transforming-growth-factor-beta and Hepatitis-B--Chronic

Hepatitis B virus (HBV) infection is a leading cause of liver damage and hepatic inflammation. Upon infection, effective antiviral responses by CD8+ T cells, CD4+ T cells, Natural killer (NK) cells, and monocytes can lead to partial or complete eradication of the viral infection. To date, many studies have shown that the production of inhibitory cytokines such as Interleukin 10 (IL-10), Transforming growth factor beta (TGF-β), along with dysfunction of the dendritic cells (DCs), and the absence of efficient innate immune responses could lead to T cell exhaustion, development of persistent infection, and inability to eradicate the viral infection from liver. Understanding the immunopathogenesis of the virus could be useful in providing further insights toward novel strategies in the eradication of HBV infection.

Topics: Antiviral Agents; Clonal Anergy; Dendritic Cells; Gene Expression Regulation; Hepatitis B Vaccines; Hepatitis B virus; Hepatitis B, Chronic; Humans; Immunity, Innate; Interleukin-10; Killer Cells, Natural; Liver; Mass Vaccination; Monocytes; T-Lymphocytes; Transforming Growth Factor beta; Viral Load

Hepatocellular carcinoma (HCC) is the leading cause of cancer death, and hepatitis B virus (HBV) infection is one of the commonest causes in Asian countries. India has the second largest pool after China for hepatitis B-infected subjects. HBV clearance is T cell dependent, and one of the reasons for T cells hyporesponsiveness is due to mass production of regulatory T cells (Tregs) through activation of Notch signalling, which suppress CD4/CD8 T cells. Tregs are important to maintain cellular homoeostasis; however, during viral infection increase of Tregs is inversely proportional to HBV DNA titres. Tregs exert their suppressive effect either via cell-to-cell contact or through release of interleukin (IL)-2, IL-10, TGF-β and IL-35. In Chronic hepatitis B virus CHBV infection, PD-1 pathway also gets activated and is involved in promoting tolerance. However, with Tregs induction, virus-specific T cell responses also get decreased. Circulatory and intratumoural Tregs promote development of HBV-specific HCC more by decreasing and impairing the effector functions of CD8 T cells. Antiviral therapies and PD-1 blockade strategy had shown the inhibition of Tregs and reduction in HBV DNA. However, inhibition of HBV-specific Tregs is major challenge for future therapies. New cytokine blockade therapies have emerged as potential therapeutic potentials.

Topics: Carcinoma, Hepatocellular; Hepatitis B virus; Hepatitis B, Chronic; Humans; India; Interleukin-1; Interleukin-10; Interleukin-2; Liver Neoplasms; Programmed Cell Death 1 Receptor; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Viral Load

Hepatitis B virus (HBV) which includes, fulminant, acute, chronic, asymptomatic, and occult HBV infection is the most prevalent virus that leads to human liver diseases. Chronic, asymptomatic, and occult infection can induce further sever diseases such as hepatocellular carcinoma (HCC) and cirrhosis of the liver. The underlying mechanisms that allow progression of the prolonged forms of the infection and subsequent HCC or cirrhosis of the liver are yet to be clarified. However, many researchers have suggested that immunological and genetic parameters may play important roles in the etiology of hepatitis B. Transforming growth factor beta (TGF-β) is an important cytokine with dual regulatory functions in the immune system and in the responses against viral infections. However, the pathways and mechanisms controlling these are not fully understood. The crucial roles of TGF-β in the development of Th17 and T regulatory lymphocytes, the main cell types involved in autoimmunity and destructive immune related diseases, have been documented and this provides insights into TGF-β function during hepatitis infection and subsequent HCC and cirrhosis of the liver. Recent findings also confirm that TGF-β directly alters hepatocyte function during hepatitis B, hence, the aim of this review is to address the current data regarding the association and status of TGF-β with hepatitis B infection and its related disorders including HCC and cirrhosis of the liver.

