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In Vitro Assay to Evaluate the Impact of Immunoregulatory Pathways on HIV-specific CD4 T Cell Effector Function
Published on: October 15, 2013
Preclinical immunogenicity risk assessment of biotherapeutics using CD4 T cell assays
Robin E Walsh1, Angela Nix1, Chloé Ackaert2
1Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, United States.
Abstract:
T-cell dependent antibody responses to biotherapeutics remain a challenge to the optimal clinical application of biotherapeutics because of their capacity to impair drug efficacy and their potential to cause safety issues. To minimize this clinical immunogenicity risk, preclinical assays measuring the capacity of biotherapeutics to elicit CD4 T cell response in vitro are commonly used. However, there is considerable variability in assay formats and a general poor understanding of their respective predictive value. In this study, we evaluated the performance of three different CD4 T cell proliferation assays in their capacity to predict clinical immunogenicity: a CD8 T cell depleted peripheral blood mononuclear cells (PBMC) assay and two co-culture-based assays between dendritic cells (DCs) and autologous CD4 T cells with or without restimulation with monocytes. A panel of 10 antibodies with a wide range of clinical immunogenicity was selected. The CD8 T cell depleted PBMC assay predicted the clinical immunogenicity in four of the eight highly immunogenic antibodies included in the panel. Similarly, five antibodies with high clinical immunogenicity triggered a response in the DC: CD4 T cell assay but the responses were of lower magnitude than the ones observed in the PBMC assay. Remarkably, three antibodies with high clinical immunogenicity did not trigger any response in either platform. The addition of a monocyte restimulation step to the DC: CD4 T cell assay did not further improve its predictive value. Overall, these results indicate that there are no CD4 T cell assay formats that can predict the clinical immunogenicity of all biotherapeutics and reinforce the need to combine results from various preclinical assays assessing antigen uptake and presentation to fully mitigate the immunogenicity risk of biotherapeutics.
Insights
Preclinical CD4 T cell assays show limited ability to predict biotherapeutic immunogenicity. Combining multiple assays is crucial for mitigating risks associated with biotherapeutics.
Area of Science:
- Immunology
- Biotherapeutics Development
- Preclinical Assays
Background:
- T-cell dependent antibody responses to biotherapeutics pose clinical challenges, impacting efficacy and safety.
- Preclinical CD4 T cell assays are used to predict immunogenicity, but assay variability and poor predictive value are concerns.
Purpose of the Study:
- To evaluate the predictive performance of three distinct CD4 T cell proliferation assays for biotherapeutic clinical immunogenicity.
- To assess the utility of different assay formats in identifying potential immunogenic biotherapeutics.
Main Methods:
- Evaluated a CD8 T cell depleted peripheral blood mononuclear cell (PBMC) assay.
- Assessed two co-culture assays: dendritic cells (DCs) with autologous CD4 T cells, with and without monocyte restimulation.
- Used a panel of 10 antibodies with varying clinical immunogenicity.
Main Results:
- The CD8 T cell depleted PBMC assay predicted immunogenicity for 4 out of 8 highly immunogenic antibodies.
- DC:CD4 T cell assays detected responses in 5 high-immunogenicity antibodies, but with lower magnitude than PBMC assays.
- Three highly immunogenic antibodies elicited no response in either assay format.
Conclusions:
- No single CD4 T cell assay format reliably predicts clinical immunogenicity for all biotherapeutics.
- Combining diverse preclinical assays, including those for antigen uptake and presentation, is essential for comprehensive immunogenicity risk mitigation.

