31 Our results identify CB2 receptors as a novel regulator of Kupffer-cell polarization. Indeed, in vivo and in vitro experiments demonstrate PF 2341066 that genetic deletion of CB2 receptors is associated with a marked hepatic induction of the M1 signature in response to chronic alcohol feeding and a parallel loss of the M2 alternative response. These findings, therefore, suggest that endogenous

CB2 receptors are responsible for M2 response to alcohol feeding. Interestingly, the CB2 agonist, JWH-133, blunts the induction of the M1 classical signature without affecting M2 response to alcohol. Whether the lack of enhancement of M2 markers in animals treated with the CB2 agonist may be the result of partial agonist properties of the compound or to constitutive activity of CB2 receptors remains to be determined.36, 37 Nevertheless, these data demonstrate that, during chronic alcohol exposure, CB2 receptors shift the M1/M2 balance toward a predominant alternative M2 response. Besides their anti-inflammatory properties ZD1839 solubility dmso on Kupffer cells, CB2 receptors also prevent the development of

alcohol-induced fatty liver. Recent studies have demonstrated that cross-talk between Kupffer cells and hepatocytes is determinant in the control of hepatic steatosis. In rodents exposed to an alcohol diet or a high-fat diet, depletion of Kupffer cells blunts the development of fatty liver.9, 38-40 Furthermore, cocultures of M1-polarized Kupffer cells with hepatocytes promote lipid accumulation into parenchymal cells.5, 6, 38, 39 In keeping with these data, we show that CM obtained

from JWH-133- and LPS-stimulated macrophages reduces lipid accumulation in hepatocytes, compared to CM prepared from macrophages exposed to LPS alone. These data indicate that Kupffer-cell CB2 receptors decrease hepatocyte steatosis after inhibition of M1 polarization. Of note, recent studies have shown that IL-1β and TNF-α, two proinflammatory Kupffer-cell–derived cytokines, promote steatosis.38-40 We show that liver medchemexpress expression of IL-1β and TNF-α decreases in alcohol-fed mice concurrently treated with JWH-133 and increases in CB2-deficient counterparts. A similar pattern of regulation was also found in our in vitro experiments, therefore suggesting that the reduction in Kupffer-cell production of IL-1β and TNF-α may contribute to the protective effects of CB2 receptors on hepatocyte lipid accumulation. HO-1 is the rate-limiting enzyme in the catabolism of heme into biliverdin, free iron, and carbon monoxide. HO-1 is a stress-inducible protein with potent-protective effects against hepatocyte damage,41 liver inflammation,31, 33 and fibrogenesis.42, 43 Recent studies have shown that up-regulating HO-1 in Kupffer cells by means of overexpression or by pharmacological activators prevents alcohol-induced release of inflammatory mediators by Kupffer cells.31, 41 However, characterization of HO-1 inducers in Kupffer cells remains poorly documented.

Nude mice inoculated with HepG2-G2 cells developed tumors significantly larger than those developed by control mice (Fig. 8A). A considerable increase in tumor metastasis was observed in draining-tumor lymph nodes of HepG2-G2 mice with respect to controls (Fig. 8B), but no significant metastatic foci were observed in liver or lungs (data not shown). These results suggest a correlation between Gal-1 expression, tumor growth, and metastasis of

HCC. Accumulating evidence indicates that Gal-1 expression is up-regulated in hepatocarcinoma cells, yet the precise role of this lectin in liver pathophysiology is still uncertain. In the present study, we detected endogenous Gal-1 expression in HepG2 cells. Cells overexpressing Gal-1 exhibited a cytoplasmic localization of this lectin, Buparlisib manufacturer which was found

to be released to the extracellular compartment. Strikingly, if nontransfected cells were cultured continuously in the presence of rGal-1, it was detected intracellularly even after 48 hours of incubation, suggesting internalization of this protein. In fact, it has been demonstrated that Gal-1 is internalized by Jurkat leukemic T cells through receptor-mediated transport in a carbohydrate-dependent manner.22 Hence, Gal-1 secreted from HCC cells might exert its biological functions either by engaging cell surface receptors and transmitting signals inside the FK228 cell or through receptor-mediated internalization and endocytosis. However, because intracellular functions have been described for Gal-1,23 a cell surface receptor-independent mechanism responsible for Gal-1 functions medchemexpress cannot be excluded. Depending on the target