Topics: Carcinoma, Hepatocellular; Hepatitis B, Chronic; Humans; Liver Cirrhosis; Liver Neoplasms; Transforming Growth Factor beta

Epidemiological and clinical data point to a close association between chronic hepatitis B virus infection or chronic hepatitis C virus infection and development of hepatocellular carcinoma (HCC). HCC develops over several decades and is associated with fibrosis. This sequence suggests that persistent viral infection and chronic inflammation can synergistically induce liver fibrosis and hepatocarcinogenesis. The transforming growth factor-β (TGF-β) signaling pathway plays a pivotal role in diverse cellular processes and contributes to hepatic fibro-carcinogenesis under inflammatory microenvironments during chronic liver diseases. The biological activities of TGF-β are initiated by the binding of the ligand to TGF-β receptors, which phosphorylate Smad proteins. TGF-β type I receptor activates Smad3 to create COOH-terminally phosphorylated Smad3 (pSmad3C), while pro-inflammatory cytokine-activated kinases phosphorylates Smad3 to create the linker phosphorylated Smad3 (pSmad3L). During chronic liver disease progression, virus components, together with pro-inflammatory cytokines and somatic mutations, convert the Smad3 signal from tumor-suppressive pSmad3C to fibro-carcinogenic pSmad3L pathways, accelerating liver fibrosis and increasing the risk of HCC. The understanding of Smad3 phosphorylation profiles may provide new opportunities for effective chemoprevention and personalized therapy for patients with hepatitis virus-related HCC in the future.

Topics: Animals; Carcinoma, Hepatocellular; Cell Transformation, Viral; Hepatitis B, Chronic; Hepatitis C, Chronic; Humans; JNK Mitogen-Activated Protein Kinases; Liver; Liver Neoplasms; Phosphorylation; Receptors, Transforming Growth Factor beta; Risk Assessment; Risk Factors; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta

To discuss exhanstively: complex molecular and cellular mechanism in Hepatocellular Carcinoma (HCC); effect of chronic inflammation and cirrhosis, accompained by regenerative process, on the development of HCC; genetic instability of liver cells of regenerating nodules; the relative role of hepatitis C virus (HCV) and hepatitis B virus (HBV) in hepatocarcinogenesis; tumorigenicity of aflatoxin B1 (AFB1); gene expression profiles in HCC; liver tumors and host defense; future perspectives of HCC treatment.. We reviewed the most important studies on HCV.. HCC is an aggressive malignancy with poor prognosis and is one of the most common tumor in the world. In the majority of cases, HCC is found in conjunction with cirrhosis of the liver. Chronic inflammation and cirrhosis, accompagnied by regenerative process, function as a tumor promoter, providing a common pathway from chronic HBV or HCV infection to HCC. The direct etiologic role of HBV and HCV for HCC is obscure. Tumor progression may be brought about in HCC by mutation of the p53 tumor suppressor gene. The prevalences of p53 mutations is similar in HBV-associated and HCV-associated HCCs. Another mechanisms of host defense are the production of transforming growth factor beta1 (TGFbeta1), and the induction of cytotoxic T lymphocytes; the failure of there mechanisms permits the process of hepatocarcinogenesis. Treatment with alpha interferon of chronic hepatitis is necessary to delary or prevent the progression to liver cirrhosis and development of HCC. Various therapies, such radical operation, intra-arterial chemoembolization, percutaneous intratumoral ethanol injection, radio-frequency ablation, have been employed, but there is still non satisfactory treatment. Recent advances in recombinant and gene delivery thechnologies suggest that gene therapy may be a promising alternative to explore. Furthemore, immunotherapy may become a modality for patients with HCC. Clinical application of vaccine immunotherapy with NY-ESO-1 derived peptides in HLA-A2 positive HCC patients will be possible.

