The cytotoxic protease granzyme M is expressed by lymphocytes of both the innate and adaptive immune system

Koning, Pieter J.A. de; Tesselaar, Kiki; Bovenschen, Niels; Çolak, Selçuk; Quadir, Razi; Volman, Thomas; Kummer, J. Alain

doi:10.1016/j.molimm.2009.10.001

Cited by 29 publications

(27 citation statements)

References 39 publications

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“…GrM is highly expressed in NK cells (18,19), and its expression in LAK cells was confirmed by FACS analysis (data not shown). HeLa cells coexpressing recombinant pp71 and GFP were challenged with LAK cells for 4 h, after which lysates were analyzed for pp71 proteolysis (Fig.…”

Section: Grm Cleaves Pp71 In Living Cellsmentioning

confidence: 79%

“…Although GrA and GrB have been well studied, little is known about the functions of the other human Grs. GrM, which is highly expressed by NK cells (18,19), induces a specialized mode of cell death that affects mitochondrial integrity (20)(21)(22)(23)(24). GrM-deficient mice display increased susceptibility to murine CMV (MCMV) infection, indicating the physiological importance of GrM in CMV clearance in vivo (25).…”

Section: Cd8mentioning

confidence: 99%

“…Primary Abs to the following proteins were obtained from commercial sources: GFP (mouse monoclonal; Roche, Basel, Switzerland), hemagglutinin (HA, mouse monoclonal; Sigma-Aldrich, St. Louis, MO), b-tubulin (mouse monoclonal; SigmaAldrich), IE1 (mouse monoclonal, Argene), pp65 (mouse monoclonal, 1-L-11; Santa Cruz Biotechnology, Santa Cruz, CA), and pp71 (goat polyclonal, vC-20; Santa Cruz Biotechnology). Mouse mAb against pp71 (2H10-9 and 10G11) are provided by Dr. T. Shenk (Princeton University, Princeton, NJ), and mouse mAbs against GrM (3D4B7) have been described previously (19,32). Secondary HRP-conjugated goat anti-mouse and rabbit anti-goat Abs were purchased from BioSource International (Camarillo, CA) and DakoCytomation (Glostrup, Denmark), respectively.…”

Section: Virus and Absmentioning

confidence: 99%

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Noncytotoxic Inhibition of Cytomegalovirus Replication through NK Cell Protease Granzyme M-Mediated Cleavage of Viral Phosphoprotein 71

Domselaar¹,

Philippen²,

Quadir³

et al. 2010

The Journal of Immunology

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Granzyme M (GrM) is highly expressed in cytotoxic granules of NK cells, which provide the first line of defense against viral pathogens. GrM knockout mice show increased susceptibility toward murine CMV infection. Although GrM is a potent inducer of cell death, the mechanism by which GrM eliminates viruses remains elusive. In this paper, we show that purified human GrM in combination with the perforin-analog streptolysin O (SLO) strongly inhibited human CMV (HCMV) replication in fibroblasts in the absence of host cell death. In a proteomic approach, GrM was highly specific toward the HCMV proteome and most efficiently cleaved phosphoprotein 71 (pp71), an HCMV tegument protein that is critical for viral replication. Cleavage of pp71 occurred when viral lysates were incubated with purified GrM, when intact cells expressing recombinant pp71 were challenged with living cytotoxic effector cells, and when HCMV-infected fibroblasts were incubated with SLO and purified GrM. GrM directly cleaved pp71 after Leu439, which coincided with aberrant cellular localization of both pp71 cleavage fragments as determined by confocal immunofluorescence. In a luciferase reporter assay, cleavage of pp71 after Leu439 by GrM completely abolished the ability of pp71 to transactivate the HCMV major immediate-early promoter, which is indispensable for effective HCMV replication. Finally, GrM decreased immediate-early 1 protein expression in HCMV-infected fibroblasts. These results indicate that the NK cell protease GrM mediates cell death-independent antiviral activity by direct cleavage of a viral substrate.

