Author ORCID Identifier

0000-0002-3658-0057

Date of Graduation

8-2022

Document Type

Thesis (MS)

Program Affiliation

Biomedical Sciences

Degree Name

Masters of Science (MS)

Advisor/Committee Chair

Alexandre Reuben

Committee Member

John Heymach

Committee Member

Pamela Wenzel

Committee Member

Greg Lizee

Committee Member

Matthew Gubin

Committee Member

Askar Akimzhanov

Abstract

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.

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.

The objective of this study is to develop a method which rapidly and specifically 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 de facto 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.

Keywords

Immunology, CD8 T cell, T cell, NSCLC, Cancer, Neoantigen, Microfluidics, antigen-specific T cells

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