Granzymes (Grs) are serine proteases mainly produced by cytotoxic lymphocytes and are traditionally considered to cause apoptosis in tumor cells and virally infected cells. However, the cytotoxicity of several Grs is currently being debated, and additional, predominantly extracellular, functions of Grs in inflammation are emerging. Extracellular soluble Grs are elevated in the circulation of patients with autoimmune diseases and infections. Additionally, Grs are expressed by several types of immune cells other than cytotoxic lymphocytes. Recent research has revealed novel immunomodulatory functions of Grs. In this review, we provide a comprehensive overview on the role of Grs in inflammation, highlighting their role in cytokine induction and processing.

Our immune defense against tumor cells and virally infected cells is mediated by cytotoxic lymphocytes: NK cells, NKT cells, γδ TCR cells, and cytotoxic T lymphocytes. These cytotoxic cells can induce apoptosis in aberrant cells via the death receptor pathway and the granule exocytosis pathway (1). The death receptor pathway depends on the engagement of death receptors on the target cell by death receptor ligands on the effector cell, whereas the granule exocytosis pathway involves the release of a set of cytotoxic proteins. These are stored in granules in cytotoxic lymphocytes and include the pore-forming protein perforin and a family of structurally related serine proteases called granzymes (Grs) (2, 3). Upon recognition, Grs are released into the immunological synapse between the cytotoxic lymphocyte and the target cell. The Grs then enter the target cell with the aid of perforin and activate various proapoptotic pathways by cleavage of intracellular substrates (1, 4).

In humans, five different Grs exist: GrA, GrB, GrH, GrK, and GrM. This nonsequential nomenclature is explained by the existence of a more extensive Gr family in rodents. Mice express Grs A–G, K, M, and N, but not H, whereas rats express Grs I and J, in addition to Grs A–C, F, K, and M (5). Human Grs (hGrs), named after their rodent homologs, are encoded on three distinct chromosomal regions: GrA and GrK on chromosome 5, GrB and GrH on chromosome 14, and GrM on chromosome 19 (6). Grs A, B, K, and H are expressed in higher quantities in cytotoxic T lymphocytes compared with NK cells, whereas GrM is more abundant in NK cells (1, 2, 4).

Grs are serine proteases that consist of two six-stranded β-barrels that regulate substrate specificity, in the middle of which lies a catalytic triad containing the amino acids serine, histidine, and aspartic acid. Mutation of one of the amino acids in the catalytic triad renders the Gr catalytically inactive. Although human Grs are ∼40% homologous in amino acid sequence (7), they each cleave their own specific set of substrates, a phenomenon partly caused by differences in the primary substrate specificity or P1 of each Gr. GrA and GrK display tryptase-like activity, cleaving their substrates after an arginine or a lysine. Similar to caspases, GrB cleaves substrates after an aspartic acid or glutamic acid. GrM cleaves after a leucine or a methionine, and GrH cleaves after a tyrosine or a phenylalanine (2, 4). Some Grs share substrates, but none have exactly the same degradome. Thus, Grs have substrate specificities that only partially overlap (8). The likeliness that in vitro hydrolysis of a given substrate is physiologically relevant can be estimated by determining the concentration of substrate required for effective hydrolysis (indicated by the Michaelis constant Km, the concentration of enzyme at which the enzyme reaction is at half of its maximum rate) and the time required for the Gr to hydrolyze one substrate molecule (indicated by the apparent unimolecular rate constant or turnover number kcat). When Km is low and kcat is high, the catalysis has a greater probability to be of physiological relevance.

During the past decades, the dogma has been held that all Grs induce cell death. However, this dogma has recently been debated, in particular for GrA and GrK (6, 911). Additionally, increased Gr levels in serum, plasma, synovial fluid, and/or bronchoalveolar lavage fluid have been described in patients suffering from inflammatory diseases, including rheumatoid arthritis (GrA, B), viral infections (GrA, B, K), Plasmodium falciparum infections (GrA, GrB), experimental endotoxemia or sepsis (GrA, GrB, GrK, GrM), hypersensitivity pneumonitis (GrA, GrB), and acute airway inflammation (GrK) (1222). These observations prompted researchers to investigate alternative Gr functions in inflammation. The first evidence for an extracellular role for Grs was provided by Sayers et al. (23), who noted that treatment with extracellular rat Gr causes growth inhibition in tumor cell lines (23). More recently, novel extracellular and perforin-independent functions have been identified (24, 25). It has been demonstrated that Gr-mediated extracellular matrix (ECM) degradation may contribute to inflammation (2630). GrB degrades several ECM components, such as vitronectin, laminin, fibronectin, decorin, and the proteoglycans biglycan and betaglycan (3135). In mouse models, GrB degrades the ECM proteins fibronectin, decorin, fibrillin-1, and vitronectin, and it contributes to several disease processes, including delayed wound closure, abdominal aortic aneurysm, and skin aging (3639). These findings point to a pivotal role of GrB in mediating ECM remodeling in disease. Furthermore, mouse and rat GrA degrade human ECM components (fibronectin and collagen type IV) (4043). However, it is unclear to what extent these findings have physiological relevance. Also, it is unknown whether human GrA, GrH, GrK, and GrM degrade human ECM substrates.

Apart from their role in ECM degradation, a growing body of evidence now implicates GrA, GrB, GrK, and GrM in the production, release, and/or processing of proinflammatory cytokines. This immunomodulatory potential of Grs is discussed in the present review.

Studies in mice show a role for GrA in the inflammatory response to the Gram-negative bacterial cell wall component LPS. GrA−/− mice better withstand a lethal LPS challenge in that they survive longer than do wild-type (WT) mice (11, 44). Furthermore, hGrA exerts direct effects on several cell types to induce cytokine release in vitro. Extracellular GrA stimulates release of proinflammatory cytokines IL-6 from fibroblast cell lines and IL-8 from fibroblast and epithelial cell lines (45). Additionally, extracellular GrA activates primary human monocytes to release IL-1β, TNF-α, IL-6, and IL-8 (11, 46). These effects of GrA are dependent on its catalytic activity, implying that at least one downstream signaling protein needs to be cleaved. The effects of GrA are augmented upon intracellular delivery (11), which suggests that GrA substrates necessary for cytokine induction localize in the cell. Until now, however, these substrates have not been identified. hGrA cleaves and activates pro–IL-1β in vitro and was therefore designated an IL-1β–converting enzyme (47), but the physiological relevance of this finding has never been shown. Moreover, some have failed to demonstrate such an effect of GrA (11). GrA-induced cytokine release from monocytes is blocked in the presence of caspase-1 inhibitors (11), further indicating that IL-1β maturation upon GrA treatment is not mediated by GrA directly, although the inflammasome is involved (Fig. 1). Finally, we have recently found that human recombinant GrA does not induce cytokine release from human monocytes, but synergistically potentiates the LPS-induced cytokine response by these cells (48). This is in line with the observation that mouse macrophages primed with LPS respond to mouse Gr (mGr)A and mGrK (10, 11). We are currently investigating the molecular mechanism underlying this GrA effect. Interestingly, hGrA blocks growth of intracellular mycobacteria in human macrophages (49). Human γδ T cells efficiently inhibit the growth of these bacteria inside macrophages. Recombinant GrA, as well as GrA produced by γδ T cells, induces TNF-α production in macrophages, which in turn inhibits the growth of the pathogen (49). The mechanism by which TNF-α exerts this effect remains unknown. In this regard, it would be interesting to investigate whether GrA, in addition to potentiating TLR4-mediated cytokine responses to LPS (48), also synergizes with other TLR ligands such as mycobacterial products to more efficiently activate monocytes and other innate immune cells.

