Role Of Granzyme Research Articles

Abstract 4984: TCR-CD3 bispecifics kill tumor cells in a caspase- and gasdermin-dependent manner by inducing multiple cell death pathways

Abstract Background:ImmTAC® (immune-mobilizing monoclonal T cell receptor against cancer) molecules are soluble bi-specific TCR molecules that redirect T lymphocytes to kill tumor cells. Tebentafusp, which targets the melanocyte lineage gp100, is the first ImmTAC molecule approved for the treatment of HLA-A*02:01 adult patients with metastatic uveal melanoma.1 While the roles of Granzyme B and Perforin in the anti-tumor activity of CD3 T cell engagers are well described, the contribution of other apoptotic pathways, e.g. pyroptosis (inflammatory apoptosis), is poorly understood. For this reason, we explored apoptosis mechanisms induced by ImmTACs in vitro and their relevance to the anti-tumor activity of tebentafusp in a Phase 2 clinical trial. Methods: Cell lines derived from different malignancies, MP41 (uveal melanoma), MEL624 (cutaneous melanoma) and NCI-H1755 (non-small cell lung carcinoma) were co-cultured with PBMC and ImmTAC molecules for defined time periods, followed by cytolysis measurements (xCelligence) and protein expression analysis (western blotting). Relevant cell death pathways were targeted by genetic (CRISPR knockout) or pharmacological inhibition. Translational relevance of in vitro experiments was assessed using RNAseq of tumor biopsies at baseline (N=70) and matched on-treatment at 16 days post treatment [N=35] or at progression [N=14]) from a Phase 2 trial of tebentafusp in HLA-A*02:01+ mUM patients (NCT02570308). Clinical data cut-off: 17-Oct-2022. Results: ImmTAC-mediated tumor killing in vitro occurred in a caspase-dependent manner and accompanied a profound induction of several apoptotic, necroptotic and pyroptotic proteins, such as GZMB, Caspases 8 and 7, RIPK3, MLKL and GSDMD in several cell lines. In NCI-H1755 cells, genetic knockout of GSDMD significantly diminished ImmTAC-mediated cytolysis (92% reduction), thereby confirming the involvement of pyroptosis in ImmTAC-mediated tumor killing in these cells. In agreement, RNAseq analysis of tumor biopsies from mUM patients after 3 doses of tebentafusp revealed enhanced expression of genes involved in different cell death pathways, including Caspase 7, RIPK3, MLKL and GSDMD (fold changes of 1.2-2). In addition, patients exhibiting above-median tumor expression of GSDMD at progression had longer overall survival (Hazard ratio = 0.074 95% C.I. 0.01-0.64). Conclusions: ImmTAC redirection of T cells resulted in a caspase-dependent, pyroptotic cell death of tumor cell lines and enhanced gene expression of necroptotic and pyroptotic pathways in tumor biopsies. GSDMD, induced during ImmTAC-mediated cytolysis, also correlated with better overall survival in tebentafusp-treated patients. Induction of multiple cell death pathways, particularly pyroptosis, helps to explain the anti-tumor activity of tebentafusp.

The Role of Granzyme B Activity and Concentration, and Serpin B9 Concentration with Hepatocellular Carcinoma in Patients with Chronic Hepatitis B Virus