cell type and its relative concentrations within local microenvironments, Gal-1 can potentiate or inhibit cell–ECM and cell–cell interactions.5 Here, we show that both soluble and immobilized rGal-1 can promote HepG2 cell adhesion in a dose-dependent manner. Inhibition of rGal-1 effects by TDG or lactose strongly indicates a glycan-dependent mechanism in mediating Gal-1 cell adhesive properties. In fact, cell adhesion to laminin, a basement membrane glycoprotein covered by polylactosamine-enriched glycoconjugates, was also increased following exposure to soluble rGal-1. Concentrations of Gal-1 used ranged between 3.5 and 14 μM, which are in agreement with those reported for human A375 melanoma24 and ovary carcinoma cells.25 Furthermore, lower concentrations were also effective in promoting cell adhesiveness when immobilized rGal-1 was also effective when tested as an ECM protein. Moreover, the enhanced cell adhesion observed in Gal-1–transfected HepG2 cells indicates that HCC cell adhesion might be related to Gal-1 expression levels. Gal-1 can bind and form lattices with different members of the integrin family to control different biological processes, including cell adhesion, migration, proliferation, and survival.

A single “organ of interest” , the liver, has rapidly reached a status in which an entire field of study is committed to

its long-term health and longevity. Pediatric hepatologists are major components of clinical practices, education and training programs, and investigative initiatives to advance human health and to improve clinical care and outcomes. Research programs are providing insight into the combined role of genetic predisposition and environmental factors in the expression of a variety of liver diseases. This offers the opportunity to prevent or modify phenotypic expression of C59 wnt order diseases by addressing a potential chronic liver disease during early life. For example, a major current research goal is to define the pathogenesis of biliary atresia and to develop effective preventative strategies. In the interim it is important to emphasize strategies selleck kinase inhibitor for early recognition to allow for optimal intervention. Despite

the progress, our field is still quite underdeveloped. The cited advances and interventions have clearly improved outcomes for children with liver disease. Along the way we learned a great deal about hepatobiliary physiology, developmental biology, and the role of genetic variants in determining the risk of liver disease and in predicting the response to therapy. Much more needs to be done. We must focus on ensuring the continued development of our field by training the workforce of the future. In addition,

definitive, cost-effective treatment strategies must be developed such that liver transplantation may not be needed in the treatment of certain diseases, MCE such as metabolic liver disease. In this regard, it will be important for funding agencies and foundations to continue to support research and to foster innovation and collaboration in Pediatric Hepatology. The success of the well-established multicenter ChiLDREN Network serves to emphasize that point. Societies such as the AASLD and NASPGHAN must also continue to recognize the important role that pediatric hepatologists play in their mission and foster career development in the field. The emerging number of pediatric patients with nonalcoholic fatty liver disease suggests that a focus on prevention and recognition of obesity is clearly needed. This combined with efforts to prevent liver disease in early life, thoughtful medical management, precise decision-making, and conscientious, creative, and courageous use of nontransplant options can both save livers and save lives. I express my gratitude, admiration, and appreciation to all those who have made our field viable and vibrant. I especially want to recognize the commitment and collaboration of the many parents and patients who have dedicated their time to clinical studies which have clearly advanced our field. I also want to thank Mitchell Cohen and Frank DiPaola for their critical review of this article.

3 mEq/L [SD, 4.5] to 136.5 mEq/L [SD, 4.4], P = 0.0002), while the serum sodium concentration was significantly decreased from baseline in the placebo group (135.7 mEq/L [SD, 4.1]

to 135.0 mEq/L [SD, 4.3], P = 0.0060) (Table 2). Tolvaptan significantly improved ascites-related clinical symptoms, bloated feeling (P = 0.0090), malaise (P = 0.0074), sensation Cobimetinib of pressure in the decubitus position (P = 0.0017) and breathing difficulty (P = 0.0233) compared with placebo (Table 3). Forty-eight patients showed adverse events in the placebo group (60.0%) and 60 in the tolvaptan group (73.2%). Adverse events observed in the tolvaptan group during the trial period at 3% or greater frequency were thirst, constipation, renal impairment, diarrhea,