FIGURE 1.

Overview of effects of Grs on cytokine secretion by different cell types. GrA, GrB, and GrK from human or mouse induce the release of cytokines from several cell types, including fibroblasts (45, 48, 53), monocytes (11, 46, 48), and macrophages (10, 11, 49). Grs cause this release on their own or potentiate LPS-induced cytokine responses. In some cases, catalytically active Gr is required for these effects. This cleavage is indicated by scissors. We have reported Gr functions independent of their catalytic activity (48). Spencer et al. (49) do not report whether GrA activity is required for the observed inhibition of mycobacterial growth.

FIGURE 1.

Overview of effects of Grs on cytokine secretion by different cell types. GrA, GrB, and GrK from human or mouse induce the release of cytokines from several cell types, including fibroblasts (45, 48, 53), monocytes (11, 46, 48), and macrophages (10, 11, 49). Grs cause this release on their own or potentiate LPS-induced cytokine responses. In some cases, catalytically active Gr is required for these effects. This cleavage is indicated by scissors. We have reported Gr functions independent of their catalytic activity (48). Spencer et al. (49) do not report whether GrA activity is required for the observed inhibition of mycobacterial growth.

Close modal

Data on GrB knockout mice survival upon LPS challenge are conflicting. Anthony et al. (44) show that deletion of GrB has no effect on survival upon LPS challenge, whereas Metkar et al. (11) showed that GrB knockout results in a marked increase in survival that exceeds the effect of GrA deletion. Thus, the effect of GrB on the immune response to LPS remains to be elucidated. The reported differences in survival of GrB−/− mice upon LPS challenge may relate to the age of the mice, the injected amount of LPS per gram of body weight, the source of LPS, or other differences in experimental setup.

Interestingly, Metkar et al. (11) reported a worse survival of GrAB double knockout mice compared with GrA or GrB single knockout mice upon LPS challenge (11). Anthony et al. (44), however, found no difference in survival of GrA−/− mice and GrAB−/− mice after LPS injection, consistent with their finding that GrB does not influence survival. GrB−/− and GrAB−/− mice were not directly compared in their experiments (44). The interplay between GrA and GrB in the immune response to LPS is unresolved and deserves further study. Possibly, different Grs are preferentially involved in response to different types of LPS.

In vitro evidence that GrB is involved in cytokine release during inflammatory responses is scarce. hGrB itself does not directly induce HeLa and HUVEC cells to produce IL-6 and IL-8 (50). However, we recently demonstrated that hGrB synergistically enhances LPS-induced TNF-α release from human monocytes in vitro, but it does not induce cytokine release from these cells on its own (48). This function of hGrB is shared with at least hGrK and hGrA (48), a redundancy consistent with a limited effect of mGrB on survival in response to LPS (44). A role for human GrB in the processing of proinflammatory cytokines also has been described. GrB cleaves pro–IL-18 in vitro and ex vivo, at the same position as caspase-1, although with slower kinetics (51, 52). Furthermore, hGrB cleaves the 31-kDa precursor of IL-1α, which enhances the biological activity of the cytokine several fold, an in vitro result that was confirmed in mice (50). Thus, it appears that extracellular GrB is able to cleave and activate several important proinflammatory cytokines. In conclusion, GrB may have more effect on cytokine cleavage and activation than on initial cytokine release.

Mice deficient in GrK have not been described yet. Hence, it is currently unknown whether GrK deficiency in mice results in altered responses to LPS challenges. The effects of GrK on different cell types in vitro, however, have been investigated. Similar to GrA (45), extracellular hGrK releases IL-6 and IL-8 from human lung fibroblasts, dependent on GrK proteolytic activity (48, 53). This effect of GrK involves cleavage of protease-activated receptor 1 (53). Additionally, recombinant mGrK, but not its catalytically inactive proform, induces release of IL-1β from peritoneal mouse macrophages (10), an effect also observed for mGrA (11). This effect is seen after priming the cells with LPS, which may indicate an additive or synergistic effect of Grs and bacterial compounds. Intracellular GrK delivery is required, although extracellular effects are observed at concentrations >600 nM (53). Cytotoxic T cells isolated from lymphocytic choriomeningitis virus–infected GrA/B-deficient mice, which mainly expressed GrK, also induce IL-1β release from mouse macrophages (10). We have recently shown that extracellular human GrK synergistically potentiates LPS-induced release of the proinflammatory cytokines TNF-α, IL-6, and IL-8 in human monocytes in vitro (48). Additionally, a combination of GrK and LPS enhances release of TNF-α, IL-1β and IL-6 in vivo, compared with LPS alone. This effect is independent of GrK proteolytic activity. GrK liberates individual LPS molecules from micelles and promotes complex formation between LPS and CD14 (48). Taken together, these data indicate that the proinflammatory effects of GrK are diverse and cell type specific. The proinflammatory effects of GrA, GrB, and GrK on different cell types are summarized in Fig. 1.

Upon LPS challenge, GrM−/− mice survive longer and produce significantly lower serum levels of the proinflammatory cytokines IL-1α, IL-1β, TNF, and IFN-γ, compared with WT mice (44). Perforin knockout partially shows the same phenotype as GrM knockout (44), indicating that Grs may function intracellularly in the LPS response. However, recombinant mGrM does not cleave pro–IL-1β in vitro, and the molecular role of GrM in promoting responsiveness to an endotoxin challenge remains elusive (44). Consistent with results obtained with single knockout mice, GrM/GrA double knockout mice are even more resistant to LPS than are single GrM or GrA knockout mice (44). This additional effect of a double Gr knockout probably indicates that GrM and GrA function in the immune response to LPS via different mechanisms.

GrM colocalizes with the chemotactic protein MIP-1α in cytotoxic vesicles of human NK cells (54). GrM knockout leads to impaired MIP-1α secretion from NK cells and macrophages isolated from the liver of mice challenged with LPS or the Gram-positive bacterium Listeria monocytogenes (54). Because MIP-1α is important for NK cell recruitment to the liver during infection, the authors propose that GrM regulates this process (54). Similar to the response upon LPS challenge, GrM−/− mice survive longer and exhibit reduced serum levels of cytokines following L. monocytogenes infection (54). Interestingly, however, MIP-1α serum levels were not different between WT and GrM−/− serum levels after L. monocytogenes infection (54). Apparently, the local effect of GrM knockout on MIP-1α secretion is not reflected systemically. Taken together, GrM appears to be an important regulator of cytokine release in response to infection with Gram-negative as well as Gram-positive bacteria, at least in mice. However, in vitro data explaining the mechanism(s) behind these observations are lacking.