Backgrounds: Granzyme B (GzmB) is the main effective weapon of adaptive immune response that involved in killing of the infective as well as cancerous cells. Serum concentration and/or activity of GzmB may influence the course of chronic hepatitis B (CHB). Aims of the study: To investigate the role of serum level of GzmB and its activity as well as the effect of serpin B9 inhibitor on serum level of GzmB in patients with HCC. Patients and Methods: This is a cross-sectional study which included 85 patients with CHB. Those patients were divided into two groups: Group 1: includes 45 patients diagnosed with HCC, group 2: includes 40 Patients without HCC. Demographic data of the patients were collected from direct interview. Liver function test were measured spectrophotometrically. Granzyme B activity was estimated using spectrophotometry. Enzyme linked immunosorbent assay was used to measure Granzyme B concentration, serum level of and serpin B9. Results: Patients without HCC had significantly higher activity (median: 58 pmol, range: 34.0-130.0 pmol) and concentration (median; 31 pg/ml, range: 0.0-383.0 pg/ml) than those without HCC (median activity: 26, range: 7.0-63.0, median concentration: 18.7 pg/ml, range: 0.0-52.0 pg/ml) with highly significant differences. Conclusions: Granzyme B activity and concentration are significantly reduced in CHB patients with HCC compared with those without HCC. Thus, the reduction in GzmB activity cannot be attributed to increase Serpin B9, rather the GzmB polymorphism may be the main effector. Keywords: Chronic hepatitis B, Hepatocellular carcinoma, Granzyme B activity and concentration, Serpin B9

Granzyme B in epithelial barrier dysfunction and related skin diseases.

The predominant function of the skin is to serve as a barrier-to protect against external insults and to prevent water loss. Junctional and structural proteins in the stratum corneum, the outermost layer of the epidermis, are critical to the integrity of the epidermal barrier as it balances ongoing outward migration, differentiation, and desquamation of keratinocytes in the epidermis. As such, epidermal barrier function is highly susceptible to upsurges of proteolytic activity in the stratum corneum and epidermis. Granzyme B is a serine protease scarce in healthy tissues but present at high levels in tissues encumbered by chronic inflammation. Discovered in the 1980s, granzyme B is currently recognized for its intracellular roles in immune cell-mediated apoptosis as well as extracellular roles in inflammation, chronic injuries, tissue remodeling, as well as processing of cytokines, matrix proteins, and autoantigens. Increasing evidence has emerged in recent years supporting a role for granzyme B in promoting barrier dysfunction in the epidermis by direct cleavage of barrier proteins and eliciting immunoreactivity. Likewise, granzyme B contributes to impaired epithelial function of the airways, retina, gut, and vessels. In the present review, the role of granzyme B in cutaneous epithelial dysfunction is discussed in the context of specific conditions with an overview of underlying mechanisms as well as utility of current experimental and therapeutic inhibitors.

Elevated serum granzyme B levels are associated with disease activity and joint damage in patients with rheumatoid arthritis.

ObjectivesLittle is known about the roles of granzyme B in rheumatoid arthritis (RA). We aimed to evaluate the serum level of granzyme B in patients with RA and determine relationships with clinical features and joint destruction of RA.MethodsWe enrolled 100 patients with RA, 50 patients with osteoarthritis (OA), and 50 healthy controls (HC). Granzyme B serum concentrations were measured by ELISA; we then analyzed associations between granzyme B levels, clinical features, and joint destruction by calculating Sharp scores and disease activity as measured by Disease Activity Score-28 based on erythrocyte sedimentation rate (DAS28-ESR) in patients with RA.ResultsCompared with HC and patients with OA, serum granzyme B levels in patients with RA were remarkably elevated. Serum granzyme B levels did not differ between patients with OA and HC. Granzyme B levels correlated with ESR, rheumatoid factor, swollen joint counts, joint erosion scores, total Sharp scores, and DAS28-ESR. Moreover, patients with RA with high disease activity had higher granzyme B levels.ConclusionsSerum granzyme B levels were elevated significantly in patients with RA and correlated positively with disease activity and joint destruction. Serum granzyme B may have potential applications in laboratory evaluation of patients with RA.

The Role of Granzyme B Containing Cells in the Progression of Chronic Obstructive Pulmonary Disease.