pollakiuria, pyrexia, hepatic encephalopathy, vomiting, insomnia, stomatitis find more and pruritus (Table 4). Ten patients showed serious adverse events in the placebo group (12.5%) and seven in the tolvaptan group (8.5%). Serious adverse events observed in the tolvaptan group were disseminated intravascular coagulation, liver cirrhosis, portal vein thrombosis, omphalitis, bile duct cancer, liver malignant neoplasm, hepatic encephalopathy, renal impairment, respiratory failure and hemoperitoneum (Table 5). No marked abnormalities were clinically observed in clinical laboratory tests, vital signs and 12-lead electrocardiograms. IN LIVER CIRRHOSIS patients with ascites, reduction in bodyweight from baseline was significantly greater in the tolvaptan group than in the placebo group. Reductions in abdominal circumference (parameter for assessing ascites retention) and ascites volume were greater in the tolvaptan group than in the placebo group. In addition, improvement rates of ascites-related clinical

symptoms and lower limb edema (a symptom associated with hepatic edema) were higher in the tolvaptan group than in the placebo group. Decrease in bodyweight is considered to reflect improvement of ascites and lower limb edema.[15] In this trial, ascites was considered to be improved by tolvaptan, because abdominal circumference and ascites volume both decreased, and ascites-related clinical symptoms improved. As worsening of ascites, lower limb edema and ascites-related clinical symptoms results in deterioration of QOL in patients with hepatic 上海皓元医药股份有限公司 edema,[3] tolvaptan may also improve QOL. Liver cirrhosis patients are reported to have low serum sodium concentration.[16] In this trial, mean serum sodium concentration in patients was also around the lower limit of the normal range at the start of treatment. Kim et al. reported that, in liver cirrhosis patients registered on the waiting list for liver transplantation, the hazard ratio of death increased 1.05-fold (95% CI, 1.03 to 1.08) per 1 mEq/L decrease in serum sodium concentration when serum sodium concentration was between 125 and 140 mEq/L.[17] In this trial, mean serum sodium concentration increased in the tolvaptan group and decreased in the placebo group.

3A-C). Accordingly, chimeric mice with NOX-deficient endogenous liver cells but WT BM-derived cells showed a significant reduction of αSMA MAPK Inhibitor Library research buy expression, as demonstrated by immunohistochemistry and western blotting (Fig. 3D,E). Moreover, mRNA expression for αSMA and collagen α1(I) confirmed the reduced expression of fibrogenic markers in chimeric mice with NOX-deficient HSCs (Fig. 3F). These results suggest that NOX-mediated generation of ROS in endogenous liver cells, including HSCs, is more important than in BM-derived cells, including KCs, for the development of fibrosis following cholestatic liver injury. NOX generates ROS

in many cell types. To investigate the levels of peroxidation in the NOX-chimeric livers, mice subjected to BMT Panobinostat concentration were analyzed for peroxidation by immunohistochemistry for hydroxynonenal adducts. As expected, NOX-deficient mice showed reduced peroxidation in comparison to NOX-sufficient mice. Interestingly, chimeric mice with NOX-deficient

HSCs showed a reduced level of peroxidation, confirming the importance of oxidative stress produced by NOX in HSCs during the process of liver fibrosis. (Fig. 4A). Peroxidation was also measured in whole liver samples by thiobarbituric acid reactive substances (TBARS) assays. Peroxidation in chimeric livers with NOX-deficient HSCs had a greater reduction in lipid peroxidation than the chimeric livers with NOX-deficient KCs. In fact, the level of peroxidation produced

by these chimeric mice was similar to the peroxidation in complete p47phox KO mice (Fig. 4B). To better differentiate the ROS activity in the different cell types in the liver, we performed double immunofluorescence for 4-HNE and αSMA in the chimeric mice (Fig. 4C,D). The experiment showed a colocalization of ROS production (4-HNE stain) and HSCs in chimeric mice with p47phox KO BM (p47phox KO BM WT mice) subjected to BDL (Fig. 4C), whereas HSCs express little ROS in the chimeric mice with p47phox KO endogenous liver cells (WT BM p47phox 上海皓元医药股份有限公司 KO mice) subjected to BDL (Fig. 4D), suggesting that NOX is a major contributor in HSCs. To investigate the role of NOX in a mouse model of nonalcoholic steatohepatitis (NASH) ultimately leading to fibrosis, NOX-deficient (p47phox KO) mice and WT controls were fed an MCD diet for 10 weeks. Although both WT and KO mice fed the MCD diet lost weight, the liver weight–body weight fraction revealed an increase in steatosis of the liver of all MCD-treated mice (Fig. 5A). In addition, the serum aminotransferase levels were significantly higher both in WT and KO mice fed the MCD diet than the MCS diet (Fig.

