluating the immunogenic risk posed by therapeutic monoclonal antibodies (mAbs) have identified IL-2 as the most predictive cytokine readout,5 although IFN-γ was also shown to have appreciable utility in this regard.5-6
  • Our standard T-cell immunogenicity risk panel measures the release of IL-2 & IFN-γ released by PBMC upon exposure to test articles 6 days post challenge.
  • We have demonstrated the utility of this panel by screening a selection of mAbs whose immunogenicity risks have been widely documented in the literature (Table 1).
  • Our standard assay uses Phytohemagglutinin-L (PHA-L) as a positive control to induce the release of cytokines associated with T-cell activation. When screened as part of our CRA, cellular exposure to PHA-L resulted in an increase in the levels of both IL-2 and IFN-γ (Fig 1).

Protocol

PBMC Proliferation Protocol

Data

Data from Cyprotex – T-cell immunogenicity risk panel.

References

1) De Groot AS & Scott DW, (2007). Immunogenicity of protein therapeutics. Trends Immunol., 28; 482–90.
2) Hansel TT et al., (2010). The safety and side effects of monoclonal antibodies. Nat. Rev. Drug Discov., 9; 325-38.
3) Vaisman-Mentesh et al., (2020). The Molecular Mechanisms That Underlie the Immune Biology of Anti-drug Antibody Formation Following Treatment With Monoclonal Antibodies. Front. Immunol., 11, 1951.
4) Jawa V et al., (2020). T-Cell Dependent Immunogenicity of Protein Therapeutics Pre-clinical Assessment and Mitigation–Updated Consensus and Review. Front. Immunol., 11:1301.
5) Joubert MK et al., (2016). Use of In Vitro Assays to Assess Immunogenicity Risk of Antibody-Based Biotherapeutics. PLoS One. 11 (8); e0159328.
6) Rombach-Riegraf V et al., (2014). Aggregation of human recombinant monoclonal antibodies influences the capacity of dendritic cells to stimulate adaptive T-cell responses in vitro. PLoS ONE. 9; e86322.

TODO

Cyprotex eStore

Order our services online.

Visit the Cyprotex eStore
Sam Bevan

Sam Bevan

Principal Scientist

vCard
Logo Cyprotex white
Cyprotex enables and enhances the prediction of human exposure, clinical efficacy and toxicological outcome of a drug or chemical. By combining quality data from robust in vitro methods with contemporary in silico technology, we add value, context and relevance to the ADME-Tox data supplied to our partners in the pharmaceutical or chemical industries.