Abstract
Granzymes (granule-secreted enzymes) are a family of serine proteases that have been viewed as redundant cytotoxic enzymes since their discovery more than 30 years ago. Predominantly produced by cytotoxic lymphocytes and natural killer cells, granzymes are delivered into the cytoplasm of target cells through immunological synapses in cooperation with the pore-forming protein perforin. After internalization, granzymes can initiate cell death through the cleavage of intracellular substrates. However, evidence now also demonstrates the existence of non-cytotoxic, pro-inflammatory, intracellular and extracellular functions that are granzyme specific. Under pathological conditions, granzymes can be produced and secreted extracellularly by immune cells as well as by non-immune cells. Depending on the granzyme, accumulation in the extracellular milieu might contribute to inflammation, tissue injury, impaired wound healing, barrier dysfunction, osteoclastogenesis and/or autoantigen generation.
Key points
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Granzymes are serine proteases with both cytotoxic and non-cytotoxic functions; phenotypic and mechanistic characterization of granzymes in rheumatic diseases is needed to delineate their specific roles.
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The five human granzymes have unique substrate specificities and functional roles, as determined by granzyme-specific cleavage preferences, location of accumulation (intracellular or extracellular) and exposure to substrates in tissues.
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Extracellular granzyme activity can contribute to tissue injury, inflammation, autoimmunity, epithelial and endothelial barrier dysfunction, bullae formation, impaired wound healing, and degenerative or pathological aging.
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In addition to cytoplasmic proteins involved in apoptosis, granzyme B substrates include extracellular matrix proteins, hemidesmosomal or desmosomal proteins, pro-inflammatory cytokines, cell surface receptors and autoantigens.
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Granzyme A and granzyme B are elevated in synovial fluid, tissues and plasma of people with rheumatoid arthritis; in an arthritis model, Gzma−/− mice have lower disease severity than that of wild-type mice.
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CD8+ T cells expressing granzyme B and granzyme K are enriched in the peripheral blood and inflamed tissues of people with rheumatoid arthritis, systemic lupus erythematosus or Sjögren syndrome.
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References
Joeckel, L. T. & Bird, P. I. Are all granzymes cytotoxic in vivo? Biol. Chem. 395, 181–202 (2014).
Voskoboinik, I., Whisstock, J. C. & Trapani, J. A. Perforin and granzymes: function, dysfunction and human pathology. Nat. Rev. Immunol. 15, 388–400 (2015).
Cullen, S. P., Adrain, C., Lüthi, A. U., Duriez, P. J. & Martin, S. J. Human and murine granzyme B exhibit divergent substrate preferences. J. Cell Biol. 176, 435–444 (2007).
Kaiserman, D. et al. The major human and mouse granzymes are structurally and functionally divergent. J. Cell Biol. 175, 619–630 (2006).
Pardo, J. et al. Granzyme B-induced cell death exerted by ex vivo CTL: discriminating requirements for cell death and some of its signs. Cell Death Differ. 15, 567–579 (2008).
Thomas, D. A., Scorrano, L., Putcha, G. V., Korsmeyer, S. J. & Ley, T. J. Granzyme B can cause mitochondrial depolarization and cell death in the absence of BID, BAX, and BAK. Proc. Natl Acad. Sci. USA 98, 14985–14990 (2001).
Sánchez-Martínez, D. et al. Human NK cells activated by EBV+ lymphoblastoid cells overcome anti-apoptotic mechanisms of drug resistance in haematological cancer cells. Oncoimmunology 4, e991613 (2015).
Plasman, K., Demol, H., Bird, P. I., Gevaert, K. & Van Damme, P. Substrate specificities of the granzyme tryptases A and K. J. Proteome Res. 13, 6067–6077 (2014).
Plasman, K. et al. Conservation of the extended substrate specificity profiles among homologous granzymes across species. Mol. Cell Proteom. 12, 2921–2934 (2013).
Garzón-Tituaña, M. et al. Granzyme A inhibition reduces inflammation and increases survival during abdominal sepsis. Theranostics 11, 3781–3795 (2021).
Metkar, S. S. et al. Human and mouse granzyme A induce a proinflammatory cytokine response. Immunity 29, 720–733 (2008).
Hagn, M. et al. Human B cells secrete granzyme B when recognizing viral antigens in the context of the acute phase cytokine IL-21. J. Immunol. 183, 1838–1845 (2009).
Hink-Schauer, C., Estébanez-Perpiñá, E., Kurschus, F. C., Bode, W. & Jenne, D. E. Crystal structure of the apoptosis-inducing human granzyme A dimer. Nat. Struct. Biol. 10, 535–540 (2003).
Bell, J. K. et al. The oligomeric structure of human granzyme A is a determinant of its extended substrate specificity. Nat. Struct. Biol. 10, 527–534 (2003).
Lieberman, J. Granzyme A activates another way to die. Immunol. Rev. 235, 93–104 (2010).
Pardo, J. et al. The biology of cytotoxic cell granule exocytosis pathway: granzymes have evolved to induce cell death and inflammation. Microbes Infect. 11, 452–459 (2009).
Beresford, P. J. et al. Granzyme A activates an endoplasmic reticulum-associated caspase-independent nuclease to induce single-stranded DNA nicks. J. Biol. Chem. 276, 43285–43293 (2001).