Topics: Aflatoxin B1; Animals; Carcinoma, Hepatocellular; Controlled Clinical Trials as Topic; Gene Expression; Genes, p53; Genetic Therapy; Hepatectomy; Hepatitis B, Chronic; Hepatitis C, Chronic; Humans; Immunotherapy; Immunotherapy, Active; Interferon-alpha; Liver Cirrhosis; Liver Neoplasms; Liver Regeneration; Mice; Mice, Transgenic; Multicenter Studies as Topic; Mutation; Prognosis; Prospective Studies; Randomized Controlled Trials as Topic; Retrospective Studies; T-Lymphocytes, Cytotoxic; Transforming Growth Factor beta

Chronic hepatitis B and C virus infections have been characterized by the pathophysiological features with a high incidence of progression to cirrhosis and development of hepatocellular carcinoma. The viral persistence produced by escape mutations from virus-specific cytotoxic T lymphocytes (CTL) response may lead to upregulation of delayed-type hypersensitivity immune response, which causes hepatic tissue damage through non specific macrophage activation and CTL response and promotes pathogenesis of hepatic fibrosis. In a preliminary clinical study, a novel metalloendopeptidase-F (MEP-F) has been shown to be effective in the treatment of patients with either chronic hepatitis B or C infection. Oral administration of MEP-F resulted in a significant reduction of the serum levels of HBs antigen and HCV RNA and improvement in the liver function abnormalities. However, the mechanism of action of MEP-F is not yet well understood. There are accumulating evidences showing an important role of alpha 2-macroglobulin-proteinase complexes in regulatory mechanisms of immune response and repairing within impaired and inflammatory tissues. In this article, reviewing the pharmacological and biological properties of alpha 2-macroglobulin-proteinase complexes, the mechanism of anti-viral effect of MEP-F is examined based on the clinical findings. It is indicated that alpha 2-macroglobulin-MEP-F complexes may induce macrophage/Kuppfer cell activation and proliferation through binding their receptors on the cells and activating signaling cascades, which enhance both anti-viral specific and nonspecific immune responses. alpha 2-Macroglobulin-MEP-F complexes may also augment cellular immunity and hepatic regeneration by neutralizing the immunosuppressive and fibrogenic activities of transforming growth factor-beta.

Topics: alpha-Macroglobulins; Animals; Cytokines; Hepatitis B, Chronic; Hepatitis C, Chronic; Humans; Liver; Liver Regeneration; Low Density Lipoprotein Receptor-Related Protein-1; Macrophages; Metalloendopeptidases; Receptors, Immunologic; T-Lymphocytes, Cytotoxic; Transforming Growth Factor beta

Transforming growth factor (TGF)- beta1, metalloproteinase (MMP)-1 and its tissue inhibitor (TIMP)-1 are considered predictive biomarkers of chronic hepatitis activity and fibrosis. The aim of this study was to evaluate the effect of lamivudine treatment on the plasma levels of these peptides in patients with chronic hepatitis B.. TGF-beta1, MMP-1 and TIMP-1 plasma concentrations were measured with an enzyme immunoassay in 40 patients treated with lamivudine for 48 wk. Elimination of HBV-DNA and HBV antigens was evaluated 24 wk after treatment completion.. Baseline TGF-beta1 (29.6+/-2.2 ng/mL) and TIMP-1 (1 578+/-93 ng/mL) significantly exceeded normal values (18.3+/-1.6 ng/mL and 1 102+/-67 ng/mL respectively). Lamivudine treatment resulted in a significant decrease of TGF-beta1 and TIMP-1 during treatment with an increase after 24 wk of treatment. Pretreatment MMP-1 levels (6.7+/-0.7 ng/mL) were significantly lower than normal values (11.9+/-0.9 ng/mL) and increased during treatment and follow-up. A significant correlation was noted between TGF-beta1 or TIMP-1 and aminotransferases as well as fibrosis scored in liver biopsy specimens. There were no statistically significant differences of TGF-beta1, TIMP-1 and MMP-1 between four groups at baseline, 24 and 48 wk of treatment. TGF-beta1 and TIMP-1 levels increased significantly in non-responders and normalized in responders at wk 72. MMP-1 also normalized in responders and decreased to values significantly lower than normal in non-responders.. These findings support the role of TGF-beta1, TIMP-1 and MMP-1 in the pathogenesis of chronic hepatitis B. Because of their association with hepatic injury and antiviral treatment efficacy, determination of these peptides may be useful in disease management.

