Current advances in experimental and computational approaches to enhance CAR T cell manufacturing protocols and improve clinical efficacy

Since the FDA’s approval of chimeric antigen receptor (CAR) T cells in 2017, significant improvements have been made in the design of chimeric antigen receptor constructs and in the manufacturing of CAR T cell therapies resulting in increased in vivo CAR T cell persistence and improved clinical outcome in certain hematological malignancies. Despite the remarkable clinical response seen in some patients, challenges remain in achieving durable long-term tumor-free survival, reducing therapy associated malignancies and toxicities, and expanding on the types of cancers that can be treated with this therapeutic modality. Careful analysis of the biological factors demarcating efficacious from suboptimal CAR T cell responses will be of paramount importance to address these shortcomings. With the ever-expanding toolbox of experimental approaches, single-cell technologies, and computational resources, there is renowned interest in discovering new ways to streamline the development and validation of new CAR T cell products. Better and more accurate prognostic and predictive models can be developed to help guide and inform clinical decision making by incorporating these approaches into translational and clinical workflows. In this review, we provide a brief overview of recent advancements in CAR T cell manufacturing and describe the strategies used to selectively expand specific phenotypic subsets. Additionally, we review experimental approaches to assess CAR T cell functionality and summarize current in silico methods which have the potential to improve CAR T cell manufacturing and predict clinical outcomes

Effect of Regulatory T Cells on the Humoral Immune Response to Borrelia Burgdorferi

Lyme borreliosis, caused by the bacteria Borrelia burgdorferi, is the most common tick-borne disease in the United States. Difficulties and delays in diagnosing can leave patients with long-term illness affecting multiple body systems. Therefore, understanding the immunomodulatory mechanisms affecting disease pathology will aid in the development of therapeutics and/or a safe, effective vaccine. In this thesis, we aimed to test the hypothesis that depletion of regulatory T cells amplifies the humoral response to Borrelia burgdorferi but leads to worsened clinical manifestations. To test this hypothesis, “depletion of regulatory T cell” (“DEREG”) BALB/c mice were depleted of Treg cells prior to infection with B. burgdorferi and tibiotarsal joint swelling, histopathology, and IgG titers were assessed. The Treg cell-depleted DEREG mice that were infected trended towards having the greatest pathology in their tibiotarsal joint swelling measurements and histopathology scores, lending partial support to our central hypothesis. Additionally, to lay the foundation for future studies on the regulation of the memory humoral response to B. burgdorferi in a DEREG BALB/c model, we tested various vaccine doses in mice and challenged them with a homologous strain of B. burgdorferi three weeks later. We found that it would take a dose of at least 1x106 heat-inactivated organisms to induce a protective response capable of inhibiting immune cell infiltration in the tibiotarsal joints of wild-type BALB/c mice. While further studies are needed to better define the role of Treg cells in the humoral and protective memory responses to B. burgdorferi infection, this study adds partial support to the hypothesis that Treg cell depletion prior to infection with B. burgdorferi increases the clinical manifestations of diseas