It is not known how Grs are released during inflammation. Leakage of Grs out of the immunological synapse during ongoing cytotoxic lymphocyte responses could explain their extracellular presence. Alternatively, Grs could be actively secreted during inflammation. GrA and GrB are released upon LPS injection into healthy human volunteers and upon incubation of whole-blood cultures with LPS or bacteria (19), suggesting that purposeful degranulation may occur in the absence of cytotoxicity. Alternatively, secretion of Grs may take place independently from degranulation, because significant amounts of GrA and GrB continue to be secreted from cytotoxic lymphocytes in the absence of continued stimulation by antigenic cells (55).

The source of extracellular Grs is also unknown. Cytotoxic lymphocytes may evidently produce these extracellular Grs. Anthony et al. (44) observed that NK cell depletion, similar to GrM deletion in WT mice, confers LPS resistance to RAG-1−/− mice. Thus, NK cell–derived mGrM may play an important role in the inflammatory response to LPS (44). However, alternative sources of Grs also have to be considered. There is ample evidence that both human myeloid dendritic cells (mDCs) and human plasmacytoid DCs (pDCs) express GrB (5660). Constitutive mRNA and protein expression has been found in pDCs (57, 60), whereas IL-3, IL-10, and IL-21 upregulate GrB mRNA and protein expression in these cells (57, 58, 60). In mDCs, no constitutive GrB protein expression is reported, but GrB and perforin protein expression can be induced by treatment with TLR7 and TLR8 agonists (59). In contrast, TLR7 and TLR9 agonists inhibit GrB protein expression in pDCs (5760).

GrB expression by DCs may contribute to their cytotoxic potential in perforin-dependent as well as perforin-independent ways (61). Human mDCs upregulate perforin protein expression upon stimulation with TLR7 and/or TLR8 ligands and release perforin together with GrB (59). However, no such upregulation is found in pDCs stimulated with TLR7/8 ligands (59) or IL-3/IL-10 (58). GrB-expressing mDCs (59) and pDCs (56, 57, 59) eliminate endothelial cells and tumor cell lines. This cytotoxic activity may provide DCs with extra possibilities to regulate inflammatory processes. Interestingly, human immature DCs eliminate CD8+ T cells via a perforin- and GrA-dependent mechanism (62). Gr expression could therefore depend on the developmental stage of the DC, because mature pDCs do not express GrA (57). Human B cells also express GrB (6365), and as for DCs, GrB expression is induced by IL-21, albeit in the context of other stimuli, including viral Ags (65). GrB from B cells was shown to inhibit T cell proliferation via the cleavage of the TCR ζ-chain (63), an extracellular and perforin-independent mechanism also shown for GrB derived from cytotoxic lymphocytes (66). Furthermore, GrB-secreting B cells possess cytotoxic potential in the absence of perforin expression (64), which is thought to contribute to their immunoregulatory potential (66). In contrast, Hagn et al. (67) could not detect GrB expression in resting or activated murine B cells.

GrB protein expression has also been found in human macrophages (68), basophils (69), mast cells (70), and several other cell types (66). GrB expression in mast cells and basophils was not accompanied by perforin expression (69, 70), indicating perforin-independent and possibly extracellular functions for GrB released by these cells. Further studies are required regarding the cellular origin of Gr in individual diseases, the regulation of Gr release, and the molecular immunoregulatory mechanisms Grs may employ.

Another issue to be clarified is whether Grs stimulate the release of previously synthesized cytokines stored inside the cell, or whether they exert a stimulatory effect on cytokine mRNA transcription and protein synthesis, or both. Not much is known about the pathways that are activated after extracellular Gr treatment. GrA can be internalized by monocytes in the absence of perforin, and it induces cytokine release (11), but the link between these events is unclear. Upon cleavage of protease-activated receptor 1 by GrK, ERK1/2 and p38 MAPK are activated (53), which then leads to cytokine release from fibroblasts. More research is needed to clarify which signaling pathways are activated by Grs to trigger cytokine responses. When Gr catalytic activity is required, identification of intra- or extracellular Gr substrates is essential. In cases where Grs potentiate pathways induced by TLR ligands independent of the Gr catalytic activity (48), the focus should lie on clarifying how Grs modulate these pathways via interaction with signaling molecules. A common characteristic in LPS-enhancing proteins such as LPS-binding protein (71, 72), Grs (48), the neutrophil granule protein azurocidin (73, 74), high mobility group box-1 protein (75, 76), protamines (77), and apolipoprotein C1 (78, 79) is the presence of cationic patches of arginines and/or lysines that drive the interaction with LPS. Endocytosis may also play a role, because internalization is required for the stimulating effect of azurocidin on LPS-induced cytokine responses (80). Interestingly, GrA binds to and is internalized by monocytes (11).

Another question is whether extracellular circulating Grs are active, or whether their activity is reduced due to binding by protease inhibitors. In vivo, the catalytic activity of Grs and other serine proteases is inhibited by serine protease inhibitors or serpins (81). Inhibition occurs through proteolytic attack of the serpin by the Grs, after which a covalent bond is formed between the serpin and the active site of Gr (81). Enzyme/serpin complexes are subsequently removed from the circulation by the liver. Removal is rapid and often occurs within minutes after complex formation (82, 83).

Several endogenous serpins for human Grs have been identified. Furthermore, several general protease inhibitors with the potential to inhibit the catalytic activity of Grs are present in the human circulation. GrA is inhibited by the serpin Kazal (84) and extracellular GrA is inhibited by anti-thrombin III and α2-macroglobulin (85). The Gr is protected from inhibition when it is complexed to proteoglycans (85). GrB is inhibited by serpin B9, also known as proteinase inhibitor 9 (PI-9) (86). PI-9 is thought to operate intracellularly, but is also present in the circulation where it is complexed to GrB (87). Still, GrB activity is retained in the presence of 80% plasma (88), and therefore it is unclear whether PI-9 inhibits GrB activity in blood. GrB catalytic activity is not reduced by α1-antitrypsin (88). The three major extracellular protease inhibitors present in normal lung fluid (α1-antitrypsin, elafin, and secretory leukocyte protease inhibitor) do not inhibit GrA or GrB, and GrA is active in bronchoalveolar lavage fluid (20). GrH is inhibited by serpin B1 (89), and GrM is inhibited by serpin B4 (90). Furthermore, GrM is inhibited by α1-antichymotrypsin and α1-proteinase inhibitor, although weakly (91). GrK catalytic activity is inhibited by inter–α-trypsin inhibitor (92). In sepsis patients, circulating amounts of inter–α-trypsin inhibitor are lower than in healthy controls (15, 93) Also, the molecular mass of GrK in the serum of sepsis patients is lower than in healthy controls. GrK mainly circulates with a molecular mass of ∼26 kDa in sepsis patients (15). This finding implies that GrK is freed from protease inhibitors under inflammatory conditions, although it was not determined whether this GrK is proteolytically active.

