Molecular mechanisms governing the identity and clinical functionality of type 1 regulatory t cells
Abstract/Contents
- Abstract
- Type 1 regulatory T cells (Tr1) are a peripherally derived regulatory T cell subset that confer antigen-specific tolerance in vivo. While Tr1 cells possess potent regulatory capabilities, they represent a rare T cell population. Therefore, the successful implementation of Tr1 based cell therapies will require simultaneous efforts to develop efficient in vitro induced Tr1 (iTr1) methods while uncovering basic Tr1 biology. One iTr1 method is the enforced expression of the IL-10 gene in CD4+ T cells, referred to here as LV-10 cells. LV-10 cells can dampen harmful immune responses while exhibiting myeloid-specific cytotoxicity, thus bringing forth a unique opportunity to concurrently reduce graft vs host disease after allogeneic hematopoietic stem cell transplantation for myeloid cancers and promote graft versus leukemia. Two major areas that are explored in this thesis are the identification of 1) transcription factors (TF) that control Tr1 identity and 2) protein interactions between leukemic blasts and LV-10 cells that impact cytotoxicity. The master TFs driving human Tr1 development are still unknown. We conducted the first transcriptome analysis on de novo human Tr1 by RNA sequencing. Utilizing a variety of tools such as gene knockout, gene overexpression, and in vitro differentiation, we have characterized the role of BHLHE40 in both primary human CD4+ T cells and naïve CD4+ T cells. Ultimately, identifying Tr1-specific TFs will improve our ability to detect, isolate, and manipulate endogenous Tr1, and design efficient protocols to generate iTr1 for cell therapy. LV-10 cells are an attractive iTr1 cell product because they allow for the efficient conversion of Tr1-like cells that secret high IL-10 while exhibiting myeloid cytotoxicity. We investigated the extent of their anti-leukemic effect on primary pediatric acute myeloid leukemia (pAML). We tested 23 pAML and found that pAML exhibited a range of sensitivities to LV-10 killing: sensitive, intermediate-resistant, and resistant. After comparing the immune phenotypes of pAML via flow cytometry and the transcriptome via RNA-sequencing, we determined that sensitive and resistant pAML are molecularly distinct. We specifically identified that resistant pAML express high levels of CD200 and after co-culturing CD200 overexpressing sensitive cell lines with LV-10 cells, we observed a reduction in LV-10 degranulation. Altogether, LV-10 cells represent a promising clinical use for pAML, though further characterization is needed to optimize their potential clinical usefulness.
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 | 2020; ©2020 |
| Publication date | 2020; 2020 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Uyeda, Molly Kathryn Javier |
|---|---|
| Degree supervisor | Roncarolo, Maria-Grazia |
| Thesis advisor | Roncarolo, Maria-Grazia |
| Thesis advisor | Cleary, Michael L |
| Thesis advisor | Nakauchi, Hiromitsu, 1952- |
| Thesis advisor | Shizuru, Judith Anne |
| Degree committee member | Cleary, Michael L |
| Degree committee member | Nakauchi, Hiromitsu, 1952- |
| Degree committee member | Shizuru, Judith Anne |
| Associated with | Stanford University, Department of Stem Cell Biology and Regenerative Medicine |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Molly Kathryn Javier Uyeda. |
|---|---|
| Note | Submitted to the Department of Stem Cell Biology and Regenerative Medicine. |
| Thesis | Thesis Ph.D. Stanford University 2020. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Molly Kathryn Javier Uyeda
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