Zhou, Z. et al. Granzyme A from cytotoxic lymphocytes cleaves GSDMB to trigger pyroptosis in target cells. Science 368, eaaz7548 (2020).
Liu, X., Xia, S., Zhang, Z., Wu, H. & Lieberman, J. Channelling inflammation: gasdermins in physiology and disease. Nat. Rev. Drug Discov. 20, 384–405 (2021).
Isaaz, S., Baetz, K., Olsen, K., Podack, E. & Griffiths, G. M. Serial killing by cytotoxic T lymphocytes: T cell receptor triggers degranulation, re-filling of the lytic granules and secretion of lytic proteins via a non-granule pathway. Eur. J. Immunol. 25, 1071–1079 (1995).
Nakamura, K., Arahata, K., Ishiura, S., Osame, M. & Sugita, H. Degradative activity of granzyme A on skeletal muscle proteins in vitro: a possible molecular mechanism for muscle fiber damage in polymyositis. Neuromuscul. Dis. 3, 303–310 (1993).
Campbell, R. A. et al. Granzyme A in human platelets regulates the synthesis of proinflammatory cytokines by monocytes in aging. J. Immunol. 200, 295–304 (2018).
Arias, M. A. et al. Elucidating sources and roles of granzymes A and B during bacterial infection and sepsis. Cell Rep. 8, 420–429 (2014).
Santiago, L. et al. Extracellular granzyme A promotes colorectal cancer development by enhancing gut inflammation. Cell Rep. 32, 107847 (2020).
Santiago, L. et al. Granzyme A contributes to inflammatory arthritis in mice through stimulation of osteoclastogenesis. Arthritis Rheumatol. 69, 320–334 (2017).
Masson, D. & Tschopp, J. Inhibition of lymphocyte protease granzyme A by antithrombin III. Mol. Immunol. 25, 1283–1289 (1988).
Simon, M. M., Tran, T., Fruth, U., Gurwitz, D. & Kramer, M. D. Regulation of mouse T cell associated serine proteinase-1 (MTSP-1) by proteinase inhibitors and sulfated polysaccharides. Biol. Chem. Hoppe Seyler 371, 81–87 (1990).
Kaiserman, D. et al. Identification of serpinb6b as a species-specific mouse granzyme A inhibitor suggests functional divergence between human and mouse granzyme A. J. Biol. Chem. 289, 9408–9417 (2014).
Aybay, E. et al. Extended cleavage specificities of human granzymes A and K, two closely related enzymes with conserved but still poorly defined functions in T and NK cell-mediated immunity. Front. Immunol. 14, 1211295 (2023).
Boivin, W. A., Cooper, D. M., Hiebert, P. R. & Granville, D. J. Intracellular versus extracellular granzyme B in immunity and disease: challenging the dogma. Lab. Invest. 89, 1195–1220 (2009).
Thornberry, N. A. et al. A combinatorial approach defines specificities of members of the caspase family and granzyme B: functional relationships established for key mediators of apoptosis. J. Biol. Chem. 272, 17907–17911 (1997).
Zhang, Z. et al. Gasdermin E suppresses tumour growth by activating anti-tumour immunity. Nature 579, 415–420 (2020).
Aubert, A., Lane, M., Jung, K. & Granville, D. J. Granzyme B as a therapeutic target: an update in 2022. Expert Opin. Ther. Targets 26, 979–993 (2022).
Richardson, K. C., Jung, K., Pardo, J., Turner, C. T. & Granville, D. J. Noncytotoxic roles of granzymes in health and disease. Physiology 37, 323–348 (2022).
Prakash, M. D., Bird, C. H. & Bird, P. I. Active and zymogen forms of granzyme B are constitutively released from cytotoxic lymphocytes in the absence of target cell engagement. Immunol. Cell Biol. 87, 249–254 (2009).
Jung, K., Pawluk, M. A., Lane, M., Nabai, L. & Granville, D. J. Granzyme B in epithelial barrier dysfunction and related skin diseases. Am. J. Physiol. Cell Physiol. 323, C170–c189 (2022).
Skjelland, M. et al. Plasma levels of granzyme B are increased in patients with lipid-rich carotid plaques as determined by echogenicity. Atherosclerosis 195, e142–e146 (2007).
Tak, P. P. et al. The levels of soluble granzyme A and B are elevated in plasma and synovial fluid of patients with rheumatoid arthritis (RA). Clin. Exp. Immunol. 116, 366–370 (1999).
Hodge, S., Hodge, G., Nairn, J., Holmes, M. & Reynolds, P. N. Increased airway granzyme B and perforin in current and ex-smoking COPD subjects. COPD 3, 179–187 (2006).
Hiroyasu, S. et al. Granzyme B inhibition reduces disease severity in autoimmune blistering diseases. Nat. Commun. 12, 302 (2021).
Malmestrom, C. et al. Relapses in multiple sclerosis are associated with increased CD8+ T-cell mediated cytotoxicity in CSF. J. Neuroimmunol. 196, 159–165 (2008).
Turner, C. T. et al. Granzyme B contributes to barrier dysfunction in oxazolone-induced skin inflammation through E-cadherin and FLG cleavage. J. Invest. Dermatol. 141, 36–47 (2021).
Turner, C. T. et al. Granzyme B mediates impaired healing of pressure injuries in aged skin. NPJ Aging Mech. Dis. 7, 6 (2021).