In summary, inhibition of catalytic activity of Grs may be reduced under inflammatory conditions, although this subject requires further study. Apart from intracellular inhibition of GrB by PI-9, the physiological relevance of serpins or other protease inhibitors to inhibit Gr activity remains unknown. Furthermore, it has to be considered that proteolytic activity may not be essential for all extracellular Gr functions (48). In this regard, steric hindrance by binding of inhibitors or proteoglycans could be of superior importance.

Increasing evidence now demonstrates that Grs are involved in ECM remodeling and modulation of inflammatory pathways. Grs may induce cytokine release from cells and/or cleave cytokines after cytokine release. Involvement of GrA, GrB, GrK, and GrM in these processes has been demonstrated in humans and/or mice (11, 44, 4850, 53, 54). Most of these novel Gr functions are perforin-independent and extracellular, although effects may be enhanced upon intracellular delivery. Furthermore, Grs have immunomodulatory functions independent of their catalytic activity (48). Taken together, these data indicate that Grs have multiple regulatory functions in (innate) proinflammatory immune responses via many at least partly overlapping pathways (Fig. 1). This functional redundancy underlines the significance of Grs in immunity and ensures proper Gr function when one of the Grs or Gr-induced mechanisms is inhibited. The notion that multiple Grs influence inflammatory processes is reminiscent of the cytotoxic potential shared by several Grs, which is regarded as a safeguard against inhibition of one or more Grs (4).

Several important questions remain to be solved. First, not all Grs have been studied in the context of inflammation. It is unknown whether GrH and GrM possess direct cytokine-inducing and/or -processing capacity. Second, in many cases the intracellular mechanisms behind the effects of Grs on cytokine secretion have not been elucidated. Third, it is not understood how Gr secretion is regulated under inflammatory conditions and from which cell type(s) Grs originate. Finally, the regulation of Gr activity in the circulation also deserves further study. Elucidation of all these different aspects of Gr biology will help us to better understand the implications of Gr activity in disease. Once clear molecular mechanisms have been defined for Grs in inflammatory disease, and their physiological relevance has been demonstrated, Gr inhibition might in the future contribute to treatment of infectious diseases and autoimmune diseases.

This work was supported by the University Medical Center Utrecht (to C.E.H.) and by Netherlands Organization for Scientific Research Grant 916.66.044 (to N.B.).

Abbreviations used in this article:

DC

dendritic cell

ECM

extracellular matrix

Gr

granzyme

hGr

human Gr

mDC

myeloid dendritic cell

mGr

mouse Gr

pDC

plasmacytoid DC

PI-9

proteinase inhibitor 9

WT

wild-type.