Using

a two-dimensional differential in-gel electrophoresis (DIGE) approach, different samples prelabeled with mass-matched and charge-matched fluorescent cyanine dyes (Cy2, Cy3, and Cy5) can be coseparated in the same two-dimensional gel. An internal standard is used in every gel to negate the problem of intergel variation.20, 21 In this study, a two-dimensional DIGE proteomic approach was employed to compare the proteome of Huh-7 cells overexpressing either miR-17-5p or mock controls to identify clusters of proteins (and pathways) directly or indirectly regulated by miR-17-5p. We identified a cohort of proteins indirectly regulated by miR-17-5p during the stress response, signal transduction, and metabolism. A representative molecule that was differentially phosphorylated, heat shock protein 27, was further confirmed by western

selleckchem blotting. The specific regulatory mechanisms and the effects of miR-17-5p-induced up-regulation Selleckchem GDC 0449 of phosphorylated heat shock protein 27 in HCC cells were also analyzed. DIGE, differential in-gel electrophoresis; HCC, hepatocellular carcinoma; HSP27, heat shock protein 27; MAPK, mitogen-activated protein kinase; miRNA, microRNA; MMP-2, matrix metalloproteinase 2; siRNA, small interfering RNA. For a description of the patients and methods used in this study, see the Supporting Information. The miR-17-5p up-regulation model was created using Huh-7 cells stably expressing pEZX-17-5p (clone 1 and 2). The miR-17-5p levels of clone 1 and 2 cells, pEZX-MR01-Huh-7 cells, and untreated Huh-7 cells were determined using real-time polymerase chain reaction. We observed significant up-regulation of miR-17-5p in clone 1 and 2 cells compared with mock control or untreated cells (Fig. 1A). To determine whether miR-17-5p

is associated with tumorigenesis or metastasis, clone 1 and pEZX-17-5p-Huh-7 cells were implanted through a subcutaneous route into the flanks of nude mice or orthotopically into the livers of nude mice (Fig. 1D). Up-regulation of miR-17-5p significantly facilitated overall tumor growth as assessed by measurements of tumor volume (Fig. 1B,C). The orthotopic tumor model yielded a similar tumor growth trend as seen in the original injection site. Clone 1 cells showed extensive hepatic invasion with a larger tumor volume than controls (Fig. 1E,F). medchemexpress After DIGE, the Cy2, Cy3, and Cy5 channels of each gel were individually imaged, and the images were analyzed using Decyder 5.0 software. On the basis of the average intensity ratios of protein spots, a total 30 protein spots were found to be dynamically changed in clone 1 cells (Fig. 2A), including 18 significantly up-regulated protein spots (ratioclone 1/pEZX-MR01 ≥ 1.3; P ≤ 0.05) and 12 significantly down-regulated protein spots (ratioclone 1/pEZX-MR01 ≤ 0.7; P ≤ 0.05). To identify the differentially expressed proteins in clone 1 cells, 28 protein spots with a threshold greater than 1.

Using

a two-dimensional differential in-gel electrophoresis (DIGE) approach, different samples prelabeled with mass-matched and charge-matched fluorescent cyanine dyes (Cy2, Cy3, and Cy5) can be coseparated in the same two-dimensional gel. An internal standard is used in every gel to negate the problem of intergel variation.20, 21 In this study, a two-dimensional DIGE proteomic approach was employed to compare the proteome of Huh-7 cells overexpressing either miR-17-5p or mock controls to identify clusters of proteins (and pathways) directly or indirectly regulated by miR-17-5p. We identified a cohort of proteins indirectly regulated by miR-17-5p during the stress response, signal transduction, and metabolism. A representative molecule that was differentially phosphorylated, heat shock protein 27, was further confirmed by western

click here blotting. The specific regulatory mechanisms and the effects of miR-17-5p-induced up-regulation Nivolumab ic50 of phosphorylated heat shock protein 27 in HCC cells were also analyzed. DIGE, differential in-gel electrophoresis; HCC, hepatocellular carcinoma; HSP27, heat shock protein 27; MAPK, mitogen-activated protein kinase; miRNA, microRNA; MMP-2, matrix metalloproteinase 2; siRNA, small interfering RNA. For a description of the patients and methods used in this study, see the Supporting Information. The miR-17-5p up-regulation model was created using Huh-7 cells stably expressing pEZX-17-5p (clone 1 and 2). The miR-17-5p levels of clone 1 and 2 cells, pEZX-MR01-Huh-7 cells, and untreated Huh-7 cells were determined using real-time polymerase chain reaction. We observed significant up-regulation of miR-17-5p in clone 1 and 2 cells compared with mock control or untreated cells (Fig. 1A). To determine whether miR-17-5p

