Proteolytic Activation of the Cytotoxic Phenotype during Human NK Cell Development

Meade, Josephine L.; Wilson, Erica; Holmes, Tim D.; Wynter, Erika A. de; Brett, Peter; Straszynski, Liz; Ballard, Paul A. S.; Trapani, Joseph A.; McDermott, Michael F.; Cook, Graham P.

doi:10.4049/jimmunol.0713829

Cited by 19 publications

(21 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…For IL-2 stimulation, NK cells were cultured for 5-7 days in 50 units/ml IL-2 (R&D systems). NK cell mediated killing of tumour cells and granule exocytosis assays were performed as we have described previously [24][25][26], including after siRNA transfection of target cells [26,27]. T cells restricted to the HLA-A2…”

Section: Preparation and Functional Analysis Of Nk Cells And Mart-1 Smentioning

confidence: 99%

Blocking oncogenic RAS enhances tumour cell surface MHC class I expression but does not alter susceptibility to cytotoxic lymphocytes

El-Jawhari

El-Sherbiny

Scott

et al. 2014

Molecular Immunology

Self Cite

View full text Add to dashboard Cite

Section: Preparation and Functional Analysis Of Nk Cells And Mart-1 Smentioning

confidence: 99%

Blocking oncogenic RAS enhances tumour cell surface MHC class I expression but does not alter susceptibility to cytotoxic lymphocytes

El-Jawhari

El-Sherbiny

Scott

et al. 2014

Molecular Immunology

Self Cite

View full text Add to dashboard Cite

“…To compare NK cell degranulation in response to monolayers or MCTS, an equal surface area of target cells was calculated for MCTS using an eye-piece graticule to measure MCTS radius (r) and the surface area calculated using the formula: area = 4(pr 2 ). The degranulation assays were performed as previously described (14,15). For gross analysis of spheroid death, similar sized spheroids were individually placed in separate wells of an agarcoated 96-well plate.…”

Section: Cell Culture and Transfectionmentioning

confidence: 99%

“…However, we have defined the threshold according to the susceptibility of target cells to a polyclonal population of NK cells exhibiting a normal distribution of killing activity; weak effector cells may spare target cells with subthreshold MHC class I expression levels, but these targets will eventually be detected and killed by strongly responsive effectors. The enhanced cytotoxic activity of IL-2-activated NK cells results from potent induction of the killing apparatus (13,15) and may also result from the lowering of the NK cell activation threshold by alterations in the relative expression of inhibitory and activating receptors and/ or their associated signaling components. Mono cells expressing different levels of MHC class I were generated (using b 2 m-siRNA/IFN-g) and used in killing assays with IL-2-activated NK cells.…”

Section: Resistant Tc32mentioning

confidence: 99%

A Human NK Cell Activation/Inhibition Threshold Allows Small Changes in the Target Cell Surface Phenotype To Dramatically Alter Susceptibility to NK Cells

Holmes

El-Sherbiny

Davison

et al. 2011

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

NK cell activation is negatively regulated by the expression of target cell MHC class I molecules. We show that this relationship is nonlinear due to an NK cell activation/inhibition threshold. Ewing’s sarcoma family tumor cell monolayers, which were highly susceptible to NK cells in vitro, developed a highly resistant phenotype when cultured as three-dimensional multicellular tumor spheroid structures. This suggested that tumor architecture is likely to influence the susceptibility to NK cells in vivo. Resistance of the multicellular tumor spheroid was associated with the increased expression of MHC class I molecules and greatly reduced NK cell activation, implying that a threshold of NK cell activation/inhibition had been crossed. Reducing MHC class I expression on Ewing’s sarcoma family tumor monolayers did not alter their susceptibility to NK cells, whereas increased expression of MHC class I rendered them resistant and allowed the threshold point to be identified. This threshold, as defined by MHC class I expression, was predictive of the number of NK-resistant target cells within a population. A threshold permits modest changes in the target cell surface phenotype to profoundly alter the susceptibility to NK cells. Whereas this allows for the efficient detection of target cells, it also provides a route for pathogens and tumors to evade NK cell attack.