Topics: Adult; Alanine Transaminase; Aspartate Aminotransferases; Female; Hepatitis B, Chronic; Humans; Lamivudine; Liver Cirrhosis; Male; Matrix Metalloproteinase 1; Middle Aged; Reverse Transcriptase Inhibitors; Tissue Inhibitor of Metalloproteinase-1; Transforming Growth Factor beta; Transforming Growth Factor beta1

This study aims to investigate the correlations between gene alterations induced in Mdr2-knockout (Mdr2-/-) models and liver fibrosis.. The overlapping genes in Mdr2-/- models were determined and included in logistic regression analysis to identify potential candidates for predicting liver fibrosis. Correlations between the expression levels of the identified candidates and hepatic stellate cells (HSCs) were addressed. Functional enrichment of the identified candidates was also evaluated via bioinformatic analysis.. Twenty-two overlapping genes in the GSE4612, GSE8642 and GSE14539 datasets were identified. Univariate and multivariate analysis indicated that ELOVL fatty acid elongase 7 (ELOVL7) was significantly associated with liver fibrosis S ≥ 2 (OR = 11.8, 95% CI = 2.0 - 69.2, p = 0.006). ELOVL7 was significantly upregulated in patients with various types of liver injury including hepatitis B virus (HBV) infection and fatty liver diseases, and in multiple liver injury models, including bile duct ligation (BDL), carbon tetrachloride (CCl4) and paracetamol injection-induced liver damage models (all p < 0.05). The ELOVL7 levels were significantly higher in HSCs than in other liver cells (all p < 0.05) and were significantly upregulated in activated HSCs compared to quiescent HSCs (all p < 0.05). In addition, ELOVL7 expression was positively associated with transforming growth factor β (TGFβ) and bone morphogenic protein 9 (BMP9) expression and negatively associated with BMP7 expression. Bioinformatic analysis of functional enrichment indicated that ELOVL7 is mainly involved in fatty acid synthesis and metabolism.. ELOVL7 could accurately predict advanced liver fibrosis. It might be involved in the activation of HSCs and the TGFβ signaling pathway.

Topics: Fatty Acid Elongases; Hepatic Stellate Cells; Hepatitis B; Hepatitis B virus; Hepatitis B, Chronic; Humans; Liver; Liver Cirrhosis; Transforming Growth Factor beta

Chronic HBV infection is always accompanied by differences in the balance between regulatory T cells (Tregs) and T-helper 17 (Th17) cells in infection phases. IL-21 plays an important role in the progression of chronic HBV infection. Thus, the aim of our study was to investigate the role of the regulatory function of IL-21 in maintaining the balance between Tregs and Th17 cells in chronic HBV infection.. Twenty-five chronic HBV-infected patients in the immune-tolerant (IT) phase and 23 chronic hepatitis B (CHB) patients were recruited in this study. Cytokines production was measured by ELISA. The mRNA expression levels were determined by qPCR. CD4. The concentration of IL-21 in the serum of CHB were significantly higher than that in the serum from IT patients, and IL-21 and IL-21R levels in the PBMCs from CHB were higher than those from IT patients. IL-21 promoted Th17 cells differentiation and function but inhibited Treg cells differentiation and function by activating STAT3 signaling pathways, upregulating RORγt expression, downregulating Foxp3 expression, by increasing IL-17and IL-22 secretion, and decreasing TGF-β secretion in chronic HBV infection. The proportion of Tregs and TGF-β concentrations in CHB was significantly lower than that in IT patients. Furthermore, the percentage of Th17 cells and the IL-17 concentration in CHB was markedly higher than that in IT patients, causing a reduction in the Tregs/Th17 ratio in CHB patients.. Our results suggest that IL-21 may contribute to inflammation in chronic HBV infection by modulating the balance between Treg and Th17 cells.

