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- Liu, Lisa L., et al.
(författare)
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Ex Vivo Expanded Adaptive NK Cells Effectively Kill Primary Acute Lymphoblastic Leukemia Cells
- 2017
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Ingår i: CANCER IMMUNOLOGY RESEARCH. - 2326-6066 .- 2326-6074. ; 5:8, s. 654-665
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Tidskriftsartikel (refereegranskat)abstract
- Manipulation of human natural killer (NK) cell repertoires promises more effective strategies for NK cell-based cancer immunotherapy. A subset of highly differentiated NK cells, termed adaptive NK cells, expands naturally in vivo in response to human cytomegalovirus (HCMV) infection, carries unique repertoires of inhibitory killer cell immunoglobulin-like receptors (KIR), and displays strong cytotoxicity against tumor cells. Here, we established a robust and scalable protocol for ex vivo generation and expansion of adaptive NK cells for cell therapy against pediatric acute lymphoblastic leukemia (ALL). Culture of polyclonal NK cells together with feeder cells expressing HLA-E, the ligand for the activating NKG2C receptor, led to selective expansion of adaptive NK cells with enhanced allor-eactivity against HLA-mismatched targets. The ex vivo expanded adaptive NK cells gradually obtained a more differentiated phenotype and were specific and highly efficient killers of allogeneic pediatric T-and precursor B-cell acute lymphoblastic leukemia (ALL) blasts, previously shown to be refractory to killing by autologous NK cells and the NK-cell line NK92 currently in clinical testing. Selective expansion of NK cells that express one single inhibitory KIR for self-HLA class I would allow exploitation of the full potential of NK-cell alloreactivity in cancer immunotherapy. In summary, our data suggest that adaptive NK cells may hold utility for therapy of refractory ALL, either as a bridge to transplant or for patients that lack stem cell donors.
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| 3. |
- Pfefferle, Aline
(författare)
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Dynamics of natural killer cell homeostasis : implications for cell-based cancer immunotherapy
- 2019
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Natural killer (NK) cells comprise a central role within the innate immune system, eliminating virally infected, foreign and transformed cells through their natural cytotoxic capacity. Release of their cytotoxic granules is tightly controlled through the balance of a large repertoire of inhibitory and activating receptors, and it is the unique combination of these receptors on individual cells that confers them their immense diversity both in phenotype and functionality. This thesis aimed to investigate the mechanisms sustaining NK cell homeostasis with the aim of translating these findings into more efficient NK cell-based immunotherapies against cancer. In paper I, we set out to define a transcriptional timeline for NK cell differentiation through the use of single-cell RNA sequencing of unique differentiation subsets ranging from CD56bright to adaptive NKG2C+CD56dim NK cells. Transcriptional differentiation was concentrated within the surprisingly diverse CD56bright subset which gradually transitioned into CD56dim NK cells before terminal differentiation into adaptive CD56dim NK cells. The vastly diverse yet unique NK cell repertoire within an individual is surprisingly stable over time considering the constant renewal of these cells at steady state. In paper II, we performed an in-depth analysis of homeostatic proliferation in human NK cells. We identified a high degree of intra-lineage plasticity combined with transcriptional reprogramming associated with the acquired phenotype as the underlying mechanisms maintaining repertoire stability at steady state. In paper III, we examined the role of NK cells in a setting of perturbed homeostasis, namely patients with high-risk myelodysplastic syndrome undergoing immunomodulatory treatment with 5-azacytidine. We identified a role for 5-azacytidine in modifying the global NK cell repertoire, as uptake of the drug by proliferating NK cells resulted in increased expression of killer cell immunoglobulin-like receptors (KIR) and improved functionality. In paper IV we identified a dose-dependent cytokine addiction in IL-15 expanded NK cells, leading to the induction of apoptosis upon cytokine withdrawal. A proliferation-dependent induction of the short splice variant of BIM, combined with an altered BCL-2/BIM ratio resulted in sensitization to cell death post withdrawal. This thesis provides new insights into the dynamic nature of NK cell homeostasis, from understanding NK cell differentiation at the transcriptional level to perturbations after cytokine stimulation and immunomodulatory therapies.
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- Wennerberg, Erik, et al.
(författare)
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Human anaplastic thyroid carcinoma cells are sensitive to NK cell-mediated lysis via ULBP2/5/6 and chemoattract NK cells
- 2014
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Ingår i: Clinical Cancer Research. - 1078-0432. ; 20:22, s. 5733-5744
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Tidskriftsartikel (refereegranskat)abstract
- PURPOSE: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive forms of cancer with no curative therapies available. To date, strategies to target ATC by immunotherapy have not been evaluated. We investigated whether ATC would be a suitable target for natural killer (NK) cell-based immunotherapy.EXPERIMENTAL DESIGN: We first established seven new cell lines from ATC tumors, three from papillary thyroid carcinoma tumors and analyzed them together with eight additional ATC cell lines. Cells were analyzed for sensitivity to lysis by NK cells and their ability to chemoattract and regulate the activity of NK cells. In addition, fresh tumor samples and peripheral blood from six patients with ATC were analyzed for NK cell infiltration and phenotype.RESULTS: We observed that ATC cell lines are sensitive to lysis by ex vivo expanded NK cells and that the lysis was abrogated upon blockade of NKG2D. Sensitivity of thyroid cancer cell lines to NK cell-mediated lysis correlated with surface expression of UL16-binding protein 2 on tumor cells. Moreover, ATC cell lines produced high levels of CXCL10 and stimulated migration of expanded NK cells and ATC tumors were enriched for NK cells expressing the cognate chemokine receptor CXCR3. However, compared with NK cells in peripheral blood, ATC tumor-derived NK cells displayed a suppressed phenotype with a downregulated expression of NKG2D. In vitro, suppression of NK cell-mediated lysis and NKG2D expression by ATC cells was restored upon neutralization of prostaglandin-E2.CONCLUSIONS: ATC cell lines are sensitive to NK cell-mediated lysis via ULBP2/5/6 and chemoattract CXCR3-positive NK cells. Patients with ATC may benefit from NK cell-based immunotherapy.
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