is associated with tumorigenesis or metastasis, clone 1 and pEZX-17-5p-Huh-7 cells were implanted through a subcutaneous route into the flanks of nude mice or orthotopically into the livers of nude mice (Fig. 1D). Up-regulation of miR-17-5p significantly facilitated overall tumor growth as assessed by measurements of tumor volume (Fig. 1B,C). The orthotopic tumor model yielded a similar tumor growth trend as seen in the original injection site. Clone 1 cells showed extensive hepatic invasion with a larger tumor volume than controls (Fig. 1E,F). MCE After DIGE, the Cy2, Cy3, and Cy5 channels of each gel were individually imaged, and the images were analyzed using Decyder 5.0 software. On the basis of the average intensity ratios of protein spots, a total 30 protein spots were found to be dynamically changed in clone 1 cells (Fig. 2A), including 18 significantly up-regulated protein spots (ratioclone 1/pEZX-MR01 ≥ 1.3; P ≤ 0.05) and 12 significantly down-regulated protein spots (ratioclone 1/pEZX-MR01 ≤ 0.7; P ≤ 0.05). To identify the differentially expressed proteins in clone 1 cells, 28 protein spots with a threshold greater than 1.

Such unexpected results may lead to publication of provocative although counterintuitive conclusions with unpredictable consequences on clinical practice and decision-making. It is critical to the clinician that the conclusions of the Rodin study be placed

in perspective so that wise treatment and regulatory decisions can be made. This study primarily set out to prove (or otherwise) that there was no reduced inhibitor risk with plasma-derived products when compared to rFVIII products. In addition, Z-VAD-FMK the study was designed to determine whether product switching would increase the risk of inhibitor development in PUPs. These important objectives were convincingly achieved and will certainly influence the standard of care for PUPs. On the other hand, the Rodin study also suggested that a second-generation rFVIII concentrate may increase inhibitor risk and this conclusion has promulgated a loud danger signal. How this finding will be interpreted by government agencies, patient consumers, and physician prescribers may adversely affect patient, treater and health care authorities’ acceptance of such products. Provoked

PD0325901 price by the results of the Rodin study, the European Medicines Agency has recently initiated a review of the safety of the second-generation rFVIII used in the trial and intends to determine whether the marketing authorization of the product should be ‘maintained, varied, suspended, or withdrawn across the EU’ [12]. A similar fate also could potentially befall the third-generation rFVIII used in Rodin, related to the higher than anticipated PUP inhibitor incidence in the EPIC study. This commentary 上海皓元 offers an opportunity for open discussion of the results of the Rodin trial, the appropriate biostatistical approach to study design for future clinical research efforts in this field, and the relative value of a prospective/retrospective observational study vs. prospective, randomized controlled trials. CMK has received research funding from Baxter, Bayer, Grifols, Octapharma, NovoNordisk and Pfizer.

He has also served on advisory boards for Baxter, Bayer, Biogen, CSL, Grifols, Octapharma, NovoNordisk, and has consulted for all mentioned companies. He is not on any speakers bureaus but has received honoraria from all mentioned companies for providing educational programmes and participating in CME generating symposia. AI has received research funding from Baxter, Bayer, Pfizer and NovoNordisk. He has also served on advisory boards for Baxter, Bayer, Pfizer and NovoNordisk and has consulted for Bayer and NovoNordisk. He received honoraria from all mentioned companies for providing educational programmes and for participating in CME generating symposia. “
“Inhibitor development against von Willebrand factor, factor VIII or factor IX is one of the most severe complications of treating patients with von Willebrand’s disease (VWD), haemophilia A or haemophilia B respectively.