Topics: Hepatitis B virus; Hepatitis B, Chronic; Humans; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

The relationship between the serum transforming growth factor (TGF)-β level and HBsAg loss has not been clearly elaborated in patients with chronic hepatitis B (CHB).. Two cohorts of patients with CHB were studied. Cohort A: A total of 207 hepatitis B e antigen (HBeAg)-negative CHB patients who finished ≥1 year nucleos(t)ide analogue monotherapy and sequentially received PEGylated interferon treatment for less than 96 weeks were included. Cohort B: Forty HBeAg-positive patients who initially received entecavir therapy for at least 96 weeks were included. Their viral markers and serum TGF-β levels were measured at different time points during therapy.. The levels of serum TGF-β and HBsAg (0-24 W) were significantly lower in the patients who had HBsAg< 0.05 IU/mL at 48 weeks than in patients who did not in cohort A. We got the same results when we further divided the patients into subgroups according to the initial HBsAg cut-off values (1000 IU/mL, 100 IU/mL, 50 IU/mL) in cohort A. However, HBeAg seroconversion did not lead to the downregulation of TGF-β levels. The levels of serum TGF-β were significantly correlated with HBsAg quantitation in cohort A (12-24 W) but not in cohort B (0-48 W). The levels of TGF-β at week 12 could be used as an early index to predict a functional cure (AUC=0.818) as well as the levels of HBsAg itself (AUC=0.882) in HBeAg-negative chronic hepatitis B patients treated with PEGylated interferon.. The levels of serum TGF-β were significantly associated with HBsAg loss but not with HBeAg seroconversion and could be used as an early index to predict a functional cure in CHB patients treated with PEGylated interferon.

Topics: Antiviral Agents; DNA, Viral; Hepatitis B e Antigens; Hepatitis B Surface Antigens; Hepatitis B virus; Hepatitis B, Chronic; Humans; Interferon-alpha; Polyethylene Glycols; Recombinant Proteins; Transforming Growth Factor beta; Transforming Growth Factors; Treatment Outcome

Chronic liver fibrosis is an inevitable stage for the development of patients with chronic hepatitis B (CHB). However, anti-fibrotic therapies have been unsuccessful so far. The biological functions and molecular mechanisms of long non-coding RNAs (lncRNAs) in the host immune system during chronic hepatitis B virus (HBV) infection, especially in fibrosis, are still largely unknown.. The total RNA of peripheral blood mononuclear cells (PBMCs) from asymptomatic carriers (ASCs) or CHB receiving at least 8 years of anti-viral treatments was analyzed using Arraystar microarray and validated. A total of 1,042 mRNA transcripts (630 up-regulated and 412 down-regulated) were identified being differentially expressed between ASC and CHB patients. Through enrichment analysis we focused on the transforming growth factor beta (TGF-. These findings not only added knowledge to the understanding of the roles of which lncRNA-HEIM played in the activation of HSCs in CHB patients with long-term medication, but also provided a promising therapeutic target in the future treatment for liver fibrosis.

Topics: Adult; Antiviral Agents; Carrier State; Female; Hepatic Stellate Cells; Hepatitis B virus; Hepatitis B, Chronic; Humans; Leukocytes, Mononuclear; Liver Cirrhosis; Male; Middle Aged; Monocytes; RNA, Long Noncoding; RNA, Messenger; Signal Transduction; Smad4 Protein; Transforming Growth Factor beta

Long non-coding RNA-ATB (LncRNA-ATB) which is activated by transforming growth factor-β (TGF-β), is a key regulator of TGF-β signaling pathway. TGF-β plays an important role in various pathogenic processes, from inflammation and fibrosis to cirrhosis and cancer. In this study, we evaluated the plasma levels of lncRNA-ATB in patients with hepatitis B virus (HBV)-related cirrhosis and non-cirrhotic patients with chronic hepatitis B (CHB) and investigated the clinical values. Plasma samples were collected from 44 HBV-related cirrhosis patients, 45 non-cirrhotic CHB and 75 healthy controls. Briefly, after total RNA extraction and cDNA synthesis, quantitative real-time PCR (qPCR) was performed to detect plasma lncRNA-ATB levels. Results show the plasma levels of lncRNA-ATB in HBV-related cirrhosis patients were significantly higher in comparison to healthy controls (Fold change=2.60, p value=0.04). Also, we determined plasma levels of lncRNA-ATB as a specific biomarker of HBV-related cirrhosis (AUC=0.65, p value=0.03, Sensitivity 61.36%; Specificity 70.00%). In addition to, we investigated the plasma levels of lncRNA-ATB in non-cirrhotic CHB patients were significantly lower than healthy controls (Fold change= 0.33, p value=0.01). We also indicated plasma lncRNA-ATB levels were as a sensitive biomarker for diagnosis of non-cirrhotic CHB patients compared with healthy (AUC=0.66, p value=0.00, Sensitivity 71.11%; Specificity 57.78%). According to our results, circulating lncRNA-ATB has good specificity for diagnosing hepatitis B virus (HBV)-related cirrhosis and good sensitivity for diagnosis of non-cirrhotic chronic hepatitis B (CHB) patients.

