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.
","abstract_html":"<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<sup>+ </sup>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<sup>+</sup> 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<sup>+</sup> 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<sup>+</sup> 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<sup>+</sup> 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<sup>+ </sup>T cell populations. Importantly, enriched antigen-specific CD8<sup>+</sup> T cell populations exhibited significantly greater antigen-specific effector function than both the bulk input and eluted non-specific CD8<sup>+</sup> population.<strong></strong></p>","abstract_has_math":false,"creators":["Frank, Meredith","0000-0002-3658-0057
"],"institution":null,"degree_name":"Masters of Science (MS)","degree_level":"Thesis (MS)","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Alexandre Reuben","John Heymach","Pamela Wenzel"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2022,"date_issued":"2022-08-01T07:00:00Z","date_published":"2022-08-01T07:00:00Z","updated_at":"2026-07-24T05:50:09Z","subjects":["Immunology","CD8 T cell","T cell","NSCLC","Cancer","Neoantigen","Microfluidics","antigen-specific T cells","Immunopathology","Laboratory and Basic Science Research","Translational Medical Research"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1210","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Alexandre Reuben","John Heymach","Pamela Wenzel"]},{"key":"dc:creator","label":"Author","values":["Frank, Meredith","0000-0002-3658-0057
"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2023-01-31T08:00:00Z"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis (MS)"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Masters of Science (MS)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Immunology","CD8 T cell","T cell","NSCLC","Cancer","Neoantigen","Microfluidics","antigen-specific T cells","Immunopathology","Laboratory and Basic Science Research","Translational Medical Research"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1210"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["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.
"]},{"key":"dc:title","label":"Title","values":["A Microfluidics-Based Approach For Isolation of Antigen-Specific Cd8+ T Cells"]}]}],"canonical_facts":{"dc:contributor":["Alexandre Reuben","John Heymach","Pamela Wenzel"],"dc:creator":["Frank, Meredith","0000-0002-3658-0057
"],"dc:date.available":["2023-01-31T08:00:00Z"],"dc:description.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.
"],"dc:identifier":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1210"],"dc:subject":["Immunology","CD8 T cell","T cell","NSCLC","Cancer","Neoantigen","Microfluidics","antigen-specific T cells","Immunopathology","Laboratory and Basic Science Research","Translational Medical Research"],"dc:title":["A Microfluidics-Based Approach For Isolation of Antigen-Specific Cd8+ T Cells"],"thesis:degree_level":["Thesis (MS)"],"thesis:degree_name":["Masters of Science (MS)"]},"updated_at":"2026-07-24T05:50:09Z"}