| 1. |
- Albertsson, Per, 1964, et al.
(författare)
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Differential locomotion of long- and short-term IL-2-activated murine natural killer cells in a model matrix environment.
- 2007
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Ingår i: Scandinavian journal of immunology. - : Wiley. - 0300-9475 .- 1365-3083. ; 66:4, s. 402-9
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Tidskriftsartikel (refereegranskat)abstract
- Tumour infiltration by activated natural killer (A-NK) cells is a pre-requisite for tumour eradication by adoptive NK cell transfer. Extravasated A-NK cells do not always succeed in reaching the crucial target cell conjugation. Therefore, we wished to study A-NK cell locomotion and interactions with melanoma cells in a matrix environment (Matrigel) by electron, confocal and fluorescence microscopy. Two distinct patterns of A-NK cell-mediated matrix disintegration were revealed during incubation of tumour cells and A-NK cells in Matrigel: (1) A-NK cells pre-cultured for 5 days altered the homogeneous texture of the Matrigel, an initial microporous appearance became a loose filamentous meshwork by 24 h. Matrix degrading protease inhibitors could not fully prevent this, but could delay the process; and (2) A-NK cells pre-cultured for 6 days or more, instead formed large excavations in the Matrigel leaving the remaining matrix less affected compared to the effects by the younger A-NK cells. By histochemical staining with Cupromeronic Blue, the excavations were shown to contain proteoglycan material. Protease inhibitors had no discernable effect on the development of the excavations. The conspicuous capacity of A-NK cells to disintegrate extracellular matrix and the formation of large excavations seems only partially to depend on matrix-degrading proteases. Formation of extracellular proteoglycan material is suggested to facilitate A-NK cell locomotion within a matrix environment.
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| 2. |
- Edsparr, Karin, 1979, et al.
(författare)
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Effects of IL-2 on MMP expression in freshly isolated human NK cells and the IL-2-independent NK cell line YT.
- 2010
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Ingår i: Journal of immunotherapy (Hagerstown, Md. : 1997). - 1537-4513. ; 33:5, s. 475-81
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Tidskriftsartikel (refereegranskat)abstract
- Interleukin-2 is an important activation factor for natural killer (NK) cells but its effect on NK cell matrix metalloproteinases (MMP) production and matrix degradation is less well investigated. We have used freshly isolated human NK cells and the IL-2-independent NK cell line, YT, to investigate the effects of IL-2 stimulation on NK cell invasion of Matrigel and on MMP expression and production. In YT cells, we found opposing early and late effects of IL-2 stimulation with an early (2 h) increase in MMP-9 protein level and enhanced migration in the Matrigel invasion assay and by 30 hours a decreased mRNA expression of MMP-2, MMP-9, MMP-13, MT3-MMP, and MT6-MMP. We also found a preculture period of 48 hours with IL-2 to negatively affect YT cell migration. We furthermore found that freshly isolated human NK cells Matrigel invasion was MMP-dependent and it increased in response to IL-2. Importantly, in freshly isolated human NK cells we did not see a downregulation of MMPs after 24 hours IL-2 stimulation, but instead a significant upregulation of MT6-MMP mRNA. Because of the cellular localisation of MT6-MMP, which ensures a focalized proteolytic activity, and its high expression compared with the other MMPs in freshly isolated human NK cells makes it of interest to study further.
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| 3. |
- Edsparr, Karin, 1979, et al.
(författare)
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Human NK cell lines migrate differentially in vitro related to matrix interaction and MMP expression.
- 2009
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Ingår i: Immunology and cell biology. - : Wiley. - 1440-1711 .- 0818-9641. ; 87:6, s. 489-95
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Tidskriftsartikel (refereegranskat)abstract
- Matrix metalloproteinases (MMPs) are thought to be of importance for the migratory ability of natural killer (NK) cells. Their expression and production may influence the amount of tumour-infiltrating NK cells and thereby any therapeutic capability. In this study, we sought to investigate the importance of MMPs for human NK cells' ability to degrade and migrate through the extracellular matrix (ECM). The two human NK cell lines, NK-92 and YT, migratory ability, MMP expression and production as well as their morphological appearance when cultured in the ECM equivalent Matrigel were analysed and compared. The quantitatively more migratory NK-92 cells were found to express invadopodia/podosomes at a significantly higher degree when cultured in Matrigel and gave rise to a general disintegration of the Matrigel. The NK-92 cells had a higher mRNA expression of MMP-2, -9, -13, MT1-, MT3- and MT6-MMP and a significantly higher production of MMP-9 compared to YT cells. These differences could explain the substantial functional difference observed between the two cell lines with respect to migratory capacity. In addition, the number of Matrigel invading NK-92 cells decreased significantly in the presence of the MMP inhibitor GM6001, demonstrating that MMPs have a critical function in their migration.
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| 4. |
- Edsparr, Karin, 1979, et al.
(författare)
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Matrix metalloproteinases in cytotoxic lymphocytes impact on tumour infiltration and immunomodulation.