BackgroundLung inflammation plays a vital role in the pathogenesis of chronic obstructive pulmonary disease (COPD), but the characteristics of the inflammatory process remain unclear. There is growing interest in the role of granzyme B (GzmB) because CD8+ T cells can induce apoptosis of target cells by releasing GzmB, which in turn may cause tissue injury and remodeling. However, GzmB is also expressed by regulatory cells, which are able to suppress CD8+ T cell. The role of GzmB+ cells needs to be defined in COPD.MethodsGzmB+ and CD8+ cells on alveolar wall of surgically resected lungs of microscopically classified 12 nonsmoking control, 12 panlobular emphysema (PLE) and 30 centrilobular emphysema (CLE) subjects were localized by immunohistochemical method. Positively stained cells on alveolar wall were counted and length of corresponding alveolar wall was measured. The results were expressed as mean number of positively stained cells per mm of alveolar wall in each subject.ResultsThe number of GzmB+ and CD8+ cells on alveolar wall of CLE was greater than that of control or PLE subjects (p<0.05 and p<0.001, respectively). There was a positive relationship between the number of alveolar GzmB+ cells and forced expiratory volume in 1 second (FEV1) (r=0.610, p=0.003) in CLE subjects. The number of alveolar GzmB+ cells progressively decreased with decline of FEV1.ConclusionOur finding that number of alveolar GzmB+ cells was associated with FEV1 suggests that GzmB+ cells might have protective role in the progression of lung destruction and airflow limitation in CLE, which is the predominant emphysema subtype of COPD.

Granzymes A and K differentially potentiate LPS-induced cytokine response.

Granzymes are serine proteases that, upon release from cytotoxic cells, induce apoptosis in tumor cells and virally infected cells. In addition, a role of granzymes in inflammation is emerging. Recently, we have demonstrated that extracellular granzyme K (GrK) potentiates lipopolysaccharide (LPS)-induced cytokine response from monocytes. GrK interacts with LPS, disaggregates LPS micelles, and stimulates LPS-CD14 binding and Toll-like receptor signaling. Here we show that human GrA also potentiates cytokine responses in human monocytes initiated by LPS or Gram-negative bacteria. Similar to GrK, this effect is independent of GrA catalytic activity. Unlike GrK, however, GrA does not bind to LPS, has little influence on LPS micelle disaggregation, and does not augment LPS-CD14 complex formation. We conclude that GrA and GrK differentially modulate LPS-Toll-like receptor signaling in monocytes, suggesting functional redundancy among cytotoxic lymphocyte proteases in the anti-bacterial innate immune response.

Roles of granzyme A in pathology of patients with atopic dermatitis

Roles of granzyme A in pathology of patients with atopic dermatitis

Elucidating Sources and Roles of Granzymes A and B during Bacterial Infection and Sepsis

During bacterial sepsis, proinflammatory cytokines contribute to multiorgan failure and death in a process regulated in part by cytolytic cell granzymes. When challenged with a sublethal dose of the identified mouse pathogen Brucella microti, wild-type (WT) and granzyme A (gzmA)(-/-) mice eliminate the organism from liver and spleen in 2 or 3 weeks, whereas the bacteria persist in mice lacking perforin or granzyme B as well as in mice depleted of Tc cells. In comparison, after a fatal challenge, only gzmA(-/-) mice exhibit increased survival, which correlated with reduced proinflammatory cytokines. Depletion of natural killer (NK) cells protects WT mice from sepsis without influencing bacterial clearance and the transfer of WT, but not gzmA(-/-) NK, cells into gzmA(-/-) recipients restores the susceptibility to sepsis. Therefore, infection-related pathology, but not bacterial clearance, appears to require gzmA, suggesting the protease may be a therapeutic target for the prevention of bacterial sepsis without affecting immune control of the pathogen.

Altered sputum granzyme B and granzyme B/proteinase inhibitor‐9 in patients with non‐eosinophilic asthma

