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Published on: November 4, 2016
Optimizing canine T cell activation, expansion, and transduction
Treyvon W Davis1, Jennifer C Holmes2, Arissa He3
1Department of Biological Sciences, College of Veterinary Medicine, North Carolina State University, North Carolina, Raleigh, United States of America.
Abstract:
Dogs are becoming an important model for human cancers, and successfully troubleshooting issues with genetically modified T cell immunotherapy for round cell and solid neoplasms in dogs provides a unique opportunity to improve efficacy, safety, and affordability for humans as well. Unfortunately, T cell activation in dogs for optimal viral transduction has not been determined, restricting advancements in canine T cell immunotherapy. Two αCD3 and two αCD28 antibody clones for canine T cell stimulation have been described in the literature, but no studies have been undertaken to evaluate which αCD3/αCD28 combination is most effective, nor has anyone directly compared the efficacy of the two most popular antibody presentation strategies: antibody-coated plates and antibody-conjugated beads. In evaluating the effects of plate- or bead-bound αCD3 stimulation alone versus αCD3/αCD28 in combination, we tested 12 possible antibody stimulation strategies in addition to evaluating two largely unexplored mitogens in canine T cell transduction, phorbol myristate acetate (PMA) with ionomycin and concanavalin A (ConA). We investigated the impact of these stimulation strategies on canine T cell activation, expansion, and transduction. For stimulation strategies producing the best results, we also examined how each strategy affected the proportions of CD4/CD8 T cell subsets and regulatory T cell (Treg) prevalence. We determined that, in general, plate-bound antibodies were far superior to bead-bound antibodies for canine T cell stimulation, and that plate-bound αCD3 clone CA17.6F9 in combination with αCD28 clone 5B8 or the mitogen PMA with ionomycin produced better activation and expansion profiles, better transduction, and more desirable T cell subsets that are more likely to improve patient outcomes in dogs suffering from round cell and solid tumors.
Insights
Optimizing canine T cell immunotherapy requires effective T cell activation. Plate-bound antibodies, specifically αCD3 clone CA17.6F9 with αCD28 clone 5B8 or PMA/ionomycin, enhance T cell activation, expansion, and transduction for better cancer treatment outcomes in dogs.
Area of Science:
- Immunology
- Veterinary Oncology
- Cell Therapy
Background:
- Dogs serve as valuable models for human cancer research.
- Canine T cell immunotherapy shows promise but is limited by suboptimal T cell activation and transduction methods.
- Existing research lacks a comprehensive comparison of different antibody stimulation strategies and presentation methods for canine T cells.
Purpose of the Study:
- To determine the most effective methods for canine T cell activation, expansion, and viral transduction for immunotherapy.
- To compare the efficacy of plate-bound versus bead-bound antibodies for T cell stimulation.
- To evaluate different antibody clone combinations and mitogens for optimizing canine T cell responses.
Main Methods:
- Tested 12 antibody stimulation strategies, including combinations of two αCD3 and two αCD28 antibody clones presented on plates or beads.
- Evaluated two mitogens: phorbol myristate acetate (PMA) with ionomycin and concanavalin A (ConA).
- Assessed the impact of stimulation strategies on T cell activation, expansion, transduction efficiency, CD4/CD8 T cell subset proportions, and regulatory T cell (Treg) prevalence.
Main Results:
- Plate-bound antibodies were significantly more effective than bead-bound antibodies for canine T cell stimulation.
- Plate-bound αCD3 clone CA17.6F9 combined with αCD28 clone 5B8, or with the mitogen PMA/ionomycin, yielded superior activation, expansion, and transduction.
- These optimal strategies also resulted in favorable T cell subset profiles, including CD4/CD8 ratios and Treg prevalence, potentially improving therapeutic outcomes.
Conclusions:
- Plate-bound antibody stimulation, particularly using specific αCD3/αCD28 combinations or PMA/ionomycin, is superior for canine T cell immunotherapy.
- Optimized T cell activation and transduction strategies can enhance the efficacy and safety of T cell-based cancer treatments in dogs.
- Findings provide a critical foundation for advancing canine T cell immunotherapy, with potential implications for human cancer therapies.
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