Russo, V. et al. Granzyme B is elevated in autoimmune blistering diseases and cleaves key anchoring proteins of the dermal-epidermal junction. Sci. Rep. 8, 9690 (2018).
Kurschus, F. C. et al. Killing of target cells by redirected granzyme B in the absence of perforin. FEBS Lett. 562, 87–92 (2004).
Tremblay, G. M., Wolbink, A. M., Cormier, Y. & Hack, C. E. Granzyme activity in the inflamed lung is not controlled by endogenous serine proteinase inhibitors. J. Immunol. 165, 3966–3969 (2000).
Buzza, M. S. et al. Extracellular matrix remodeling by human granzyme B via cleavage of vitronectin, fibronectin, and laminin. J. Biol. Chem. 280, 23549–23558 (2005).
Prakash, M. D. et al. Granzyme B promotes cytotoxic lymphocyte transmigration via basement membrane remodeling. Immunity 41, 960–972 (2014).
Chamberlain, C. M. et al. Perforin-independent extracellular granzyme B activity contributes to abdominal aortic aneurysm. Am. J. Pathol. 176, 1038–1049 (2010).
Ang, L. S. et al. Serpina3n attenuates granzyme B-mediated decorin cleavage and rupture in a murine model of aortic aneurysm. Cell Death Dis. 2, e209 (2011).
Hiebert, P. R. et al. Granzyme B contributes to extracellular matrix remodeling and skin aging in apolipoprotein E knockout mice. Exp. Gerontol. 46, 489–499 (2011).
Boivin, W. A. et al. Granzyme B cleaves decorin, biglycan and soluble betaglycan, releasing active transforming growth factor-β1. PLoS One 7, e33163 (2012).
Hendel, A. & Granville, D. J. Granzyme B cleavage of fibronectin disrupts endothelial cell adhesion, migration and capillary tube formation. Matrix Biol. 32, 14–22 (2013).
Hiebert, P. R., Wu, D. & Granville, D. J. Granzyme B degrades extracellular matrix and contributes to delayed wound closure in apolipoprotein E knockout mice. Cell Death Differ. 20, 1404–1414 (2013).
Hendel, A., Hsu, I. & Granville, D. J. Granzyme B releases vascular endothelial growth factor from extracellular matrix and induces vascular permeability. Lab. Invest. 94, 716–725 (2014).
Parkinson, L. G. et al. Granzyme B mediates both direct and indirect cleavage of extracellular matrix in skin after chronic low-dose ultraviolet light irradiation. Aging Cell 14, 67–77 (2015).
Shen, Y. et al. Topical small molecule granzyme B inhibitor improves remodeling in a murine model of impaired burn wound healing. Exp. Mol. Med. 50, 1–11 (2018).
Matsubara, J. A. et al. Retinal distribution and extracellular activity of granzyme B: a serine protease that degrades retinal pigment epithelial tight junctions and extracellular matrix proteins. Front. Immunol. 11, 574 (2020).
Obasanmi, G. et al. Granzyme B contributes to choroidal neovascularization and age-related macular degeneration through proteolysis of thrombospondin-1. Lab. Invest. 103, 100123 (2023).
Ronday, H. K. et al. Human granzyme B mediates cartilage proteoglycan degradation and is expressed at the invasive front of the synovium in rheumatoid arthritis. Rheumatology 40, 55–61 (2001).
Froelich, C. J. et al. Human granzyme B degrades aggrecan proteoglycan in matrix synthesized by chondrocytes. J. Immunol. 151, 7161–7171 (1993).
Buzza, M. S. et al. Antihemostatic activity of human granzyme B mediated by cleavage of von Willebrand factor. J. Biol. Chem. 283, 22498–22504 (2008).
Qian, Q. et al. Maternal diesel particle exposure promotes offspring asthma through NK cell-derived granzyme B. J. Clin. Invest. 130, 4133–4151 (2020).
Loeb, C. R. K., Harris, J. L. & Craik, C. S. Granzyme B proteolyzes receptors important to proliferation and survival, tipping the balance toward apoptosis. J. Biol. Chem. 281, 28326–28335 (2006).
Shen, Y. et al. Granzyme B deficiency protects against angiotensin II-induced cardiac fibrosis. Am. J. Pathol. 186, 87–100 (2016).
Afonina, I. S. et al. Granzyme B-dependent proteolysis acts as a switch to enhance the proinflammatory activity of IL-1α. Mol. Cell 44, 265–278 (2011).
Omoto, Y. et al. Granzyme B is a novel interleukin-18 converting enzyme. J. Dermatol. Sci. 59, 129–135 (2010).
Lim, Y. S. et al. NK cell-derived extracellular granzyme B drives epithelial ulceration during HSV-2 genital infection. Cell Rep. 42, 112410 (2023).
Hu, M. D. et al. γδ intraepithelial lymphocytes facilitate pathological epithelial cell shedding via CD103-mediated granzyme release. Gastroenterology 162, 877–889.e877 (2022).
Casciola-Rosen, L., Andrade, F., Ulanet, D., Wong, W. B. & Rosen, A. Cleavage by granzyme B is strongly predictive of autoantigen status: implications for initiation of autoimmunity. J. Exp. Med. 190, 815–826 (1999).
Darrah, E. & Rosen, A. Granzyme B cleavage of autoantigens in autoimmunity. Cell Death Differ. 17, 624–632 (2010).