The non-eosinophilic phenotype of asthma (NEA) is associated with chronic airway inflammation and airway neutrophilia. An accumulation of apoptotic airway epithelial cells, if not efficiently cleared by efferocytosis, can undergo secondary necrosis, with the potential for inflammation of surrounding tissues. Apoptosis may occur via the T cell granzyme B pathway. The role of granzyme B in NEA is not known. The aim of this study was to investigate production of granzyme B and its inhibitor proteinase inhibitor (PI)-9 by T cells from induced sputum and compare expression between eosinophilic, NEA and healthy controls. We investigated T cell intracellular granzyme B and its inhibitor, PI-9, in sputum from healthy control subjects (n = 10), and patients with NEA (n = 22) or eosinophilic asthma (EA) (n = 15) using flow cytometry. Granzyme B expression and the ratio of granzyme B to PI-9 positive cells were highest in those with NEA for both CD3+ and CD4+ T cells. The expression of granzyme B was not statistically different between patients with NEA and EA; however, the ratio of granzyme B to PI-9 positive cells for CD3+ T cells was significantly higher in those with NEA compared with EA. Induced sputum provides a non-invasive tool for investigating T cell cytotoxic mediators in the various asthma subtypes. Granzyme B expression is increased in NEA and the contribution of granzyme B to chronic inflammation requires further study.

Granzyme B cleavage of fibronectin disrupts endothelial cell adhesion, migration and capillary tube formation

Dysregulated angiogenesis contributes to the pathogenesis of chronic inflammatory diseases. Modulation of the extracellular matrix by immune-derived proteases can alter endothelial cell–matrix interactions as well as endothelial cell sprouting, migration and capillary formation. Granzyme B is a serine protease that is expressed by a variety of immune cells, and accumulates in the extracellular milieu in many chronic inflammatory disorders that are associated with dysregulated angiogenesis. Although granzyme B is known to cleave fibronectin, an essential glycoprotein in vascular morphogenesis, the role of granzyme B in modulating angiogenesis is unknown. In the present study, granzyme B cleaved both plasma fibronectin and cell-derived fibronectin, resulting in the release of multiple fibronectin fragments. Granzyme B cleavage of fibronectin resulted in a dose-dependent reduction in endothelial cell adhesion to fibronectin as well as reduced endothelial cell migration and tubular formation. These events were prevented when granzyme B activity was inhibited by a small molecule inhibitor. In summary, granzyme B-mediated cleavage of fibronectin contributes to attenuated angiogenesis through the disruption of endothelial cell — fibronectin interaction resulting in impaired endothelial cell migration and tubular formation.

Granzymes are necessary for suppressive function of regulatory T cells

Purpose Regulatory T (Treg) cells are an essential subset of CD4+ T cells that induce and maintain immunological tolerance. Preclinical animal models have demonstrated that adoptive transfer of Treg cells can prevent or cure diabetes, multiple sclerosis (EAE), inflammatory bowel disease, lupus, arthritis, and graft versus host disease. Defects in Treg cell function and number have been described in a number of different human autoimmune diseases including diabetes, multiple sclerosis, rheumatoid arthritis and juvenile idiopathic arthritis. These data suggest that manipulation of Treg cells may be a useful therapeutic intervention. Treg cells are marked by expression of the forkhead/winged helix transcription factor Foxp3, which is essential for their regulatory functions. Treg cells are well known to mediate their immunomodulatory effects through TGF-b, IL-10 and CTLA-4. Recent work has also suggested a role for the granule/exocytosis pathway in Treg cell suppressive function. Granzyme A and granzyme B (gzm A and gzm B, respectively) mRNAs are expressed at high levels in Treg cells, and Treg cells from gzm B–/– mice showed defects in suppression in vitro. To further examine the role of granzymes in the function of Treg cells in vivo, we used a T cell transfer model of colitis. In this model, naive CD45RBhigh CD4+ T cells transferred to RAG–/– mice and undergo homeostatic expansion and activation.This process results in colitis manifested clinically by weight loss and shortened survival. Previous work has shown that a subset of the transferred conventional T cells develop into Foxp3+ iTreg cells and that these iTreg cells are necessary to mitigate colitis when present together with natural Treg cells (nTreg) cells derived in the thymus. Methods CD45RBhigh T cells from mice expressing a Foxp3EGFP fusion protein and additionally deficient in gzm A or gzm B are transferred to RAG-/mice.In these mice, EGFP expression marks iTreg which have developed in situ. Mice were weighed twice weekly and euthanized when moribund or when they have lost more than 1520% of their initial weight. Mesenteric lymph nodes T cells were analyzed for expression of EGFP.

