A Microfluidics-Based Approach For Isolation of Antigen-Specific Cd8+ T Cells

<p> Cancer is a global epidemic: there are predicted to be 200 million new cases this year alone. Almost a quarter of all cancer-related deaths are caused by lung cancer, for which 5-year survival rates are just above 20%. 85% of lung cancer diagnoses are classified as non-small cell lung cancer (NSCLC) for which 5-year survival rates in metastatic disease are less than 10%. Early detection and targeted therapies have improved prognoses, yet relapse is still common among patients.</p> <p> Immunotherapies that leverage tumor-specific CD8⁺cytotoxic T cells have shown great promise for the treatment of NSCLC. However, although highly promising, the success of these therapies has largely been hampered by the challenge of isolating tumor-specific CD8⁺ T cells from a bulk tumor-infiltrating lymphocyte (TIL) population, which are exceedingly rare. Current methods rely on the identification of tumor-specific antigens, which remains flawed. Current methods to isolate tumor-specific CD8⁺ T cells heavily rely on inaccurate antigen prediction models which require laborious functional validation in lab and make large-scale application infeasible.</p> <p> The objective of this study is to develop a method which <em>rapidly </em>and <em>specifically </em>isolates antigen-specific CD8⁺ T cells that bypasses the need for prior antigen identification. In this work, I optimized a novel microfluidics method, “ATTACH” (Assessment of T cells Tethered to Antigen Class I and II Histocompatibility) within the model Ovalbumin (OVA) antigen system. Using a <em>de facto </em>pool of peptide-loaded MHC class I molecules on the surface of target cells, ATTACH enriches for antigen-specific CD8⁺ T cells based on binding avidity to cognate antigens. Here, I demonstrate that ATTACH specifically enriched for OVA-specific OT-I CD8+ T cells from both bulk splenocyte and heterogeneous CD8⁺T cell populations. Importantly, enriched antigen-specific CD8⁺ T cell populations exhibited significantly greater antigen-specific effector function than both the bulk input and eluted non-specific CD8⁺ population.<strong></strong></p>

Name thesis:degree_name

Masters of Science (MS)

Level thesis:degree_level

Thesis (MS)

Year dc:date.available

2022

Authors dc:creator

• Frank, Meredith
• <p>0000-0002-3658-0057</p>

Contributors dc:contributor

• Alexandre Reuben
• John Heymach
• Pamela Wenzel

• Immunology
• CD8 T cell
• T cell
• NSCLC
• Cancer
• Neoantigen
• Microfluidics
• antigen-specific T cells
• Immunopathology
• Laboratory and Basic Science Research
• Translational Medical Research

Repository record dc:identifier

https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1210

OAI identifier oai:identifier

oai:digitalcommons.library.tmc.edu:utgsbs_dissertations-2267