1
Anthony
D. A.
,
Andrews
D. M.
,
Watt
S. V.
,
Trapani
J. A.
,
Smyth
M. J.
.
2010
.
Functional dissection of the granzyme family: cell death and inflammation.
Immunol. Rev.
235
:
73
92
.
2
Bovenschen
N.
,
Kummer
J. A.
.
2010
.
Orphan granzymes find a home.
Immunol. Rev.
235
:
117
127
.
3
de Poot
S. A.
,
Bovenschen
N.
.
2014
.
Granzyme M: behind enemy lines.
Cell Death Differ.
21
:
359
368
.
4
Chowdhury
D.
,
Lieberman
J.
.
2008
.
Death by a thousand cuts: granzyme pathways of programmed cell death.
Annu. Rev. Immunol.
26
:
389
420
.
5
Susanto
O.
,
Trapani
J. A.
,
Brasacchio
D.
.
2012
.
Controversies in granzyme biology.
Tissue Antigens
80
:
477
487
.
6
Joeckel
L. T.
,
Bird
P. I.
.
2014
.
Are all granzymes cytotoxic in vivo?
Biol. Chem.
395
:
181
202
.
7
Sattar
R.
,
Ali
S. A.
,
Abbasi
A.
.
2003
.
Bioinformatics of granzymes: sequence comparison and structural studies on granzyme family by homology modeling.
Biochem. Biophys. Res. Commun.
308
:
726
735
.
8
Bovenschen
N.
,
Quadir
R.
,
van den Berg
A. L.
,
Brenkman
A. B.
,
Vandenberghe
I.
,
Devreese
B.
,
Joore
J.
,
Kummer
J. A.
.
2009
.
Granzyme K displays highly restricted substrate specificity that only partially overlaps with granzyme A.
J. Biol. Chem.
284
:
3504
3512
.
9
Kaiserman
D.
,
Stewart
S. E.
,
Plasman
K.
,
Gevaert
K.
,
Van Damme
P.
,
Bird
P. I.
.
2014
.
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
.
10
Joeckel
L. T.
,
Wallich
R.
,
Martin
P.
,
Sanchez-Martinez
D.
,
Weber
F. C.
,
Martin
S. F.
,
Borner
C.
,
Pardo
J.
,
Froelich
C.
,
Simon
M. M.
.
2011
.
Mouse granzyme K has pro-inflammatory potential.
Cell Death Differ.
18
:
1112
1119
.
11
Metkar
S. S.
,
Menaa
C.
,
Pardo
J.
,
Wang
B.
,
Wallich
R.
,
Freudenberg
M.
,
Kim
S.
,
Raja
S. M.
,
Shi
L.
,
Simon
M. M.
,
Froelich
C. J.
.
2008
.
Human and mouse granzyme A induce a proinflammatory cytokine response.
Immunity
29
:
720
733
.
12
Bovenschen
N.
,
Spijkers
S. N.
,
Wensink
A. C.
,
Schellens
I. M.
,
van Domselaar
R.
,
van Baarle
D.
.
2014
.
Elevated granzyme M-expressing lymphocytes during cytomegalovirus latency and reactivation after allogeneic stem cell transplantation.
Clin. Immunol.
150
:
1
11
.
13
Hollestelle
M. J.
,
Lai
K. W.
,
van Deuren
M.
,
Lenting
P. J.
,
de Groot
P. G.
,
Sprong
T.
,
Bovenschen
N.
.
2011
.
Cleavage of von Willebrand factor by granzyme M destroys its factor VIII binding capacity.
PLoS ONE
6
:
e24216
.
14
Bratke
K.
,
Klug
A.
,
Julius
P.
,
Kuepper
M.
,
Lommatzsch
M.
,
Sparmann
G.
,
Luttmann
W.
,
Virchow
J. C.
.
2008
.
Granzyme K: a novel mediator in acute airway inflammation.
Thorax
63
:
1006
1011
.
15
Rucevic
M.
,
Fast
L. D.
,
Jay
G. D.
,
Trespalcios
F. M.
,
Sucov
A.
,
Siryaporn
E.
,
Lim
Y. P.
.
2007
.
Altered levels and molecular forms of granzyme K in plasma from septic patients.
Shock
27
:
488
493
.
16
Bade
B.
,
Lohrmann
J.
,
ten Brinke
A.
,
Wolbink
A. M.
,
Wolbink
G. J.
,
ten Berge
I. J.
,
Virchow
J. C.
 Jr.
,
Luttmann
W.
,
Hack
C. E.
.
2005
.
Detection of soluble human granzyme K in vitro and in vivo.
Eur. J. Immunol.
35
:
2940
2948
.
17
Zeerleder
S.
,
Hack
C. E.
,
Caliezi
C.
,
van Mierlo
G.
,
Eerenberg-Belmer
A.
,
Wolbink
A.
,
Wuillenmin
W. A.
.
2005
.
Activated cytotoxic T cells and NK cells in severe sepsis and septic shock and their role in multiple organ dysfunction.
Clin. Immunol.
116
:
158
165
.
18
Hermsen
C. C.
,
Konijnenberg
Y.
,
Mulder
L.
,
Loé
C.
,
van Deuren
M.
,
van der Meer
J. W.
,
van Mierlo
G. J.
,
Eling
W. M.
,
Hack
C. E.
,
Sauerwein
R. W.
.
2003
.
Circulating concentrations of soluble granzyme A and B increase during natural and experimental Plasmodium falciparum infections.
Clin. Exp. Immunol.
132
:
467
472
.
19
Lauw
F. N.
,
Simpson
A. J.
,
Hack
C. E.
,
Prins
J. M.
,
Wolbink
A. M.
,
van Deventer
S. J.
,
Chaowagul
W.
,
White
N. J.
,
van Der Poll
T.
.
2000
.
Soluble granzymes are released during human endotoxemia and in patients with severe infection due to Gram-negative bacteria.
J. Infect. Dis.
182
:
206
213
.
20
Tremblay
G. M.
,
Wolbink
A. M.
,
Cormier
Y.
,
Hack
C. E.
.
2000
.
Granzyme activity in the inflamed lung is not controlled by endogenous serine proteinase inhibitors.
J. Immunol.
165
:
3966
3969
.
21
Tak
P. P.
,
Spaeny-Dekking
L.
,
Kraan
M. C.
,
Breedveld
F. C.
,
Froelich
C. J.
,
Hack
C. E.
.
1999
.
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
.
22
Spaeny-Dekking
E. H.
,
Hanna
W. L.
,
Wolbink
A. M.
,
Wever
P. C.
,
Kummer
J. A.
,
Swaak
A. J.
,
Middeldorp
J. M.
,
Huisman
H. G.
,
Froelich
C. J.
,
Hack
C. E.
.
1998
.
Extracellular granzymes A and B in humans: detection of native species during CTL responses in vitro and in vivo.
J. Immunol.
160
:
3610
3616
.
23
Sayers
T. J.
,
Wiltrout
T. A.
,
Sowder
R.
,
Munger
W. L.
,
Smyth
M. J.
,
Henderson
L. E.
.
1992
.
Purification of a factor from the granules of a rat natural killer cell line (RNK) that reduces tumor cell growth and changes tumor morphology. Molecular identity with a granule serine protease (RNKP-1).
J. Immunol.
148
:
292
300
.
24
van Domselaar
R.
,
Bovenschen
N.
.
2011
.
Cell death-independent functions of granzymes: hit viruses where it hurts.
Rev. Med. Virol.
21
:
301
314
.
25
Andrade
F.
2010
.
Non-cytotoxic antiviral activities of granzymes in the context of the immune antiviral state.
Immunol. Rev.
235
:
128
146
.
26
Hiebert
P. R.
,
Granville
D. J.
.
2012
.
Granzyme B in injury, inflammation, and repair.
Trends Mol. Med.
18
:
732