Darrah, E. et al. Proteolysis by granzyme B enhances presentation of autoantigenic peptidylarginine deiminase 4 epitopes in rheumatoid arthritis. J. Proteome Res. 16, 355–365 (2017).
Haddad, P. et al. Structure and evolutionary origin of the human granzyme H gene. Int. Immunol. 3, 57–66 (1991).
Wang, L. et al. Structural insights into the substrate specificity of human granzyme H: the functional roles of a novel RKR motif. J. Immunol. 188, 765–773 (2012).
Sedelies, K. A. et al. Discordant regulation of granzyme H and granzyme B expression in human lymphocytes. J. Biol. Chem. 279, 26581–26587 (2004).
Rönnberg, E. et al. Granzyme H is a novel protease expressed by human mast cells. Int. Arch. Allergy Immunol. 165, 68–74 (2014).
Waterhouse, N. J. & Trapani, J. A. H is for helper: granzyme H helps granzyme B kill adenovirus-infected cells. Trends Immunol. 28, 373–375 (2007).
Romero, V., Fellows, E., Jenne, D. E. & Andrade, F. Cleavage of La protein by granzyme H induces cytoplasmic translocation and interferes with La-mediated HCV-IRES translational activity. Cell Death Differ. 16, 340–348 (2009).
Yu, D. et al. Natural killer cells disrupt nerve fibers by granzyme H in atheriosclerotic cerebral small vessel disease. J. Gerontol. A Biol. Sci. Med. Sci. 78, 414–423 (2023).
Bovenschen, N. et al. Granzyme K displays highly restricted substrate specificity that only partially overlaps with granzyme A. J. Biol. Chem. 284, 3504–3512 (2009).
Hink-Schauer, C. et al. The 2.2-A crystal structure of human pro-granzyme K reveals a rigid zymogen with unusual features. J. Biol. Chem. 277, 50923–50933 (2002).
Bratke, K., Kuepper, M., Bade, B., Virchow, J. C. Jr & Luttmann, W. Differential expression of human granzymes A, B, and K in natural killer cells and during CD8+ T cell differentiation in peripheral blood. Eur. J. Immunol. 35, 2608–2616 (2005).
Duquette, D. et al. Human granzyme K Is a feature of innate T cells in blood, tissues, and tumors, responding to cytokines rather than TCR stimulation. J. Immunol. 211, 633–647 (2023).
Bade, B. et al. Detection of soluble human granzyme K in vitro and in vivo. Eur. J. Immunol. 35, 2940–2948 (2005).
Rucevic, M. et al. Altered levels and molecular forms of granzyme K in plasma from septic patients. Shock 27, 488–493 (2007).
Li, T., Yang, C., Jing, J., Sun, L. & Yuan, Y. Granzyme K — a novel marker to identify the presence and rupture of abdominal aortic aneurysm. Int. J. Cardiol. 338, 242–247 (2021).
Bratke, K. et al. Granzyme K: a novel mediator in acute airway inflammation. Thorax 63, 1006–1011 (2008).
Joeckel, L. T. et al. Mouse granzyme K has pro-inflammatory potential. Cell Death Differ. 18, 1112–1119 (2011).
Turner, C. T. et al. Granzyme K expressed by classically activated macrophages contributes to inflammation and impaired remodeling. J. Invest. Dermatol. 139, 930–939 (2019).
Sharma, M. et al. Extracellular granzyme K mediates endothelial activation through the cleavage of protease-activated receptor-1. FEBS J. 283, 1734–1747 (2016).
Kaiserman, D. et al. Granzyme K initiates IL-6 and IL-8 release from epithelial cells by activating protease-activated receptor 2. PLoS One 17, e0270584 (2022).
Jonsson, A. H. et al. Granzyme K+ CD8 T cells form a core population in inflamed human tissue. Sci. Transl. Med. 14, eabo0686 (2022).
Turner, C. T. et al. Granzyme K contributes to endothelial microvascular damage and leakage during skin inflammation. Br. J. Dermatol. 189, 279–291 (2023).
Mogilenko, D. A. et al. Comprehensive profiling of an aging immune system reveals clonal GZMK+ CD8+ T cells as conserved hallmark of inflammaging. Immunity 54, 99–115.e112 (2021).
Verschoor, C. P. et al. NK- and T-cell granzyme B and K expression correlates with age, CMV infection and influenza vaccine-induced antibody titres in older adults. Front. Aging 3, 1098200 (2022).
Tyrrell, D. J. et al. Clonally expanded memory CD8+ T cells accumulate in atherosclerotic plaques and are pro-atherogenic in aged mice. Nat. Aging 3, 1576–1590 (2023).
Fernandez, D. M. et al. Single-cell immune landscape of human atherosclerotic plaques. Nat. Med. 25, 1576–1588 (2019).
Cai, Y. et al. Single-cell immune profiling reveals functional diversity of T cells in tuberculous pleural effusion. J. Exp. Med. 219, e20211777 (2022).
Xu, T. et al. Single-cell profiling reveals pathogenic role and differentiation trajectory of granzyme K+CD8+ T cells in primary Sjögren’s syndrome. JCI Insight 8, e167490 (2023).
Arazi, A. et al. The immune cell landscape in kidneys of patients with lupus nephritis. Nat. Immunol. 20, 902–914 (2019).