Granzyme B as a novel factor involved in cardiovascular diseases

Apoptosis plays an important role in cardiovascular diseases such as atherosclerosis, ischemic heart disease, and congestive heart failure. Previous studies have demonstrated that oxidative stress, physiological stress, and inflammatory cytokines such as tumor necrosis factor and Fas ligand are involved in apoptosis of cardiovascular system. We demonstrate that another apoptosis-related pathway, i.e. granzyme B/perforin system is involved in cardiovascular diseases. Expression of granzyme B, a member of serine protease family is increased in acute coronary syndrome, coronary artery disease with end-stage renal disease, and subacute stage of acute myocardial infarction. Although granzyme B is extensively researched in immunological disorders, the role of granzyme B/perforin system was not clear in the cardiovascular field. In addition, little is known regarding the inhibition of granzyme B system in the clinical situation. In this review we demonstrate recent findings of granzyme B in cardiovascular diseases and possible therapeutic applications of inhibiting the granzyme B/perforin system.

A Role for Granzyme M in TLR4-Driven Inflammation and Endotoxicosis

Lymphocyte perforin and serine protease granzymes are well-recognized extrinsic mediators of apoptosis. We now demonstrate that cytotoxic lymphocyte granule components profoundly augment the myeloid cell inflammatory cytokine cascade in response to TLR4 ligation. Whereas caspase-1-deficient mice were completely resistant to LPS, reduced serum cytokine production and resistance to lethal endotoxicosis were also obtained with perforin-deficient mice, indicating a role for granzymes. Consistently, a lack of granzyme M (GrzM) resulted in reduced serum IL-1alpha, IL-1beta, TNF, and IFN-gamma levels and significantly reduced susceptibility to lethal endotoxicosis. These altered responses were also observed in granzyme A-deficient but not granzyme B-deficient mice. A role for APC-NK cell cross-talk in the inflammatory cascade was highlighted, as GrzM was exclusively expressed by NK cells and resistance to LPS was also observed on a RAG-1/GrzM-double deficient background. Collectively, the data suggest that NK cell GrzM augments the inflammatory cascade downstream of LPS-TLR4 signaling, which ultimately results in lethal endotoxicosis. Most importantly, these data demonstrate that granzymes should no longer be considered solely as mediators of apoptosis, but additionally as potential key regulators of inflammation.

Role of Granzyme B in the Antitumor Effects Induced by Recipient Lymphocyte Infusion in Mixed Bone Marrow Chimeras

Role of Granzyme B in the Antitumor Effects Induced by Recipient Lymphocyte Infusion in Mixed Bone Marrow Chimeras

Research on expression of granzyme B in myocardium with experimental viral myocarditis

Objective To explore the role of granzyme B( GzmB) in infiltrated myocardial cells of mice with viral myocarditis( VMC) induced by coxsackievirus B3(CVB3) during the injury of myocardium and further to elucidate the pathogenesis of VMC. Methods Thirty-six BALB/c male mice, 5 to 6 weeks old, were randomly divided into two groups: the normal group, 10, and the model group, 26. On the eleventh day all mice were killed and the hearts were taken out for inspection whether there were pathological changes in the myocardium by macroscopy and microscopy. The expression of GzmB was detected in myocardium with immunohistochemistry and hemi-quantitive reverse transcript-polymerase chain reaction ( RT-PCR). Results GzmB positive cells in the mice myocardial cells infiltrated were detected by immunohistochemistry in the model group,and the number of GzmB positive cells is remarkable positive correlation to myocardial pathological score(r =0. 859, P =0.000) , however, no such cells were discovered in the normal murine hearts. Test of RT-PCR showed markedly high expression of GzmB mRNA in mice myocardium of the model group, however, no expression of GzmB mRNA in the normal group. Conclusion The expression of GzmB in the infiltrating myocardiocytes probably plays some role during the injury of myocardium, which mediated cytotoxicity effect is one of important mechanisms of myocardial injury in viral myocarditis. The expression quantity of GzmB in the infiltrating myocardiocytes is significant positive correlation to the severity of myocardial injury. Key words: Viral myocarditis; T lymphocytes; cytotoxic; Killer cells; Granzyme B

