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- Title
- ACTIVATION AND EXPANSION OF NATURAL KILLER CELLS FOR CANCER IMMUNOTHERAPY WITH EX21 EXOSOMES.
- Creator
-
Khederzadeh, Sara, Copik, Alicja, University of Central Florida
- Abstract / Description
-
In the field of cancer immunotherapy, NK cells are recognized for their ability to provide a form of innate immunity against tumor cells. However, the average abundance of NK cells in the blood can be as low as 5% of the total lymphocyte population. As a result, it has been a focus to find novel therapies to expand NK cells in vitro while subsequently enhancing the cytotoxicity of these cells. Previously-defined methods include the minimal expansion of NK cells with high levels of cytokines...
Show moreIn the field of cancer immunotherapy, NK cells are recognized for their ability to provide a form of innate immunity against tumor cells. However, the average abundance of NK cells in the blood can be as low as 5% of the total lymphocyte population. As a result, it has been a focus to find novel therapies to expand NK cells in vitro while subsequently enhancing the cytotoxicity of these cells. Previously-defined methods include the minimal expansion of NK cells with high levels of cytokines such as IL-2 and IL-15, as well as co-culturing NK cells with feeder cell populations that are genetically modified to express NK-stimulating factors. Another method involves the use of artificially-derived plasma membrane nanoparticles (PM21) that express membrane-bound IL-21 (mb21) to successfully expand NK cells by a factor of 103 in 14 days. Exosomes, which are cell-derived vesicles naturally secreted by cancer cells, may reveal a novel way to expand NK cells and enhance their cytotoxicity by taking advantage of the exchange of genetic information within the tumor microenvironment. To test this hypothesis, NK cells have been cultured with varying concentrations of exosomes derived from modified K562-mb21-41BBl (a chronic myelogenous leukemia cell line) and shown to achieve 200-fold expansion of NK cells from other PBMCs in 14 days, a growth comparable to that of PM-21 particles. In vitro assays as well as co-culturing with various tumor cell lines will determine the cytotoxicity of these expanded cells. Potentially, exosomes may be applied as an in vivo therapy for NK cell expansion.
Show less - Date Issued
- 2017
- Identifier
- CFH2000262, ucf:45963
- Format
- Document (PDF)
- PURL
- http://purl.flvc.org/ucf/fd/CFH2000262
- Title
- DEVELOPMENT OF METHODS TO MODULATE NATURAL KILLER CELLS.
- Creator
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Shaver, Kari A, Copik, Alicja, University of Central Florida
- Abstract / Description
-
Natural Killer (NK) cell based immunotherapies have demonstrated success against malignancies and hematological cancers. However, tumors have developed mechanisms to evade detection by and suppress the immune system, commonly through altering the expression of cell-surface proteins. Overexpression of human leukocyte antigen-E (HLA-E), which binds to the inhibitory NKG2A on NK cells, protects malignant cells from lysis. Downregulating the NKG2A receptor on NK cells should release NK cell...
Show moreNatural Killer (NK) cell based immunotherapies have demonstrated success against malignancies and hematological cancers. However, tumors have developed mechanisms to evade detection by and suppress the immune system, commonly through altering the expression of cell-surface proteins. Overexpression of human leukocyte antigen-E (HLA-E), which binds to the inhibitory NKG2A on NK cells, protects malignant cells from lysis. Downregulating the NKG2A receptor on NK cells should release NK cell inhibition, but proves challenging as NK cells are difficult to transfect and no good methods currently exist. This project is designed to investigate the use of exosomes - small vesicles and natural carriers of regulatory microRNAs (miRNAs) and proteins that are shed from cells - as delivery vehicles for small RNAs (sRNAs) to immune cells. Exosomes are biologically compatible, immunologically inert, and interact with target cells through receptor-ligand interactions, allowing for targeted delivery of cargo. Exosomes loaded with shRNA against NKG2A were cultured in vitro with NK cells. Delivery success was assessed by monitoring NKG2A receptor expression on NK cells through flow cytometry. This research will provide valuable information that will likely impact the delivery of RNA therapeutics and unlock the full cytotoxic potential of NK immunotherapy.
Show less - Date Issued
- 2018
- Identifier
- CFH2000455, ucf:45720
- Format
- Document (PDF)
- PURL
- http://purl.flvc.org/ucf/fd/CFH2000455
- Title
- MULTIPLE ASPECTS OF NATURAL KILLER CELL EXPANSION IN RELEVANCE TO IMMUNOTHERAPY FOR HEMATOLOGIC MALIGNANCIES.
- Creator
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Colosimo, Dominic, Borgon, Robert, University of Central Florida
- Abstract / Description
-
Natural Killer (NK) cells are a subset of lymphocytes that regulate adaptive immune responses and utilize "missing self" recognition to activate anti-tumor and anti-viral cytotoxicity. Clinical research, as well as murine and ex vivo models, have shown that a variety of NK cell applications have proven useful as immunotherapeutic treatments for patients with hematologic malignancies. However, the selective expansion of NK cells to yield relevant amounts of these lymphocytes has been a major...
Show moreNatural Killer (NK) cells are a subset of lymphocytes that regulate adaptive immune responses and utilize "missing self" recognition to activate anti-tumor and anti-viral cytotoxicity. Clinical research, as well as murine and ex vivo models, have shown that a variety of NK cell applications have proven useful as immunotherapeutic treatments for patients with hematologic malignancies. However, the selective expansion of NK cells to yield relevant amounts of these lymphocytes has been a major hurdle in the development of methods for clinical therapeutic use. Here, we demonstrate a novel ex vivo expansion method utilizing k562 leukemic cell lines and soluble cytokines as well as a novel method utilizing isolated plasma membranes of genetically engineered tumor cell lines that could be of relevance to in vivo NK cell expansion. Also, the ligand expression by canonical feeder cell lines used for NK cell expansion and our isolated plasma membranes were compared via ligand quantification by western blot quantification of 4-1BB ligand. In an adjunct study, we sought to better characterize these expansion environments by investigating the glucose metabolism of NK cells using fluorescent glucose analog 2-(N-(7-Nitrobenz-2-oxa-1, 3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) and the glycolysis inhibitor 2-Deoxy-D-Glucose (2-DG).
Show less - Date Issued
- 2012
- Identifier
- CFH0004252, ucf:44917
- Format
- Document (PDF)
- PURL
- http://purl.flvc.org/ucf/fd/CFH0004252
