Date of Award
Doctor of Philosophy (PhD)
Biotechnology Science and Engineering
T cells., Biomedical engineering.
T cells are crucial components of the adaptive immune system and are known for secreting various cytokines, which induce the proliferation of many immune cells, including T cells themselves. A subset of T cells, called the CD8+ T cells, are known for their ability to kill infected cells and cancer cells. Naturally, the potential in using T cells for immunotherapy of cancer is tremendous. For this purpose, however, it is often required to engineer T cells specifically to treat a particular type of cancer. For example, the FDA-approved immunotherapies, KymriahTM and YescartaTM, which come under the adoptive T cell therapy, require genetic engineering and in vitro expansion of T cells. Currently, viral vectors are being used to introduce the transgenes encoding the chimeric antigen receptor (CAR) to patient-derived T cells in vitro. Due to the safety concerns and high manufacturing costs associated with viral transduction, non-viral vector-mediated T cell-specific transfections are being developed. Besides, the patient’s natural Antigen-presenting cells (APCs), such as the dendritic cells, may not offer consistent activation profiles. Thus, the function of CAR-T cells must be modulated with artificial antigen-specific APCs for more specific and controlled activation profiles, whose manufacturing and maintenance are easier and less expensive. Therefore, it is imperative that we understand T cell biology, improve the engineering and functional modulation strategies to advance the field of T cell therapy. Here, we suggest how to improve the non-viral vector-mediated transfection efficiency in Jurkat cells (a human leukemia T cell line) by using calcium ions; how to express and purify N-terminal Cysteine containing mouse Interleukin-2 (IL-2) at the laboratory scale to enable site-specific bioconjugation of IL-2 with foreign materials; how to associate and intracellularly deliver the IL-2-functionalized nanocargoes up to a certain size-limit to the murine primary T Lymphocytes via the IL-2 receptor (IL-2R)-mediated endocytosis; and finally, how to optimize the manufacture of human HER2- and PD-L1-coated iron-oxide microparticles to functionally modulate and test the efficacies of the HER2 CAR-T cells.
Ayyadevara, Venkata Sesha Sai Abhinav, "Engineering and functional modification strategies for T lymphocytes using therapeutic macromolecules, polymers, micro and nanoparticles" (2021). Dissertations. 232.