Such unexpected results may lead to publication of provocative although counterintuitive conclusions with unpredictable consequences on clinical practice and decision-making. It is critical to the clinician that the conclusions of the Rodin study be placed

in perspective so that wise treatment and regulatory decisions can be made. This study primarily set out to prove (or otherwise) that there was no reduced inhibitor risk with plasma-derived products when compared to rFVIII products. In addition, BKM120 ic50 the study was designed to determine whether product switching would increase the risk of inhibitor development in PUPs. These important objectives were convincingly achieved and will certainly influence the standard of care for PUPs. On the other hand, the Rodin study also suggested that a second-generation rFVIII concentrate may increase inhibitor risk and this conclusion has promulgated a loud danger signal. How this finding will be interpreted by government agencies, patient consumers, and physician prescribers may adversely affect patient, treater and health care authorities’ acceptance of such products. Provoked

selleck by the results of the Rodin study, the European Medicines Agency has recently initiated a review of the safety of the second-generation rFVIII used in the trial and intends to determine whether the marketing authorization of the product should be ‘maintained, varied, suspended, or withdrawn across the EU’ [12]. A similar fate also could potentially befall the third-generation rFVIII used in Rodin, related to the higher than anticipated PUP inhibitor incidence in the EPIC study. This commentary MCE offers an opportunity for open discussion of the results of the Rodin trial, the appropriate biostatistical approach to study design for future clinical research efforts in this field, and the relative value of a prospective/retrospective observational study vs. prospective, randomized controlled trials. CMK has received research funding from Baxter, Bayer, Grifols, Octapharma, NovoNordisk and Pfizer.

He has also served on advisory boards for Baxter, Bayer, Biogen, CSL, Grifols, Octapharma, NovoNordisk, and has consulted for all mentioned companies. He is not on any speakers bureaus but has received honoraria from all mentioned companies for providing educational programmes and participating in CME generating symposia. AI has received research funding from Baxter, Bayer, Pfizer and NovoNordisk. He has also served on advisory boards for Baxter, Bayer, Pfizer and NovoNordisk and has consulted for Bayer and NovoNordisk. He received honoraria from all mentioned companies for providing educational programmes and for participating in CME generating symposia. “
“Inhibitor development against von Willebrand factor, factor VIII or factor IX is one of the most severe complications of treating patients with von Willebrand’s disease (VWD), haemophilia A or haemophilia B respectively.

24, 25 Nevertheless, further studies should focus on the inclusion of the primary inflamed tissue, not only of the biliary tract but also of other affected tissues given the known tendency of IgG4-RD to either simultaneously or consecutively cause symptoms in multiple different organ systems. A gold standard for the detection of IAC is currently lacking, and diagnosis relies on the application of HISORt criteria. However, in clinical practice the discrimination between IAC, PSC, and pancreatico biliary malignancy ATR inhibitor can often only be made by the careful weighing of all clinical data, including serological tests, imaging, histology, and even response to short-term

corticosteroid treatment in selected patients. The presence

of substantial numbers of IgG4+ cells is not fully diagnostic, as IgG4+ cells can be seen in inflammatory infiltrates of other origin. These diagnostic dilemmas are underlined by the fact that several of the patients with an established PSC or pancreatico-biliary malignancy included in this study did have IgG4+ cells infiltrating the tissue, albeit in low numbers, or elevated serum IgG4 levels. To assure the validity of our findings in this study, we only included PSC and cancer patients with an unchallenged diagnosis. However, in clinical practice, it may be difficult to discriminate between, for example, a PSC patient who presents at age 52 years with elevated serum liver tests and a serum IgG4 concentration medchemexpress of 3.60 g/L but without colitis or other supporting signs, and a younger IAC patient who has the same IgG4 concentration but has no other manifestations. It is tempting to speculate Copanlisib whether these clonally expanded and class-switched B cells and plasma cells might have the ability to recognize ‘self’. Initial screens in patients with autoimmune pancreatitis have not been able to yield conclusive epitopes.26 Even though we did not find any homology

between the CDR3 sequences of the IgG4+ clones, it is possible that they harbor reactivity against the same epitope. Currently, it is impossible to predict the antigenic specificity of the B-cell receptor by its amino acid sequence. A more comprehensive set-up using high-throughput and quantitative peptide screens together with next-generation sequencing–based BCR repertoire analysis might provide more insight. In this procedure, it might be necessary to screen also for glycosylated peptides, since most organs commonly affected by IgG4-RD produce large amounts of mucins and other heavily glycosylated proteins. The specific overlap of IgG4+ clones between tissue and blood indicates that these screens can be performed on the more easily accessible peripheral blood instead of the inflamed tissue. The identification of IgG4+ clones and their antigens will help to better understand the pathogenesis of IgG4-RD, ultimately providing more targeted therapies or even cure.