Dunlap, G. S. et al. Single-cell transcriptomics reveals distinct effector profiles of infiltrating T cells in lupus skin and kidney. JCI Insight 7, e156341 (2022).
de Poot, S. A. H. et al. Human and mouse granzyme M display divergent and species-specific substrate specificities. Biochem. J. 437, 431–442 (2011).
Sayers, T. J. et al. The restricted expression of granzyme M in human lymphocytes. J. Immunol. 166, 765–771 (2001).
de Poot, S. A. & Bovenschen, N. Granzyme M: behind enemy lines. Cell Death Differ. 21, 359–368 (2014).
Pao, L. I. et al. Functional analysis of granzyme M and its role in immunity to infection. J. Immunol. 175, 3235–3243 (2005).
Anthony, D. A. et al. A role for granzyme M in TLR4-driven inflammation and endotoxicosis. J. Immunol. 185, 1794–1803 (2010).
Baschuk, N. et al. NK cell intrinsic regulation of MIP-1ɑ by granzyme M. Cell Death Dis. 5, e1115 (2014).
Nordstrom, D. C. et al. Granzyme A-immunoreactive cells in synovial fluid in reactive and rheumatoid arthritis. Clin. Rheumatol. 11, 529–532 (1992).
Smeets, T. J., Dolhain, R. J., Breedveld, F. C. & Tak, P. P. Analysis of the cellular infiltrates and expression of cytokines in synovial tissue from patients with rheumatoid arthritis and reactive arthritis. J. Pathol. 186, 75–81 (1998).
Qiao, J. et al. Elevated serum granzyme B levels are associated with disease activity and joint damage in patients with rheumatoid arthritis. J. Int. Med. Res. 48, 0300060520962954 (2020).
Goldbach-Mansky, R. et al. Raised granzyme B levels are associated with erosions in patients with early rheumatoid factor positive rheumatoid arthritis. Ann. Rheum. Dis. 64, 715–721 (2005).
Knevel, R. et al. A genetic variant in granzyme B is associated with progression of joint destruction in rheumatoid arthritis. Arthritis Rheuma. 65, 582–589 (2013).
Colombo, E., Scarsi, M., Piantoni, S., Tincani, A. & Airo, P. Serum levels of granzyme B decrease in patients with rheumatoid arthritis responding to abatacept. Clin. Exp. Rheumatol. 34, 37–41 (2016).
Zhang, F. et al. Defining inflammatory cell states in rheumatoid arthritis joint synovial tissues by integrating single-cell transcriptomics and mass cytometry. Nat. Immunol. 20, 928–942 (2019).
Moon, J.-S. et al. Cytotoxic CD8+ T cells target citrullinated antigens in rheumatoid arthritis. Nat. Commun. 14, 319 (2023).
Tak, P. P. et al. Granzyme-positive cytotoxic cells are specifically increased in early rheumatoid synovial tissue. Arthritis Rheum. 37, 1735–1743 (1994).
Kim, W. J., Kim, H., Suk, K. & Lee, W. H. Macrophages express granzyme B in the lesion areas of atherosclerosis and rheumatoid arthritis. Immunol. Lett. 111, 57–65 (2007).
Xu, L. et al. Impairment of granzyme B-producing regulatory B cells correlates with exacerbated rheumatoid arthritis. Front. Immunol. 8, 768 (2017).
Cui, D. et al. Changes in regulatory B cells and their relationship with rheumatoid arthritis disease activity. Clin. Exp. Med. 15, 285–292 (2015).
Kageyama, Y., Kobayashi, H., Kato, N. & Shimazu, M. Etanercept reduces the serum levels of macrophage chemotactic protein-1 in patients with rheumatoid arthritis. Mod. Rheumatol. 19, 372–378 (2009).
Horiuchi, K., Saito, S., Sasaki, R., Tomatsu, T. & Toyama, Y. Expression of granzyme B in human articular chondrocytes. J. Rheumatol. 30, 1799–1810 (2003).
Darrah, E. et al. Erosive rheumatoid arthritis is associated with antibodies that activate PAD4 by increasing calcium sensitivity. Sci. Transl. Med. 5, 186ra165 (2013).
Halvorsen, E. H. et al. Serum IgG antibodies to peptidylarginine deiminase 4 in rheumatoid arthritis and associations with disease severity. Ann. Rheum. Dis. 67, 414–417 (2008).
Kinloch, A. et al. Synovial fluid is a site of citrullination of autoantigens in inflammatory arthritis. Arthritis Rheum. 58, 2287–2295 (2008).
Luo, Y. et al. The minor collagens in articular cartilage. Protein Cell 8, 560–572 (2017).
Weng, S. S. H. et al. Sensitive determination of proteolytic proteoforms in limited microscale proteome samples. Mol. Cell Proteom. 18, 2335–2347 (2019).
Zhou, J., Tang, X., Ding, Y., An, Y. & Zhao, X. Natural killer cell activity and frequency of killer cell immunoglobulin-like receptors in children with different forms of juvenile idiopathic arthritis. Pediatr. Allergy Immunol. 24, 691–696 (2013).
Wang, M. et al. Single-cell analysis in blood reveals distinct immune cell profiles in gouty arthritis. J. Immunol. 210, 745–752 (2023).
