Effects of intermittent T-cell cluster disaggregation on proliferative capacity and checkpoint marker expression

Abstract

Background/aim: T-cell immunotherapies are rapidly gaining grounds in clinical success. Presently, there is first-to-market knowledge on the translation of research scale methods to clinical and commercial scales. Improved understanding can lead to more consistent and efficient production, scaling, and eventual potency. T-cell checkpoint markers, proliferation, and T-cell cluster size and disaggregation are one set of parameters that have yet to be explored.

Methods: We herein activated T-cells and assessed various mechanical dissociation frequencies in relation to expression of checkpoint markers (measured by flow cytometry).

Results: We herein find increased T-cell proliferation capacity with increased dissociation frequency. We also find that with increased cluster size and duration, lower proliferation, and increased expression of checkpoint markers.

Conclusions: These findings reveal new translation findings with respect to T-cell handling and production and suggest that T-cell disaggregation may be important to improved cell yields and phenotype.

Keywords:

• T-cell
• immunotherapy
• biomanufacturing
• checkpoint
• PD1
• CTLA4
• TIM3
• Lag3
• TIGIT

Author contributions

ML and BP conceived and designed the study. ML, LC, SS performed experiments, analysed, and interpreted the data. MVM provided expert guidance on T-cell manufacturing protocols. ML and BP wrote the manuscript.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported in part by the Shriners Hospitals for Children (BP) and the National Institutes of Health Grants [R01EB012521 and T32-EB016652 (ML)].