Vinsamlegast notið þetta auðkenni þegar þið vitnið til verksins eða tengið í það: http://hdl.handle.net/1946/44670
Chronic, improperly-resolved inflammation is involved in the pathology of many autoimmune and degenerative diseases, including asthma, arthritis, and cancer. Natural Killer (NK) cells are innate lymphocytes which have long been studied for their anti-viral and anti-cancer properties, as well as their promotion of inflammation. However, NK cells have also been shown to be critical for the resolution of inflammation. In addition, NK cell phenotype has been shown to be plastic and inducible in culture. This study determined the phenotypic changes NK cells undergo during treatment with pro-resolving and anti-inflammatory molecules to determine the inducibility of NK cells in vitro towards a pro-resolution phenotype.Peripheral blood NK cells were isolated from healthy donors and incubated with IL-15 for 18 h. The cells were then stimulated with resolution-inducing mediators: IL-10, TGF-β, and RvE1, inflammatory-inducing mediators: IL-2 and IL-12 (INF) along with IL-15, or with IL-15 only. Production of pro-resolving and pro-inflammatory mediators were determined by SimpleWestern (cell lysate, 6 h) and enzyme-linked immunosorbent assay (ELISA) (cell culture supernatant, 24 h). Expression of surface molecules was determined by flow cytometry (FC) after 6 h in culture. Induction of neutrophil apoptosis was measured by FC analysis of Annexin V expression on neutrophils after their co-culturing with RES- or INF-stimulated NK cells for 6 h. RES-treated NK cells produced more of the specialized pro-resolving mediators Annexin A1 and Lipoxin A4 than INF-treated NK cells. RES-treated NK cells secreted less of the pro-inflammatory mediators IFN-γ and GM-CSF than NK cells stimulated with INF. Cell surface expression of several activating receptors DNAM and 2B4 were decreased when NK cells were stimulated with RES compared with that when stimulated with INF. RES treatment, however, had no effect on NK cell induction of neutrophil apoptosis or other measures of cytotoxicity, namely granzyme B secretion and TRAIL and FasL expression.
The results presented in this project indicate that NK cells can be induced in vitro to possess pro-resolution phenotypic attributes. This provides support for the possible manipulation of NK cells in vitro for use in therapeutic intervention and the promotion of resolution of inflammation.
|JW thesis 2023.pdf