Topics: Biomarkers; Hepatitis B virus; Hepatitis B, Chronic; Humans; Liver Cirrhosis; RNA, Long Noncoding; Transforming Growth Factor beta

The characteristics and functions of CD4

Topics: CD4 Antigens; Dendritic Cells; Hepatitis B, Chronic; Humans; Immune Tolerance; Interleukin-10; Interleukin-2 Receptor alpha Subunit; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

An HBsAg-HBIG therapeutic vaccine (Yeast-derived Immune Complexes, YIC) for chronic hepatitis B (CHB) patients has undergone a series of clinical trials. The HBeAg sero-conversion rate of YIC varied from 21.9% to 14% depending on the immunization protocols from 6 to 12 injections. To analyze the immunological mechanisms exerted by 6 injections of YIC, 44 CHB patients were separately immunized with YIC, alum as adjuvant control or normal saline as blank control, with add on of antiviral drug Adefovir in all groups. Kinetic increase in Th1 and Th2 cells CD4

Topics: Adenine; Adjuvants, Immunologic; Adult; Antigen-Antibody Complex; Combined Modality Therapy; Female; Hepatitis B Surface Antigens; Hepatitis B Vaccines; Hepatitis B, Chronic; Humans; Interleukin-10; Interleukin-17; Interleukin-2; Male; Organophosphonates; Th1 Cells; Th2 Cells; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Young Adult

Hepatitis B is an immune response-mediated disease. The aim of this study was to explore the differences of ratios of T-helper (Th) 2 cells to Th1 cells and cytokine levels in acute hepatitis B (AHB) patients and chronic hepatitis B virus (HBV)-infected patients in immune-tolerance and immune-active phases.. Thirty chronic HBV-infected patients in the immune-tolerant phase (IT group) and 50 chronic hepatitis B patients in the immune-active (clearance) phase (IC group), 32 AHB patients (AHB group), and 13 healthy individuals (HI group) were enrolled in the study. Th cell proportions in peripheral blood, cytokine levels in plasma, and serum levels of HBV DNA, hepatitis B surface antigen, and hepatitis B e antigen were detected.. The Th1 cell percentage and Th2/Th1 ratio in the HBV infection group (including IT, IC, and AHB groups) were significantly different from those in HI group (24.10% ± 8.66% and 1.72 ± 0.61 vs. 15.16% ± 4.34% and 2.40 ± 0.74, respectively; all P < 0.001). However, there were no differences in the Th1 cell percentages and Th2/Th1 ratios among the IT, IC, and AHB groups. In HBV infection group, the median levels of Flt3 ligand (Flt3L), interferon (IFN)-γ, and interleukin (IL)-17A were significantly lower than those in HI group (29.26 pg/ml, 33.72 pg/ml, and 12.27 pg/ml vs. 108.54 pg/ml, 66.48 pg/ml, and 35.96 pg/ml, respectively; all P < 0.05). IFN-α2, IL-10, and transforming growth factor (TGF)-β2 median levels in hepatitis group (including patients in AHB and IC groups) were significantly higher than those in IT group (40.14 pg/ml, 13.58 pg/ml, and 557.41 pg/ml vs. 16.74 pg/ml, 6.80 pg/ml, and 419.01 pg/ml, respectively; all P < 0.05), while patients in hepatitis group had significant lower Flt3L level than IT patients (30.77 vs. 59.96 pg/ml, P = 0.021). Compared with IC group, patients in AHB group had significant higher median levels of IL-10, TGF-β1, and TGF-β2 (22.77 pg/ml, 10,447.00 pg/ml, and 782.28 pg/ml vs. 8.66 pg/ml, 3755.50 pg/ml, and 482.87 pg/ml, respectively; all P < 0.05).. Compared with chronic HBV-infected patients in immune-tolerance phase, chronic HBV-infected patients in immune-active phase and AHB patients had similar Th2/Th1 ratios, significantly higher levels of IFN-α2, IL-10, and TGF-β. AHB patients had significantly higher IL-10 and TGF-β levels than chronic HBV-infected patients in immune-active phase.