Canete, J. D. et al. Distinct synovial immunopathology in Behcet disease and psoriatic arthritis. Arthritis Res. Ther. 11, R17 (2009).
Shan, L. et al. Increased intra-articular granzyme M may trigger local IFN-λ1/IL-29 response in rheumatoid arthritis. Clin. Exp. Rheumatol. 38, 220–226 (2020).
McLoughlin, R. M. et al. IL-6 trans-signaling via STAT3 directs T cell infiltration in acute inflammation. Proc. Natl Acad. Sci. USA 102, 9589–9594 (2005).
Young, L. H. et al. Expression of cytolytic mediators by synovial fluid lymphocytes in rheumatoid arthritis. Am. J. Pathol. 140, 1261–1268 (1992).
Kummer, J. A. et al. Expression of granzymes A and B in synovial tissue from patients with rheumatoid arthritis and osteoarthritis. Clin. Immunol. Immunopathol. 73, 88–95 (1994).
Nanki, T. et al. Migration of CX3CR1-positive T cells producing type 1 cytokines and cytotoxic molecules into the synovium of patients with rheumatoid arthritis. Arthritis Rheum. 46, 2878–2883 (2002).
Jaime, P. et al CD56+/CD16− natural killer cells expressing the inflammatory protease granzyme A are enriched in synovial fluid from patients with osteoarthritis. Osteoarthritis Cartilage 25, 1708–1718 (2017).
Wilson, J. A. et al. RNA-Seq analysis of chikungunya virus infection and identification of granzyme A as a major promoter of arthritic inflammation. PLoS Pathog. 13, e1006155 (2017).
Jia, T. et al. CRISPR/Cas13d targeting GZMA in PARs pathway regulates the function of osteoclasts in chronic apical periodontitis. Cell Mol. Biol. Lett. 28, 70 (2023).
Simon, M. M., Kramer, M. D., Prester, M. & Gay, S. Mouse T-cell associated serine proteinase 1 degrades collagen type IV: a structural basis for the migration of lymphocytes through vascular basement membranes. Immunology 73, 117–119 (1991).
Simon, M. M., Prester, M., Nerz, G., Kramer, M. D. & Fruth, U. Release of biologically active fragments from human plasma-fibronectin by murine T cell-specific proteinase 1 (TSP-1). Biol. Chem. Hoppe Seyler 369, 107–112 (1988).
Crow, M. K. Pathogenesis of systemic lupus erythematosus: risks, mechanisms and therapeutic targets. Ann. Rheum. Dis. 82, 999–1014 (2023).
Shah, D., Kiran, R., Wanchu, A. & Bhatnagar, A. Soluble granzyme B and cytotoxic T lymphocyte activity in the pathogenesis of systemic lupus erythematosus. Cell Immunol. 269, 16–21 (2011).
Kok, H. M. et al. Systemic and local granzyme B levels are associated with disease activity, kidney damage and interferon signature in systemic lupus erythematosus. Rheumatology 56, 2129–2134 (2017).
Chen, L. et al. IKZF1 polymorphisms are associated with susceptibility, cytokine levels, and clinical features in systemic lupus erythematosus. Medicine 99, e22607 (2020).
Daca, A. et al. Two systemic lupus erythematosus (SLE) global disease activity indexes — the SLE disease activity index and the systemic lupus activity measure —demonstrate different correlations with activation of peripheral blood CD4+ T cells. Hum. Immunol. 72, 1160–1167 (2011).
Blanco, P. et al. Increase in activated CD8+ T lymphocytes expressing perforin and granzyme B correlates with disease activity in patients with systemic lupus erythematosus. Arthritis Rheum. 52, 201–211 (2005).
Patel, J. et al. Multidimensional immune profiling of cutaneous lupus erythematosus in vivo stratified by patient response to antimalarials. Arthritis Rheumatol. 74, 1687–1698 (2022).
Abdou, A. G., Shoeib, M., Bakry, O. A. & El-Bality, H. Immunohistochemical expression of granzyme B and perforin in discoid lupus erythematosus. Ultrastruct. Pathol. 37, 408–416 (2013).
Fogagnolo, L. et al. Cytotoxic granules in distinct subsets of cutaneous lupus erythematosus. Clin. Exp. Dermatol. 39, 835–839 (2014).
Wenzel, J. et al. CXCR3-mediated recruitment of cytotoxic lymphocytes in lupus erythematosus profundus. J. Am. Acad. Dermatol. 56, 648–650 (2007).
Wenzel, J. et al. Scarring skin lesions of discoid lupus erythematosus are characterized by high numbers of skin-homing cytotoxic lymphocytes associated with strong expression of the type I interferon-induced protein MxA. Br. J. Dermatol. 153, 1011–1015 (2005).
Grassi, M., Capello, F., Bertolino, L., Seia, Z. & Pippione, M. Identification of granzyme B-expressing CD-8-positive T cells in lymphocytic inflammatory infiltrate in cutaneous lupus erythematosus and in dermatomyositis. Clin. Exp. Dermatol. 34, 910–914 (2009).
Lovato, B. H. et al. IL-1β and IL-17 in cutaneous lupus erythematous skin biopsies: could immunohistochemicals indicate a tendency towards systemic involvement? Bras. Dermatol. 99, 66–71 (2023).
Zhou, M. et al. JAK/STAT signaling controls the fate of CD8+CD103+ tissue-resident memory T cell in lupus nephritis. J. Autoimmun. 109, 102424 (2020).
