Högupplöst bildtagning av cytoskelett protein och och pkc-o i NK-celler stimulerade av synapsformade mönster
Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
High resolution imaging of cytoskeletal proteins and pkc-o in natural killer cells stimulated by synaptic patterns (English)
Natural killer cells are lymphocytes vital to the immune system by eliminating abnormal cells by the release of cytolytic proteins. Upon target cell encounter an immune synapse is formed at the interface between the NK-cell and the target cell, in this synapse interacting receptors on the NK cell form supramolecular activation clusters followed by a range of intracellular events including actin reorganization and MTOC polarization. So far little has been done to resolve how receptor clustering and intracellular events depend on the spatial distribution of target cellligands. To investigate how activation by differently distributed ligands affect NKcells we used microcontact printing to pattern antibodies to the NK cell receptorsCD16 and LFA-1 in torus or dot micro-scale patterns. Our findings showed that LFA-1 activation triggered an elongated migratory morphology in NK cells, addition of CD16 stimulus elicited a round morphology with the NK cells covering the print. NKcells stimulated by CD16 polarized MTOC further relative to LFA-1 stimulus alone, however stimuli by LFA-1, CD16 or both polarized the MTOC towards the surface regardless of printed pattern. While polarization was not dependent on the printed pattern, we found that positioning of the MTOC in the surface plane was dependent on received stimuli and ligand distribution. PKC-ɵ was in our experiment distributed around the NK cell body and was not affected by choice of protein and pattern. Finally we discovered that small actin holes emerged in NK cells interacting with thecombination of CD16 and LFA-1 regardless of pattern design.
Place, publisher, year, edition, pages
immune cell, natural killer cell, confocal microscopy, immune synapse, super-resolution microscopy, receptor
Engineering and Technology
IdentifiersURN: urn:nbn:se:kth:diva-163704OAI: oai:DiVA.org:kth-163704DiVA: diva2:802029