show abstract

“…PFN is a pore-forming glycoprotein that can bind to phospholipid components of target cell membranes in the presence of Ca 2+ ions and subsequently oligomerizes to form pores with a diameter of 5-20 nm (142)(143)(144)(145). Contrary to progranzymes, which are activated by dipeptidyl peptidase I (DPPI), proteolytic processing of the perforin precursor occurs in the absence of DPPI activity (98). There are two well-founded hypotheses explaining the process of the PFNassisted GrB entry into the cytoplasm of target cells.…”

Section: Mechanism Of Granzyme B Internalization Into Target Cellsmentioning

confidence: 99%

Granzyme B-induced apoptosis in cancer cells and its regulation (Review)

Krepela¹

2010

Int J Oncol

View full text Add to dashboard Cite

Abstract. The granzyme B-induced cell death has been traditionally viewed as a primary mechanism that is used by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells to eliminate harmful target cells including allogeneic, virally infected and tumour cells. Granzyme B (GrB) is the most abundant serine protease which is stored in secretory granules of CTLs and NK cells. After recognition of the target cell, the engaged CTLs and NK cells vectorially secrete GrB along with other granule proteins including perforin into the immunological synapse. From this submicroscopic intercellular cleft GrB translocates into the cytoplasm of the target cell. Although several models have been proposed to explain the GrB delivery mechanism, conclusive understanding of this process remains still elusive. Once in the cytoplasm, GrB cleaves and activates, or inactivates, multiple protein substrates, resulting eventually into apoptotic demise of the target cell. This review is focused on the gene structure and expression of GrB, its biosynthesis and activation, delivery mechanisms into the target cell cytoplasm, direct proteolytic involvement in activation of several pro-apoptotic pathways, and on regulation of its activity in cancer cells. Moreover, emphasis is given to the GrB-mediated anticancer effects and future clinical applications of the GrB-based and tumour-targeted recombinant fusion constructs. Contents1. Introduction 2. Granzyme B gene organization and regulation of expression 3. Granzyme B biosynthesis, subcellular localization and activation 4. Granzyme B structure and substrate specificity 5. Granzyme B delivery mechanism into the target cell cytoplasm 6. Death pathways activated by granzyme B in cancer cells 7. Regulation of granzyme B activity 8. Granzyme B and anticancer therapy 9. Conclusion and the future directions of research IntroductionSusceptibility of tumour cells to apoptotic death depends on their capabilities to express the components of apoptosis pathways and to activate them in response to extrinsic or intrinsic death signals. The extrinsic death pathways are represented by the death receptor-and the cytotoxic granulemediated pathways. On the other hand, the intrinsic death mechanisms are represented by the mitochondrial, lysosomal and PIDDosome death pathways.The death receptor pathway is triggered by the binding of a death ligand, such as FasL (also known as CD95L) and TRAIL (also known as APO-2L) to a specific transmembrane death receptors, Fas (also known as APO-1/CD95) and death receptor 4 and/or 5 (DR4, DR5), respectively (1,2). After binding of the cytosolic Fas-associated death domain adaptor protein (FADD, also known as MORT1) to the liganded death receptors, the initiator procaspase-8 and/or -10 are bound to engaged FADD completing the formation of the deathinducing signalling complexes (DISCs) (1-4). Within DISCs, procaspase-8 and -10 are activated via homodimerization and the active caspase-8 and -10, arising through interdimer proteolytic processing, dissociate from DISCs into the cyt...

show abstract

Proteolytic Activation of the Cytotoxic Phenotype during Human NK Cell Development

Cited by 19 publications

References 60 publications

Blocking oncogenic RAS enhances tumour cell surface MHC class I expression but does not alter susceptibility to cytotoxic lymphocytes

Blocking oncogenic RAS enhances tumour cell surface MHC class I expression but does not alter susceptibility to cytotoxic lymphocytes

A Human NK Cell Activation/Inhibition Threshold Allows Small Changes in the Target Cell Surface Phenotype To Dramatically Alter Susceptibility to NK Cells

Granzyme B-induced apoptosis in cancer cells and its regulation (Review)

Contact Info

Product

Resources

About