Litvinova, E. et al. MAIT cells altered phenotype and cytotoxicity in lupus patients are linked to renal disease severity and outcome. Front. Immunol. 14, 1205405 (2023).
Murayama, G. et al. A critical role for mucosal-associated invariant T cells as regulators and therapeutic targets in systemic lupus erythematosus. Front. Immunol. 10, 2681 (2019).
Perez, R. K. et al. Single-cell RNA-seq reveals cell type-specific molecular and genetic associations to lupus. Science 376, eabf1970 (2022).
Hernandez-Pigeon, H. et al. UVA induces granzyme B in human keratinocytes through MIF: implication in extracellular matrix remodeling. J. Biol. Chem. 282, 8157–8164 (2007).
van Leeuwen, E. M. et al. Emergence of a CD4+CD28− granzyme B+, cytomegalovirus-specific T cell subset after recovery of primary cytomegalovirus infection. J. Immunol. 173, 1834–1841 (2004).
Aguilera, J. et al. Granzymes, IL-16, and poly(ADP-ribose) polymerase 1 increase during wildfire smoke exposure. J. Allergy Clin. Immunol. Glob. 2, 100093 (2023).
Tsokos, G. C., Lo, M. S., Costa Reis, P. & Sullivan, K. E. New insights into the immunopathogenesis of systemic lupus erythematosus. Nat. Rev. Rheumatol. 12, 716–730 (2016).
Lu, C. et al. Type I interferon suppresses tumor growth through activating the STAT3-granzyme B pathway in tumor-infiltrating cytotoxic T lymphocytes. J. Immunother. Cancer 7, 157 (2019).
Newby, B. N. et al. Type 1 interferons potentiate human CD8+ T-cell cytotoxicity through a STAT4- and granzyme B-dependent pathway. Diabetes 66, 3061–3071 (2017).
Turner, C. T., Lim, D. & Granville, D. J. Granzyme B in skin inflammation and disease. Matrix Biol. 75–76, 126–140 (2019).
Graham, K. L. et al. Granzyme B is dispensable for immunologic tolerance to self in a murine model of systemic lupus erythematosus. Arthritis Rheum. 52, 1684–1693 (2005).
Rabani, M. et al. IL-21 dependent granzyme B production of B-cells is decreased in patients with lupus nephritis. Clin. Immunol. 188, 45–51 (2018).
Bai, M. et al. Impaired granzyme B-producing regulatory B cells in systemic lupus erythematosus. Mol. Immunol. 140, 217–224 (2021).
Brito-Zeron, P. et al. Sjögren syndrome. Nat. Rev. Dis. Prim. 2, 16047 (2016).
Kaneko, N. et al. Cytotoxic CD8+ T cells may be drivers of tissue destruction in Sjögren’s syndrome. Sci. Rep. 12, 15427 (2022).
Akiyama, M., Yoshimoto, K. & Kaneko, Y. Significant association of CX3CR1+CD8 T cells with aging and distinct clinical features in Sjögren’s syndrome and IgG4-related disease. Clin. Exp. Rheumatol. 41, 2409–2417 (2023).
Tasaki, S. et al. Multiomic disease signatures converge to cytotoxic CD8 T cells in primary Sjögren’s syndrome. Ann. Rheum. Dis. 76, 1458–1466 (2017).
Wang, Q. et al. Correlation of peripheral CD4+GranzB+CTLs with disease severity in patients with primary Sjögren’s syndrome. Arthritis Res. Ther. 23, 257 (2021).
Huang, M. et al. Detection of apoptosis-specific autoantibodies directed against granzyme B-induced cleavage fragments of the SS-B (La) autoantigen in sera from patients with primary Sjögren’s syndrome. Clin. Exp. Immunol. 142, 148–154 (2005).
Pope, J. E. et al. State-of-the-art evidence in the treatment of systemic sclerosis. Nat. Rev. Rheumatol. 19, 212–226 (2023).
Fuschiotti, P., Medsger, T. A. Jr & Morel, P. A. Effector CD8+ T cells in systemic sclerosis patients produce abnormally high levels of interleukin-13 associated with increased skin fibrosis. Arthritis Rheum. 60, 1119–1128 (2009).
Ayano, M. et al. Increased CD226 expression on CD8+ T cells is associated with upregulated cytokine production and endothelial cell injury in patients with systemic sclerosis. J. Immunol. 195, 892–900 (2015).
Padilla, C. M. et al. Increased CD8+ tissue resident memory T cells, regulatory T cells, and activated natural killer cells in systemic sclerosis lungs. Rheumatology 63, 837–845 (2023).
Maehara, T. et al. Cytotoxic CD4+ T lymphocytes may induce endothelial cell apoptosis in systemic sclerosis. J. Clin. Invest. 130, 2451–2464 (2020).
Gumkowska-Sroka, O. et al. Cytometric characterization of main immunocompetent cells in patients with systemic sclerosis: relationship with disease activity and type of immunosuppressive treatment. J. Clin. Med. 8, 625 (2019).
Horikawa, M. et al. Abnormal natural killer cell function in systemic sclerosis: altered cytokine production and defective killing activity. J. Invest. Dermatol. 125, 731–737 (2005).