The possible role of granzyme B in the pathogenesis of chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is a highly prevalent inflammatory lung condition characterized by airways disease and emphysema, and the precise mechanism of pathogenesis is poorly understood. The consistent features of COPD include protease-antiprotease imbalance, inflammation and accelerated aging caused by apoptosis or senescence. One family of molecules involved in all of these processes is the granzymes, serine proteases with the best-known member being granzyme B (GzmB). The majority of GzmB is released unidirectionally towards target cells, but GzmB can also be released nonspecifically and escape into the extracellular environment. GzmB is capable of cleaving extracellular matrix (ECM) proteins in vitro, and the accumulation of GzmB in the extracellular milieu during chronic inflammation in COPD could contribute to ECM degradation and remodelling and, consequently, the emphysematous phenotype in the lung. Preliminary studies suggest that increased GzmB expression is associated with increased COPD severity, and this may represent a promising new target for drug and biomarker discovery in COPD. In this paper, we review the potential pathogenic contributions of GzmB to the pathogenesis of COPD.

Differential Expression of Granzyme B and C in Murine Cytotoxic Lymphocytes

Cytotoxic lymphocytes use the granule exocytosis pathway to kill pathogen-infected cells and tumor cells. Although many genes in this pathway have been extensively characterized (e.g., perforin, granzymes A and B), the role of granzyme C is less clear. We therefore developed a granzyme C-specific mAb and used flow cytometry to examine the expression of granzyme B and C in the lymphocyte compartments of wild-type and mutant GzmB(-/-) cre mice, which have a small deletion in the granzyme B gene. We detected granzyme B and C expression in CD4(+) and CD8(+) T cells activated with CD3/CD28 beads or MLRs. Stimulation of NK cells in vitro with IL-15 also induced expression of both granzymes. Granzyme C up-regulation was delayed relative to granzyme B in wild-type lymphocytes, whereas GzmB(-/-) cre cells expressed granzyme C earlier and more abundantly on a per-cell basis, suggesting that the deleted 350-bp region in the granzyme B gene is important for the regulation of both granzymes B and C. Quantitative RT-PCR revealed that granzyme C protein levels were regulated by mRNA abundance. In vivo, a population of wild-type CD8alphaalpha(+) intraepithelial lymphocytes constitutively expressed granzyme B and GzmB(-/-) cre intraepithelial lymphocytes likewise expressed granzyme C. Using a model of a persistent murine CMV infection, we detected delayed expression of granzyme C in NK cells from infected hosts. Taken together, these findings suggest that granzyme C is activated with persistent antigenic stimulation, providing nonredundant backup protection for the host when granzyme B fails.

Abstract 2668: Critical Role of Granzyme B in Left Ventricular Remodeling after Acute Myocardial Infarction