- 2011
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Ingår i: Cancer microenvironment : official journal of the International Cancer Microenvironment Society. - : Springer Science and Business Media LLC. - 1875-2284. ; 4:3, s. 351-60
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Tidskriftsartikel (refereegranskat)abstract
- To efficiently combat solid tumours, endogenously or adoptively transferred cytotoxic T cells and natural killer (NK) cells, need to leave the vasculature, traverse the interstitium and ultimately infiltrate the tumour mass. During this locomotion and migration in the three dimensional environment many obstacles need to be overcome, one of which is the possible impediment of the extracellular matrix. The first and obvious one is the sub-endothelial basement membrane but the infiltrating cells will also meet other, both loose and tight, matrix structures that need to be overridden. Matrix metalloproteinases (MMPs) are believed to be one of the most important endoprotease families, with more than 25 members, which together have function on all known matrix components. This review summarizes what is known on synthesis, expression patterns and regulation of MMPs in cytotoxic lymphocytes and their possible role in the process of tumour infiltration. We also discuss different functions of MMPs as well as the possible use of other lymphocyte proteases for matrix degradation.
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| 5. |
- Edsparr, Karin, 1979
(författare)
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Migration of Natural Killer Cells. Matrix interaction, locomotion and regulation of matrix metalloproteinases (MMPs) by IL-2 and chemokines
- 2009
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Activated natural killer (NK) cells are effective anti-tumour cells. In order to reach target cells in a tumour mass they need to migrate in the extravascular space. The process of tumour localisation begins when the NK cell is attracted in the blood stream, and utilisation of matrix-degrading enzymes is crucial for passing of the basal membrane (BM) and for locomotion outside the vascular bed. NK cells express several members of the family of matrix-degrading enzymes, matrix metalloproteinases (MMPs). The aim of this study was to gain more knowledge on the regulation of NK cell migration that affects infiltration of the extracellular matrix (ECM) equivalent Matrigel. In specific; morphologically study NK cell locomotion in a matrix environment; identify the repertoire of MMPs expressed by freshly isolated human NK cells and the human NK cell lines YT and NK-92; explore the role of MMPs in NK cell migration and investigate the effect of IL-2 and chemokine stimulation as well as matrix (Matrigel) contact on NK cells’ migratory ability and MMP expression. IL-2-activated mouse A-NK cells cultured in Matrigel revealed two different patterns of matrix disintegration depending on their time in culture, and similar differences were found between two human NK cell lines. Younger (≤ 5 days) mouse A-NK and NK-92 cells gave rise to a general widespread matrix reorganisation, interpreted to be due to direct release of soluble matrix-degrading enzymes. Older (≥ 6 days) mouse A-NK and YT cells instead produced large excavations in the Matrigel. These cavities could be explained by a release of proteoglycan-rich material with matrix-dilating properties, combined with associated matrix-degrading proteases. The IL 2 independent NK cell line YT and freshly isolated human NK cells was used to investigate the effects of IL-2 stimulation on NK cell migration and MMP expression and production. IL-2 stimulation of the YT cell line demonstrated opposing effects related to the duration of stimulation. A rapid stimulatory response at about 2-4h on MMP production, and a later negative effect on MMP expression and MMP-9 production was seen after prolonged stimulation (≥ 24h). Both responses correspondingly affected the migratory ability. In freshly isolated NK cells, migration increased MMP-dependently in response to IL-2 and MT6-MMP expression increased. MMP-13, MT3- and MT6-MMP, previously not described in NK cells, was found to be expressed by freshly isolated human NK cells. While matrix (Matrigel) contact did not affect MMP expression in either the NK-92 or YT cell line, the chemokine CX3CL1 was found to increase NK-92 cells’ MMP-9 production significantly, but had no effect on their migration. These findings increase our understanding of how NK cell migration is regulated and provide one further step in the development of strategies to achieve greater number of tumour infiltrating NK cells.
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| 6. |
- Novotny, Ann, 1982, et al.
(författare)
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A pharmacological analysis of the cholinergic regulation of urokinase-type plasminogen activator secretion in the human colon cancer cell line, HT-29.
- 2010
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Ingår i: European journal of pharmacology. - : Elsevier BV. - 1879-0712 .- 0014-2999. ; 646:1-3, s. 22-30
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Tidskriftsartikel (refereegranskat)abstract
- Urokinase-type plasminogen activator (uPA) is an important factor for tumour cell invasion and metastasis. We recently showed that acetylcholine is an autocrine/paracrine growth factor for the human colon cancer cell line, HT-29, in part via the alpha7 subtype of the nicotinic acetylcholine receptors. In the current study, we investigated whether acetylcholine participates in the regulation of the protein expressions of also uPA and its receptor (uPAR) in the HT-29 cell line. Such were investigated by immunocytochemistry and Western blotting, and quantitation of uPA secretion was undertaken by ELISA. Stimulation of the cells for 24h with nicotine caused increased uPA secretion with peak effect (78% above the control) occurring at a nicotine concentration of 10nM. This effect was markedly inhibited by alpha-Bungarotoxin, thus showing the involvement of alpha7 nicotinic acetylcholine receptors. Basal uPA secretion was found to be partly dependent on ongoing activation of nicotinic receptors, suggesting tonic production of acetylcholine. Conversely, there was no cholinergic influence on the expression of uPAR. The current findings demonstrate novel aspects of receptor-mediated regulation of tumour metastatic potential via uPA secretion. This may suggest future pharmaceutical strategies in treatment of colorectal cancer.
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