741
.
27
Hendel
A.
,
Hiebert
P. R.
,
Boivin
W. A.
,
Williams
S. J.
,
Granville
D. J.
.
2010
.
Granzymes in age-related cardiovascular and pulmonary diseases.
Cell Death Differ.
17
:
596
606
.
28
Sorokin
L.
2010
.
The impact of the extracellular matrix on inflammation.
Nat. Rev. Immunol.
10
:
712
723
.
29
Boivin
W. A.
,
Cooper
D. M.
,
Hiebert
P. R.
,
Granville
D. J.
.
2009
.
Intracellular versus extracellular granzyme B in immunity and disease: challenging the dogma.
Lab. Invest.
89
:
1195
1220
.
30
Buzza
M. S.
,
Bird
P. I.
.
2006
.
Extracellular granzymes: current perspectives.
Biol. Chem.
387
:
827
837
.
31
Boivin
W. A.
,
Shackleford
M.
,
Vanden Hoek
A.
,
Zhao
H.
,
Hackett
T. L.
,
Knight
D. A.
,
Granville
D. J.
.
2012
.
Granzyme B cleaves decorin, biglycan and soluble betaglycan, releasing active transforming growth factor-β1.
PLoS ONE
7
:
e33163
.
32
Buzza
M. S.
,
Zamurs
L.
,
Sun
J.
,
Bird
C. H.
,
Smith
A. I.
,
Trapani
J. A.
,
Froelich
C. J.
,
Nice
E. C.
,
Bird
P. I.
.
2005
.
Extracellular matrix remodeling by human granzyme B via cleavage of vitronectin, fibronectin, and laminin.
J. Biol. Chem.
280
:
23549
23558
.
33
Choy
J. C.
,
Hung
V. H.
,
Hunter
A. L.
,
Cheung
P. K.
,
Motyka
B.
,
Goping
I. S.
,
Sawchuk
T.
,
Bleackley
R. C.
,
Podor
T. J.
,
McManus
B. M.
,
Granville
D. J.
.
2004
.
Granzyme B induces smooth muscle cell apoptosis in the absence of perforin: involvement of extracellular matrix degradation.
Arterioscler. Thromb. Vasc. Biol.
24
:
2245
2250
.
34
Ronday
H. K.
,
van der Laan
W. H.
,
Tak
P. P.
,
de Roos
J. A.
,
Bank
R. A.
,
TeKoppele
J. M.
,
Froelich
C. J.
,
Hack
C. E.
,
Hogendoorn
P. C.
,
Breedveld
F. C.
,
Verheijen
J. H.
.
2001
.
Human granzyme B mediates cartilage proteoglycan degradation and is expressed at the invasive front of the synovium in rheumatoid arthritis.
Rheumatology (Oxford)
40
:
55
61
.
35
Froelich
C. J.
,
Zhang
X.
,
Turbov
J.
,
Hudig
D.
,
Winkler
U.
,
Hanna
W. L.
.
1993
.
Human granzyme B degrades aggrecan proteoglycan in matrix synthesized by chondrocytes.
J. Immunol.
151
:
7161
7171
.
36
Hiebert
P. R.
,
Wu
D.
,
Granville
D. J.
.
2013
.
Granzyme B degrades extracellular matrix and contributes to delayed wound closure in apolipoprotein E knockout mice.
Cell Death Differ.
20
:
1404
1414
.
37
Hiebert
P. R.
,
Boivin
W. A.
,
Abraham
T.
,
Pazooki
S.
,
Zhao
H.
,
Granville
D. J.
.
2011
.
Granzyme B contributes to extracellular matrix remodeling and skin aging in apolipoprotein E knockout mice.
Exp. Gerontol.
46
:
489
499
.
38
Ang
L. S.
,
Boivin
W. A.
,
Williams
S. J.
,
Zhao
H.
,
Abraham
T.
,
Carmine-Simmen
K.
,
McManus
B. M.
,
Bleackley
R. C.
,
Granville
D. J.
.
2011
.
Serpina3n attenuates granzyme B-mediated decorin cleavage and rupture in a murine model of aortic aneurysm.
Cell Death Dis.
2
:
e209
.
39
Chamberlain
C. M.
,
Ang
L. S.
,
Boivin
W. A.
,
Cooper
D. M.
,
Williams
S. J.
,
Zhao
H.
,
Hendel
A.
,
Folkesson
M.
,
Swedenborg
J.
,
Allard
M. F.
, et al
.
2010
.
Perforin-independent extracellular granzyme B activity contributes to abdominal aortic aneurysm.
Am. J. Pathol.
176
:
1038
1049
.
40
Hirayasu
H.
,
Yoshikawa
Y.
,
Tsuzuki
S.
,
Fushiki
T.
.
2008
.
A lymphocyte serine protease granzyme A causes detachment of a small-intestinal epithelial cell line (IEC-6).
Biosci. Biotechnol. Biochem.
72
:
2294
2302
.
41
Vettel
U.
,
Brunner
G.
,
Bar-Shavit
R.
,
Vlodavsky
I.
,
Kramer
M. D.
.
1993
.
Charge-dependent binding of granzyme A (MTSP-1) to basement membranes.
Eur. J. Immunol.
23
:
279
282
.
42
Simon
M. M.
,
Kramer
M. D.
,
Prester
M.
,
Gay
S.
.
1991
.
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
.
43
Simon
M. M.
,
Prester
M.
,
Nerz
G.
,
Kramer
M. D.
,
Fruth
U.
.
1988
.
Release of biologically active fragments from human plasma-fibronectin by murine T cell-specific proteinase 1 (TSP-1).
Biol. Chem. Hoppe Seyler
369
(
Suppl
):
107
112
.
44
Anthony
D. A.
,
Andrews
D. M.
,
Chow
M.
,
Watt
S. V.
,
House
C.
,
Akira
S.
,
Bird
P. I.
,
Trapani
J. A.
,
Smyth
M. J.
.
2010
.
A role for granzyme M in TLR4-driven inflammation and endotoxicosis.
J. Immunol.
185
:
1794
1803
.
45
Sower
L. E.
,
Klimpel
G. R.
,
Hanna
W.
,
Froelich
C. J.
.
1996
.
Extracellular activities of human granzymes. I. Granzyme A induces IL6 and IL8 production in fibroblast and epithelial cell lines.
Cell. Immunol.
171
:
159
163
.
46
Sower
L. E.
,
Froelich
C. J.
,
Allegretto
N.
,
Rose
P. M.
,
Hanna
W. D.
,
Klimpel
G. R.
.
1996
.
Extracellular activities of human granzyme A. Monocyte activation by granzyme A versus alpha-thrombin.
J. Immunol.
156
:
2585
2590
.
47
Irmler
M.
,
Hertig
S.
,
MacDonald
H. R.
,
Sadoul
R.
,
Becherer
J. D.
,
Proudfoot
A.
,
Solari
R.
,
Tschopp
J.
.
1995
.
Granzyme A is an interleukin 1 beta-converting enzyme.
J. Exp. Med.
181
:
1917
1922
.
48
Wensink
A. C.
,
Kemp
V.
,
Fermie
J.
,
García Laorden
M. I.
,
van der Poll
T.
,
Hack
C. E.
,
Bovenschen
N.
.
2014
.
Granzyme K synergistically potentiates LPS-induced cytokine responses in human monocytes.
Proc. Natl. Acad. Sci. USA
111
:
5974
5979
.
49
Spencer
C. T.
,
Abate
G.
,
Sakala
I. G.
,
Xia
M.
,
Truscott
S. M.
,
Eickhoff
C. S.
,
Linn
R.
,
Blazevic
A.
,
Metkar
S. S.
,
Peng
G.
, et al
.
2013
.
Granzyme A produced by γ9δ2 T cells induces human macrophages to inhibit growth of an intracellular pathogen.
PLoS Pathog.
9
:
e1003119
.
50
Afonina
I. S.
,
Tynan
G. A.
,
Logue
S. E.
,
Cullen
S. P.
,
Bots
M.
,
Lüthi
A. U.
,