Background: Granzyme B is a member of the serine esterase family produced by cytotoxic T lymphocytes (CTLs), and has an important role in cellular apoptosis and extracellular matrix degradation. We hypothesized that granzyme B is involved in left ventricular (LV) remodeling after acute myocardial infarction (AMI). Objectives: To elucidate the role of granzyme B in LV remodeling after AMI. Subjects and methods: We employed 41 patients with the first AMI (mean age: 61.9±8.9 years old). We obtained peripheral blood on day 1, day 7 and day 14 after onset. Plasma levels of apoptosis-related molecules, including tumor necrosis factor α (TNFα), a soluble form of the Fas ligand (sFasL) and granzyme B were measured. We checked the activation of CTLs by flow cytometry. Patients were treated by percutaneous coronary intervention within 12 hours after onset. Successful myocardial reperfusion (TIMI flow grade 2 or 3) was accomplished in all patients. LV end-diastolic volume index (LVEDVI) was calculated on day 1 and 6 months after onset. Results: Plasma levels of TNFα, sFasL and granzyme B increased significantly after the onset of AMI (TNFα; day 1: 1.6±0.47, day 7: 3.3±0.65, day14: 4.0±1.2 pg/ml, p&lt;0.05, sFasL; day 1: 72±5.2, day 7: 86±6.7, day14: 95±7.3 pg/ml, p&lt;0.05, granzyme B; day 1: 63±18.4, day 7: 283±57.8, day14: 210.9±46.3 pg/ml, p&lt;0.001). The percentage of CD69± to CD3+ CD8+ lymphocytes was significantly increased (CD69+/CD3+ CD8+; day 1: 14.5±1.7, day 7: 15.8±1.2, day 14: 19.1±1.4%, p&lt;0.05), suggesting that CTLs were activated after onset. Univariate regression analysis showed a significant positive correlation between plasma granzyme B level on day 14 and fold-increase in LVEDVI (r=+0.45, p&lt;0.01). No significant correlation was observed between TNFα and changes in LVEDVI, or sFasL and changes in LVEDVI. Stepwise multivariate regression analysis showed that the plasma granzyme B level on day 14 is a significant explanatory variable for changes in LVEDVI (β = +0.53, p&lt;0.001). Conclusions: These results first indicate that among proapoptic molecules, granzyme B has a critical role in the progression of LV remodeling after AMI.

Abstract 3711: Increased Expression of Plasma Granzyme B in the Patients with Coronary Artery Disease Complicated with Chronic Kidney Disease

Background: Recent clinical studies have revealed that chronic kidney disease (CKD) is a significant independent risk factor for major cardiovascular events. However, the mechanisms in which CKD increases the incidence of cardiovascular diseases are still not fully understood. Hypothesis: Granzyme B is a member of serine protease family released from cytotoxic T lymphocytes and plays an important role in cellular apoptosis via an activation of caspases. We hypothesized that granzyme B is involved in formation of coronary artery lesions in patients with CKD. Objectives: To elucidate the role of granzyme B in coronary artery disease (CAD) in patients complicated with CKD. Subjects and methods: We employed 141 patients (116 men and 25 women, mean age: 64.2±9.6 years old ). Diagnosis of CAD was confirmed by selective coronary angiography. CKD was defined as sustained decrease in estimated glomerular filtration rate less than 60 mL/min/1.73m 2 over 3 months. We assigned patients into three groups; CAD without CKD (CAD group, n=46), CKD without CAD (CKD group, n=17) and CAD with CKD (CAD/CKD group, n=78). Patients with acute coronary syndrome, immunological disorder, malignancy and acute inflammatory diseases were excluded from the study. Results: Analysis of variance showed that plasma levels of high-sensitive CRP (hsCRP) and granzyme B in CAD/CKD group were significantly higher than those in the other groups (hsCRP: CAD; 2420±763, CKD; 1802±584, CAD/CKD: 7419±2171 ng/mL, p&lt;0.05, granzyme B: CAD: 9.1±2.3, CKD: 42.2±2.4, CAD/CKD: 159±51.8 pg/mL, p&lt;0.01). A significant positive correlation was observed between plasma hsCRP and granzyme B levels (r=+0.30, p&lt;0.05). A significant negative correlation was observed between eGFR and granzyme B levels (r=−0.32, p&lt;0.01). Multiple regression analysis revealed that granzyme B and hs CRP levels were independent predicting variables for the number of stenosis in major coronary arteries (F=5.35, p=0.002, beta coefficients: granzyme B=+0.24, p=0.006, hsCRP=+0.18, p=0.03). Conclusions: These results indicate that granzyme B might be a critical enzyme for formation of coronary atherosclerosis by inducing apoptosis of vascular tissues in patients with CKD.

Granzyme K: a novel mediator in acute airway inflammation

Background:Granzymes are a subfamily of serine proteases involved in the pathogenesis of many inflammatory disorders. In contrast with granzyme A and B, the role of granzyme K (GrK) in human...