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Section: Grm Cleaves Pp71 In Living Cellsmentioning

confidence: 79%

Section: Cd8mentioning

confidence: 99%

Section: Virus and Absmentioning

confidence: 99%

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Noncytotoxic Inhibition of Cytomegalovirus Replication through NK Cell Protease Granzyme M-Mediated Cleavage of Viral Phosphoprotein 71

Domselaar¹,

Philippen²,

Quadir³

et al. 2010

The Journal of Immunology

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“…Alexa Fluor 488-conjugated anti-GrM (clone 4B2G4) was generated as described previously. 41 Cell culture and cell-free protein lysates. Cells were cultured in a 5% CO 2 atmosphere at 371C.…”

Section: Discussionmentioning

confidence: 99%

Granzyme M targets host cell hnRNP K that is essential for human cytomegalovirus replication

et al. 2012

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Human cytomegalovirus (HCMV) is the most frequent viral cause of congenital defects and HCMV infection in immunocompromised patients may trigger devastating disease. Cytotoxic lymphocytes control HCMV by releasing granzymes towards virus-infected cells. In mice, granzyme M (GrM) has a physiological role in controlling murine CMV infection. However, the underlying mechanism remains poorly understood. In this study, we showed that human GrM was expressed by HCMV-specific CD8 þ T cells both in latently infected healthy individuals and in transplant patients during primary HCMV infection. We identified host cell heterogeneous nuclear ribonucleoprotein K (hnRNP K) as a physiological GrM substrate. GrM most efficiently cleaved hnRNP K in the presence of RNA at multiple sites, thereby likely destroying hnRNP K function. Host cell hnRNP K was essential for HCMV replication not only by promoting viability of HCMV-infected cells but predominantly by regulating viral immediate-early 2 (IE2) protein levels. Furthermore, hnRNP K interacted with IE2 mRNA. Finally, GrM decreased IE2 protein expression in HCMV-infected cells. Our data suggest that targeting of hnRNP K by GrM contributes to the mechanism by which cytotoxic lymphocytes inhibit HCMV replication. This is the first evidence that cytotoxic lymphocytes target host cell proteins to control HCMV infections.

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“…cd-T cell-mediated cytotoxicity is carried out through dozens of pathways, including Fas-Fas ligand interaction and secretion of perforin, granzyme B 27 and granzyme M. 94 In contrast to the cytotoxicity of NK cells, the cytotoxicity mediated by Vc9Vd2 cd-T cells 22 and Vd2 2 cd-T cells 34 against influenza virus-and HCMV-infected cells can be initiated independent of the antibody-dependent cellmediated cytotoxicity effect. Consistent with this mechanism, the Fc receptor CD16, a key player in antibody-dependent cell-mediated cytotoxicity, has been shown to improve the anti-viral activity of Vd2 2 cd-T cells by inducing them to produce IFN-c during HCMV infection.…”

Section: Direct Anti-infection Activity Mediated By Cd-t Cells Directmentioning

confidence: 99%

γδ-T cells: an unpolished sword in human anti-infection immunity

et al. 2012

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cd-T cells represent a small population of immune cells, but play an indispensable role in host defenses against exogenous pathogens, immune surveillance of endogenous pathogenesis and even homeostasis of the immune system. Activation and expansion of cd-T cells are generally observed in diverse human infectious diseases and correlate with their progression and prognosis. cd-T cells have both 'innate' and 'adaptive' characteristics in the immune response, and their anti-infection activities are mediated by multiple pathways that are under elaborate regulation by other immune components. In this review, we summarize the current state of the literature and the recent advancements in cd-T cell-mediated immune responses against common human infectious pathogens. Although further investigation is needed to improve our understanding of the characteristics of different cd-T cell subpopulations under specific conditions, cd-T cell-based therapy has great potential for the treatment of infectious diseases.

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The cytotoxic protease granzyme M is expressed by lymphocytes of both the innate and adaptive immune system

Cited by 29 publications

References 39 publications

Noncytotoxic Inhibition of Cytomegalovirus Replication through NK Cell Protease Granzyme M-Mediated Cleavage of Viral Phosphoprotein 71

Noncytotoxic Inhibition of Cytomegalovirus Replication through NK Cell Protease Granzyme M-Mediated Cleavage of Viral Phosphoprotein 71

Granzyme M targets host cell hnRNP K that is essential for human cytomegalovirus replication

γδ-T cells: an unpolished sword in human anti-infection immunity

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