Selecting and sequencing clonally expanded CD8+ T cells
Abstract/Contents
- Abstract
- The immune system is tasked with detecting and neutralizing infectious organisms to maintain homeostasis and prevent disease. This defense is managed by complex coordination of many cell types. CD8+ T cells are responsible for recognizing infected cells and killing them. To permit the recognition of antigens, T cells generate a vast diversity of T cell receptor (TCR) sequences. Upon binding of the TCR to an antigen-MHC complex, T cells clonally expand to establish an immune response. To study antigen-specific T cell clonality, we have developed a method that allows selection of rare cells, based on RNA expression, prior to in-depth scRNA-seq (named SELECT-Seq). We applied SELECT-Seq to collect both TCR sequences and then transcriptomes from single cells of peripheral blood lymphocytes activated by a Mycobacterium tuberculosis (Mtb) lysate. TCR sequence analysis allowed us to preferentially select expanded conventional CD8+ T cells as well as invariant natural killer T (iNKT) cells and mucosal-associated invariant T (MAIT) cells. The iNKT and MAIT cells have a highly similar transcriptional pattern, indicating that they carry out similar immunological functions, and differ considerably from conventional CD8+ T cells. While there is no relationship between expression profiles and clonal expansion in iNKT or MAIT cells, highly expanded conventional CD8+ T cells downregulate the interleukin-2 (IL-2) receptor alpha (IL2RA, or CD25) protein and show signs of senescence. This suggests inherent limits to clonal expansion that act to diversify the T cell response repertoire.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2019; ©2019 |
| Publication date | 2019; 2019 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Sikora, Michael Joseph |
|---|---|
| Degree supervisor | Steinmetz, Lars |
| Thesis advisor | Steinmetz, Lars |
| Thesis advisor | Davis, Ronald W. (Ronald Wayne), 1941- |
| Thesis advisor | Montgomery, Stephen, 1979- |
| Thesis advisor | Snyder, Michael, Ph. D |
| Degree committee member | Davis, Ronald W. (Ronald Wayne), 1941- |
| Degree committee member | Montgomery, Stephen, 1979- |
| Degree committee member | Snyder, Michael, Ph. D |
| Associated with | Stanford University, Department of Genetics. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Michael Joseph Sikora. |
|---|---|
| Note | Submitted to the Department of Genetics. |
| Thesis | Thesis Ph.D. Stanford University 2019. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2019 by Michael Joseph Sikora
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