Reeves
E. P.
,
McElvaney
N. G.
,
Medema
J. P.
,
Lavelle
E. C.
,
Martin
S. J.
.
2011
.
Granzyme B-dependent proteolysis acts as a switch to enhance the proinflammatory activity of IL-1α.
Mol. Cell
44
:
265
278
.
51
Akeda
T.
,
Yamanaka
K.
,
Tsuda
K.
,
Omoto
Y.
,
Gabazza
E. C.
,
Mizutani
H.
.
2014
.
CD8+ T cell granzyme B activates keratinocyte endogenous IL-18.
Arch. Dermatol. Res.
306
:
125
130
.
52
Omoto
Y.
,
Yamanaka
K.
,
Tokime
K.
,
Kitano
S.
,
Kakeda
M.
,
Akeda
T.
,
Kurokawa
I.
,
Gabazza
E. C.
,
Tsutsui
H.
,
Katayama
N.
, et al
.
2010
.
Granzyme B is a novel interleukin-18 converting enzyme.
J. Dermatol. Sci.
59
:
129
135
.
53
Cooper
D. M.
,
Pechkovsky
D. V.
,
Hackett
T. L.
,
Knight
D. A.
,
Granville
D. J.
.
2011
.
Granzyme K activates protease-activated receptor-1.
PLoS ONE
6
:
e21484
.
54
Baschuk
N.
,
Wang
N.
,
Watt
S. V.
,
Halse
H.
,
House
C.
,
Bird
P. I.
,
Strugnell
R.
,
Trapani
J. A.
,
Smyth
M. J.
,
Andrews
D. M.
.
2014
.
NK cell intrinsic regulation of MIP-1α by granzyme M.
Cell Death Dis.
5
:
e1115
.
55
Isaaz
S.
,
Baetz
K.
,
Olsen
K.
,
Podack
E.
,
Griffiths
G. M.
.
1995
.
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
.
56
Manna
P. P.
,
Hira
S. K.
,
Das
A. A.
,
Bandyopadhyay
S.
,
Gupta
K. K.
.
2013
.
IL-15 activated human peripheral blood dendritic cell kill allogeneic and xenogeneic endothelial cells via apoptosis.
Cytokine
61
:
118
126
.
57
Bratke
K.
,
Nielsen
J.
,
Manig
F.
,
Klein
C.
,
Kuepper
M.
,
Geyer
S.
,
Julius
P.
,
Lommatzsch
M.
,
Virchow
J. C.
.
2010
.
Functional expression of granzyme B in human plasmacytoid dendritic cells: a role in allergic inflammation.
Clin. Exp. Allergy
40
:
1015
1024
.
58
Jahrsdörfer
B.
,
Vollmer
A.
,
Blackwell
S. E.
,
Maier
J.
,
Sontheimer
K.
,
Beyer
T.
,
Mandel
B.
,
Lunov
O.
,
Tron
K.
,
Nienhaus
G. U.
, et al
.
2010
.
Granzyme B produced by human plasmacytoid dendritic cells suppresses T-cell expansion.
Blood
115
:
1156
1165
.
59
Stary
G.
,
Bangert
C.
,
Tauber
M.
,
Strohal
R.
,
Kopp
T.
,
Stingl
G.
.
2007
.
Tumoricidal activity of TLR7/8-activated inflammatory dendritic cells.
J. Exp. Med.
204
:
1441
1451
.
60
Karrich
J. J.
,
Jachimowski
L. C.
,
Nagasawa
M.
,
Kamp
A.
,
Balzarolo
M.
,
Wolkers
M. C.
,
Uittenbogaart
C. H.
,
Marieke van Ham
S.
,
Blom
B.
.
2013
.
IL-21-stimulated human plasmacytoid dendritic cells secrete granzyme B, which impairs their capacity to induce T-cell proliferation.
Blood
121
:
3103
3111
.
61
Tel
J.
,
Anguille
S.
,
Waterborg
C. E.
,
Smits
E. L.
,
Figdor
C. G.
,
de Vries
I. J.
.
2014
.
Tumoricidal activity of human dendritic cells.
Trends Immunol.
35
:
38
46
.
62
Zangi
L.
,
Klionsky
Y. Z.
,
Yarimi
L.
,
Bachar-Lustig
E.
,
Eidelstein
Y.
,
Shezen
E.
,
Hagin
D.
,
Ito
Y.
,
Takai
T.
,
Reich-Zeliger
S.
, et al
.
2012
.
Deletion of cognate CD8 T cells by immature dendritic cells: a novel role for perforin, granzyme A, TREM-1, and TLR7.
Blood
120
:
1647
1657
.
63
Lindner
S.
,
Dahlke
K.
,
Sontheimer
K.
,
Hagn
M.
,
Kaltenmeier
C.
,
Barth
T. F.
,
Beyer
T.
,
Reister
F.
,
Fabricius
D.
,
Lotfi
R.
, et al
.
2013
.
Interleukin 21-induced granzyme B-expressing B cells infiltrate tumors and regulate T cells.
Cancer Res.
73
:
2468
2479
.
64
Hagn
M.
,
Sontheimer
K.
,
Dahlke
K.
,
Brueggemann
S.
,
Kaltenmeier
C.
,
Beyer
T.
,
Hofmann
S.
,
Lunov
O.
,
Barth
T. F.
,
Fabricius
D.
, et al
.
2012
.
Human B cells differentiate into granzyme B-secreting cytotoxic B lymphocytes upon incomplete T-cell help.
Immunol. Cell Biol.
90
:
457
467
.
65
Hagn
M.
,
Schwesinger
E.
,
Ebel
V.
,
Sontheimer
K.
,
Maier
J.
,
Beyer
T.
,
Syrovets
T.
,
Laumonnier
Y.
,
Fabricius
D.
,
Simmet
T.
,
Jahrsdörfer
B.
.
2009
.
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
.
66
Hagn
M.
,
Jahrsdörfer
B.
.
2012
.
Why do human B cells secrete granzyme B? Insights into a novel B-cell differentiation pathway.
OncoImmunology
1
:
1368
1375
.
67
Hagn
M.
,
Belz
G. T.
,
Kallies
A.
,
Sutton
V. R.
,
Thia
K. Y.
,
Tarlinton
D. M.
,
Hawkins
E. D.
,
Trapani
J. A.
.
2012
.
Activated mouse B cells lack expression of granzyme B.
J. Immunol.
188
:
3886
3892
.
68
Kim
W. J.
,
Kim
H.
,
Suk
K.
,
Lee
W. H.
.
2007
.
Macrophages express granzyme B in the lesion areas of atherosclerosis and rheumatoid arthritis.
Immunol. Lett.
111
:
57
65
.
69
Tschopp
C. M.
,
Spiegl
N.
,
Didichenko
S.
,
Lutmann
W.
,
Julius
P.
,
Virchow
J. C.
,
Hack
C. E.
,
Dahinden
C. A.
.
2006
.
Granzyme B, a novel mediator of allergic inflammation: its induction and release in blood basophils and human asthma.
Blood
108
:
2290
2299
.
70
Strik
M. C.
,
de Koning
P. J.
,
Kleijmeer
M. J.
,
Bladergroen
B. A.
,
Wolbink
A. M.
,
Griffith
J. M.
,
Wouters
D.
,
Fukuoka
Y.
,
Schwartz
L. B.
,
Hack
C. E.
, et al
.
2007
.
Human mast cells produce and release the cytotoxic lymphocyte associated protease granzyme B upon activation.
Mol. Immunol.
44
:
3462
3472
.
71
Eckert
J. K.
,
Kim
Y. J.
,
Kim
J. I.
,
Gürtler
K.
,
Oh
D. Y.
,
Sur
S.
,
Lundvall
L.
,
Hamann
L.
,
van der Ploeg
A.
,
Pickkers
P.
, et al
.
2013
.
The crystal structure of lipopolysaccharide binding protein reveals the location of a frequent mutation that impairs innate immunity.
Immunity
39
:
647
660
.
72
Lamping
N.
,
Hoess
A.
,
Yu
B.
,
Park
T. C.
,
Kirschning
C. J.
,
Pfeil
D.
,
Reuter
D.
,
Wright
S. D.
,
Herrmann
F.
,
Schumann
R. R.
.
1996
.
Effects of site-directed mutagenesis of basic residues (Arg 94, Lys 95, Lys 99) of lipopolysaccharide (LPS)-binding protein on binding and transfer of LPS and subsequent immune cell activation.
