Defining natural killer cell responses to HIV
Abstract/Contents
- Abstract
- Human immunodeficiency virus (HIV) remains a major global public health issue, and in the continued absence of a cure or vaccine, novel treatment and prevention strategies are needed to combat the ongoing epidemic. Many lines of evidence have suggested that natural killer (NK) cells, effector cells of the innate immune system that rapidly respond to virus-infected and malignantly transformed cells, can contribute to protection against HIV acquisition and disease progression. Here provide a characterization of methods of NK cell targeting of HIV, encompassing both natural receptor-ligand interactions in the setting of in vitro infection as well as in HIV-infected individuals, as well as via the use of bispecific antibodies to specifically target NK cells to HIV-infected cells. We first optimize an in vitro co-culture system to detect HIV-specific NK cell responses to autologous infected cells. We then use a combination of single cell transcriptomics and mass cytometry to characterize the features of NK cells in these co-cultures. Using single cell RNA-seq, we find a dominant interferon-driven gene signature in NK cells responding to HIV-infected cells. In addition, by mass cytometry profiling of NK cell phenotype and function, paired with profiling of ligand expression on infected CD4 T cells, we identify candidate receptor-ligand interactions (NKG2D/MIC-A/B and ULBP-1/2/5/6, and NKp30-B7-H6) involved in recognition mechanisms, which we then validate using CRISPR knockout of the corresponding ligands on target cells. We thus identify mechanisms of NK cell activation and response against HIV. We then study whether HIV-infected individuals treated with antiretroviral therapy (ART) possess alterations in NK cells that may make them more responsive to a second, in vitro exposure to HIV. We once again use mass cytometry to profile NK cell phenotype and function in both treated, HIV-infected individuals and healthy controls and find that, while there is a shift in the pattern of expression of NK cell receptors including NKp30 and CD2, the overall phenotype and frequency of the subset of cells that respond to in vitro HIV stimulation is not changed. We thus find that while the NK cell repertoire is altered in treated, HIV-infected individuals, it does not become more HIV-specific. Lastly, we develop and test a bispecific antibody that targets HIV gp41 and the NK cell activating receptor CD16. We show that this bispecific antibody has increased affinity and avidity to CD16, and is thus able to robustly enhance NK cell targeting of infected cells. This provides a means for controlled targeting of NK cells to infected cells. Overall, we identify multiple mechanisms that may hold promise to improve NK cell activity against HIV-infected cells. We discuss the extension of these findings in improving NK cell-mediated immunotherapy in HIV infection
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 | Zhao, Qi |
|---|---|
| Degree supervisor | Blish, Catherine |
| Thesis advisor | Blish, Catherine |
| Thesis advisor | Kim, Peter, 1958- |
| Thesis advisor | Sarnow, P. (Peter) |
| Thesis advisor | Sunwoo, John B |
| Degree committee member | Kim, Peter, 1958- |
| Degree committee member | Sarnow, P. (Peter) |
| Degree committee member | Sunwoo, John B |
| Associated with | Stanford University, Department of Immunology. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Nancy Qi Zhao |
|---|---|
| Note | Submitted to the Department of Immunology |
| Thesis | Thesis Ph.D. Stanford University 2020 |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Qi Zhao
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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