Henriques, A. et al. Subset-specific alterations in frequencies and functional signatures of γδ T cells in systemic sclerosis patients. Inflamm. Res. 65, 985–994 (2016).
Kahaleh, M. B., Fan, P. S. & Otsuka, T. γδ Receptor bearing T cells in scleroderma: enhanced interaction with vascular endothelial cells in vitro. Clin. Immunol. 91, 188–195 (1999).
Schachna, L. et al. Recognition of granzyme B-generated autoantigen fragments in scleroderma patients with ischemic digital loss. Arthritis Rheum. 46, 1873–1884 (2002).
Ulanet, D. B., Flavahan, N. A., Casciola-Rosen, L. & Rosen, A. Selective cleavage of nucleolar autoantigen B23 by granzyme B in differentiated vascular smooth muscle cells: insights into the association of specific autoantibodies with distinct disease phenotypes. Arthritis Rheum. 50, 233–241 (2004).
Overall, C. M. & Dean, R. A. Degradomics: systems biology of the protease web. Pleiotropic roles of MMPs in cancer. Cancer Metastasis Rev. 25, 69–75 (2006).
Sipione, S. et al. Identification of a novel human granzyme B inhibitor secreted by cultured Sertoli cells. J. Immunol. 177, 5051–5058 (2006).
Horvath, A. J. et al. The murine orthologue of human antichymotrypsin: a structural paradigm for clade A3 serpins. J. Biol. Chem. 280, 43168–43178 (2005).
Hsu, I. et al. Serpina3n accelerates tissue repair in a diabetic mouse model of delayed wound healing. Cell Death Dis. 5, e1458 (2014).
Li, F. et al. Neuronal serpina3n is an endogenous protector against blood brain barrier damage following cerebral ischemic stroke. J. Cereb. Blood Flow. Metab. 43, 241–257 (2023).
Haile, Y. et al. Granzyme B-inhibitor serpina3n induces neuroprotection in vitro and in vivo. J. Neuroinflammation 12, 157 (2015).
Saito, A. et al. Blockade of granzyme B remarkably improves mucocutaneous diseases with keratinocyte death in interface dermatitis. J. Invest. Dermatol. 138, 2079–2083 (2018).
Raveney, B. J. E. et al. Eomesodermin-expressing T-helper cells are essential for chronic neuroinflammation. Nat. Commun. 6, 8437 (2015).
Uranga-Murillo, I. et al. Biological relevance of granzymes A and K during E. coli sepsis. Theranostics 11, 9873–9883 (2021).
Sas, G., Pepper, D. S. & Cash, J. D. Investigations on antithrombin III in normal plasma and serum. Br. J. Haematol. 30, 265–272 (1975).
Willoughby, C. A. et al. Discovery of potent, selective human granzyme B inhibitors that inhibit CTL mediated apoptosis. Bioorg. Med. Chem. Lett. 12, 2197–2200 (2002).
Dufour, A. & Overall, C. M. Missing the target: matrix metalloproteinase antitargets in inflammation and cancer. Trends Pharmacol. Sci. 34, 233–242 (2013).
Ebnet, K. et al. Granzyme A-deficient mice retain potent cell-mediated cytotoxicity. EMBO J. 14, 4230–4239 (1995).
Heusel, J. W., Wesselschmidt, R. L., Shresta, S., Russell, J. H. & Ley, T. J. Cytotoxic lymphocytes require granzyme B for the rapid induction of DNA fragmentation and apoptosis in allogeneic target cells. Cell 76, 977–987 (1994).
Joeckel, L. T., Allison, C. C., Pellegrini, M., Bird, C. H. & Bird, P. I. Granzyme K-deficient mice show no evidence of impaired antiviral immunity. Immunol. Cell Biol. 95, 676–683 (2017).
Saito, S., Murakoshi, K., Kotake, S., Kamatani, N. & Tomatsu, T. Granzyme B induces apoptosis of chondrocytes with natural killer cell-like cytotoxicity in rheumatoid arthritis. J. Rheumatol. 35, 1932–1943 (2008).
Zhang, D., Beresford, P. J., Greenberg, A. H. & Lieberman, J. Granzymes A and B directly cleave lamins and disrupt the nuclear lamina during granule-mediated cytolysis. Proc. Natl Acad. Sci. USA 98, 5746–5751 (2001).
Acknowledgements
D.J.G. is funded by grants-in-aid from the Canadian Institutes for Health Research (CIHR). A.A. is the recipient of the Arthritis Society Canada Training Postdoctoral Fellowship. J.P. is funded by CIBER (Centro de Investigación Biomédica en Red; CB21/13/00087), Instituto de Salud Carlos III, FEDER (Fondo Europeo de Desarrollo Regional), Gobierno de Aragón (Group B29_23R, and LMP139_21), Grant PID2020-113963RBI00 by MCIN/AEI/10.13039/501100011033, ASPANOA, and Carrera de la Mujer de Monzón.
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D.J.G. is a co-founder and chief scientific officer of viDA Therapeutics, which owns patents for and is developing inhibitors targeting granzymes as therapeutics. The remaining authors declare no competing interests.
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Aubert, A., Jung, K., Hiroyasu, S. et al. Granzyme serine proteases in inflammation and rheumatic diseases. Nat Rev Rheumatol (2024). https://doi.org/10.1038/s41584-024-01109-5
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DOI: https://doi.org/10.1038/s41584-024-01109-5