J. Immunol.
157
:
4648
4656
.
73
Heinzelmann
M.
,
Polk
H. C.
 Jr.
,
Miller
F. N.
.
1998
.
Modulation of lipopolysaccharide-induced monocyte activation by heparin-binding protein and fucoidan.
Infect. Immun.
66
:
5842
5847
.
74
Rasmussen
P. B.
,
Bjørn
S.
,
Hastrup
S.
,
Nielsen
P. F.
,
Norris
K.
,
Thim
L.
,
Wiberg
F. C.
,
Flodgaard
H.
.
1996
.
Characterization of recombinant human HBP/CAP37/azurocidin, a pleiotropic mediator of inflammation-enhancing LPS-induced cytokine release from monocytes.
FEBS Lett.
390
:
109
112
.
75
Hreggvidsdottir
H. S.
,
Ostberg
T.
,
Wähämaa
H.
,
Schierbeck
H.
,
Aveberger
A. C.
,
Klevenvall
L.
,
Palmblad
K.
,
Ottosson
L.
,
Andersson
U.
,
Harris
H. E.
.
2009
.
The alarmin HMGB1 acts in synergy with endogenous and exogenous danger signals to promote inflammation.
J. Leukoc. Biol.
86
:
655
662
.
76
Youn
J. H.
,
Oh
Y. J.
,
Kim
E. S.
,
Choi
J. E.
,
Shin
J. S.
.
2008
.
High mobility group box 1 protein binding to lipopolysaccharide facilitates transfer of lipopolysaccharide to CD14 and enhances lipopolysaccharide-mediated TNF-α production in human monocytes.
J. Immunol.
180
:
5067
5074
.
77
Bosshart
H.
,
Heinzelmann
M.
.
2002
.
Arginine-rich cationic polypeptides amplify lipopolysaccharide-induced monocyte activation.
Infect. Immun.
70
:
6904
6910
.
78
Berbée
J. F.
,
Coomans
C. P.
,
Westerterp
M.
,
Romijn
J. A.
,
Havekes
L. M.
,
Rensen
P. C.
.
2010
.
Apolipoprotein CI enhances the biological response to LPS via the CD14/TLR4 pathway by LPS-binding elements in both its N- and C-terminal helix.
J. Lipid Res.
51
:
1943
1952
.
79
Berbée
J. F.
,
van der Hoogt
C. C.
,
Kleemann
R.
,
Schippers
E. F.
,
Kitchens
R. L.
,
van Dissel
J. T.
,
Bakker-Woudenberg
I. A.
,
Havekes
L. M.
,
Rensen
P. C.
.
2006
.
Apolipoprotein CI stimulates the response to lipopolysaccharide and reduces mortality in Gram-negative sepsis.
FASEB J.
20
:
2162
2164
.
80
Heinzelmann
M.
,
Platz
A.
,
Flodgaard
H.
,
Polk
H. C.
 Jr.
,
Miller
F. N.
.
1999
.
Endocytosis of heparin-binding protein (CAP37) is essential for the enhancement of lipopolysaccharide-induced TNF-α production in human monocytes.
J. Immunol.
162
:
4240
4245
.
81
Kaiserman
D.
,
Bird
P. I.
.
2010
.
Control of granzymes by serpins.
Cell Death Differ.
17
:
586
595
.
82
Wells
M. J.
,
Sheffield
W. P.
,
Blajchman
M. A.
.
1999
.
The clearance of thrombin-antithrombin and related serpin-enzyme complexes from the circulation: role of various hepatocyte receptors.
Thromb. Haemost.
81
:
325
337
.
83
Strickland
D. K.
,
Kounnas
M. Z.
.
1997
.
Mechanisms of cellular uptake of thrombin-antithrombin II complexes role of the low-density lipoprotein receptor-related protein as a serpin-enzyme complex receptor.
Trends Cardiovasc. Med.
7
:
9
16
.
84
Lu
F.
,
Lamontagne
J.
,
Sun
A.
,
Pinkerton
M.
,
Block
T.
,
Lu
X.
.
2011
.
Role of the inflammatory protein serine protease inhibitor Kazal in preventing cytolytic granule granzyme A-mediated apoptosis.
Immunology
134
:
398
408
.
85
Spaeny-Dekking
E. H.
,
Kamp
A. M.
,
Froelich
C. J.
,
Hack
C. E.
.
2000
.
Extracellular granzyme A, complexed to proteoglycans, is protected against inactivation by protease inhibitors.
Blood
95
:
1465
1472
.
86
Sun
J.
,
Bird
C. H.
,
Sutton
V.
,
McDonald
L.
,
Coughlin
P. B.
,
De Jong
T. A.
,
Trapani
J. A.
,
Bird
P. I.
.
1996
.
A cytosolic granzyme B inhibitor related to the viral apoptotic regulator cytokine response modifier A is present in cytotoxic lymphocytes.
J. Biol. Chem.
271
:
27802
27809
.
87
Rowshani
A. T.
,
Strik
M. C.
,
Molenaar
R.
,
Yong
S. L.
,
Wolbink
A. M.
,
Bemelman
F. J.
,
Hack
C. E.
,
Ten Berge
I. J.
.
2005
.
The granzyme B inhibitor SERPINB9 (protease inhibitor 9) circulates in blood and increases on primary cytomegalovirus infection after renal transplantation.
J. Infect. Dis.
192
:
1908
1911
.
88
Kurschus
F. C.
,
Kleinschmidt
M.
,
Fellows
E.
,
Dornmair
K.
,
Rudolph
R.
,
Lilie
H.
,
Jenne
D. E.
.
2004
.
Killing of target cells by redirected granzyme B in the absence of perforin.
FEBS Lett.
562
:
87
92
.
89
Wang
L.
,
Li
Q.
,
Wu
L.
,
Liu
S.
,
Zhang
Y.
,
Yang
X.
,
Zhu
P.
,
Zhang
H.
,
Zhang
K.
,
Lou
J.
, et al
.
2013
.
Identification of SERPINB1 as a physiological inhibitor of human granzyme H.
J. Immunol.
190
:
1319
1330
.
90
de Koning
P. J.
,
Kummer
J. A.
,
de Poot
S. A.
,
Quadir
R.
,
Broekhuizen
R.
,
McGettrick
A. F.
,
Higgins
W. J.
,
Devreese
B.
,
Worrall
D. M.
,
Bovenschen
N.
.
2011
.
Intracellular serine protease inhibitor SERPINB4 inhibits granzyme M-induced cell death.
PLoS ONE
6
:
e22645
.
91
Mahrus
S.
,
Kisiel
W.
,
Craik
C. S.
.
2004
.
Granzyme M is a regulatory protease that inactivates proteinase inhibitor 9, an endogenous inhibitor of granzyme B.
J. Biol. Chem.
279
:
54275
54282
.
92
Wilharm
E.
,
Parry
M. A.
,
Friebel
R.
,
Tschesche
H.
,
Matschiner
G.
,
Sommerhoff
C. P.
,
Jenne
D. E.
.
1999
.
Generation of catalytically active granzyme K from Escherichia coli inclusion bodies and identification of efficient granzyme K inhibitors in human plasma.
J. Biol. Chem.
274
:
27331
27337
.
93
Lim
Y. P.
,
Bendelja
K.
,
Opal
S. M.
,
Siryaporn
E.
,
Hixson
D. C.
,
Palardy
J. E.
.
2003
.
Correlation between mortality and the levels of inter-alpha inhibitors in the plasma of patients with severe sepsis.
J. Infect. Dis.
188
:
919
926
.

The authors have no financial conflicts of interest.