Comprehensive Approach for Identifying the T Cell Subset Origin of CD3 and CD28 Antibody–Activated Chimeric Antigen Receptor–Modified T Cells
| Autor: | Petra Reinke, Lisa Rollins, Michael Schmueck-Henneresse, Natalia Lapteva, Sandhya Sharma, Gianpietro Dotti, Thomas Shum, Maksim Mamonkin, Bilal Omer, Haruko Tashiro, Cliona M. Rooney, Hans-Dieter Volk |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Cytotoxicity
Immunologic 0301 basic medicine CD3 Complex Naive T cell T cell Immunology Receptors Antigen T-Cell Streptamer Biology Lymphocyte Activation Article Immunophenotyping 03 medical and health sciences Interleukin 21 CD28 Antigens T-Lymphocyte Subsets medicine Humans Immunology and Allergy Cytotoxic T cell IL-2 receptor Antigen-presenting cell Interleukin-15 Interleukin-7 Cell Differentiation Flow Cytometry Natural killer T cell Cell biology 030104 developmental biology medicine.anatomical_structure Interleukin-2 Leukocyte Common Antigens |
| Zdroj: | The Journal of Immunology. 199:348-362 |
| ISSN: | 1550-6606 0022-1767 |
| DOI: | 10.4049/jimmunol.1601494 |
| Popis: | The outcome of therapy with chimeric Ag receptor (CAR)-modified T cells is strongly influenced by the subset origin of the infused T cells. However, because polyclonally activated T cells acquire a largely CD45RO+CCR7− effector memory phenotype after expansion, regardless of subset origin, it is impossible to know which subsets contribute to the final T cell product. To determine the contribution of naive T cell, memory stem T cell, central memory T cell, effector memory T cell, and terminally differentiated effector T cell populations to the CD3 and CD28–activated CAR-modified T cells that we use for therapy, we followed the fate and function of individually sorted CAR-modified T cell subsets after activation with CD3 and CD28 Abs (CD3/28), transduction and culture alone, or after reconstitution into the relevant subset-depleted population. We show that all subsets are sensitive to CAR transduction, and each developed a distinct T cell functional profile during culture. Naive-derived T cells showed the greatest rate of proliferation but had more limited effector functions and reduced killing compared with memory-derived populations. When cultured in the presence of memory T cells, naive-derived T cells show increased differentiation, reduced effector cytokine production, and a reduced reproliferative response to CAR stimulation. CD3/28-activated T cells expanded in IL-7 and IL-15 produced greater expansion of memory stem T cells and central memory T cell–derived T cells compared with IL-2. Our strategy provides a powerful tool to elucidate the characteristics of CAR-modified T cells, regardless of the protocol used for expansion, reveals the functional properties of each expanded T cell subset, and paves the way for a more detailed evaluation of the effects of manufacturing changes on the subset contribution to in vitro–expanded T cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|