Topics: Adult; Aged; Cytokines; Female; Hepatitis B; Hepatitis B virus; Hepatitis B, Chronic; Humans; Interferon-alpha; Interferon-gamma; Interleukin-10; Male; Middle Aged; Th1 Cells; Th2 Cells; Transforming Growth Factor beta; Young Adult

MicroRNAs (miRNAs) may serve as potential molecular markers to predict liver injury resulting from chronic hepatitis B (CHB). In the present study, we want to study the expression profile and clinical significance of miRNAs at different stages of CHB virus infection.. Using miRNA microarray, we investigated the global expression profiles of cellular miRNA in asymptomatic hepatitis B antigen carriers (ASCs) and CHB patients, compared with healthy controls (HCs).. We identified 79 and 203 differentially expressed miRNAs in the peripheral blood mononuclear cells of ASCs and CHB patients compared to HCs, respectively. Some of these miRNAs were common to ASCs and CHB patients, but another set of miRNAs that showed differential expression between ASCs and CHB patients was also identified. Gene ontology and pathway enrichment analysis showed that the target genes of the identified miRNAs played a role in important biological functions, such as learning or memory, cell-cell adherens junction, ion channel inhibitor activity, TGF-beta signaling pathway, and p53 signaling pathway.. We identified some significant differentially expressed miRNA in different phases of HBV infection, which might serve as biomarkers or therapeutic targets in the future.

Topics: Adult; Aged; Biomarkers; Carrier State; Female; Gene Expression Profiling; Gene Ontology; Hepatitis B Surface Antigens; Hepatitis B virus; Hepatitis B, Chronic; Humans; Leukocytes, Mononuclear; Liver; Male; MicroRNAs; Middle Aged; Transforming Growth Factor beta

Natural killer (NK) cells can induce liver fibrosis remission by killing hepatic stellate cells (HSCs) and producing interferon (IFN)-γ in a mouse model; however, their anti-fibrotic immune-characteristics and regulatory mechanisms by HSCs remain to be determined, especially in livers from HBV-infected liver cirrhosis (LC) patients. We analyzed frequency, phenotype and anti-fibrotic function of hepatic and peripheral NK subsets in 43 HBV-LC patients. We found that hepatic NK subsets from LC patients displayed a decreased frequency, activation status and anti-fibrotic activity compared with those from chronic hepatitis B patients, which were mainly mediated by increased intrahepatic tumour-growth factor (TGF)-β because blockade of TGF-β significantly reversed NK anti-fibrotic function in vitro. In vivo, hepatic NK cells were enriched in proximity to the α-smooth muscle actin (α-SMA+) area within mild fibrosis regions; while in severe fibrotic areas, they were either directly attached to or separated from the α-SMA+ region. NK cells from LC patients could enter HSCs to form emperipolesis (a cell-in-cell structure) and become apoptotic; anti-TGF-β treatment ameliorated this emperipolesis. This finding suggested a novel mechanism by which activated HSCs impair NK cells' anti-fibrosis capacity through a TGF-β-dependent emperipolesis in LC patients, providing an anti-fibrotic rational by enhancing NK cell activity.

Topics: Adult; Aged; Animals; Emperipolesis; Female; Fibrosis; Hepatic Stellate Cells; Hepatitis B, Chronic; Humans; Interferon-gamma; Killer Cells, Natural; Liver; Liver Cirrhosis; Male; Mice; Middle Aged; Transforming Growth Factor beta

CD4+CD25+ regulatory T cells (Tregs) are involved in the regulation of physiological and pathological hepatic immune responses, but the roles are not well explored in natural killer (NK) cell-mediated liver diseases. In this study, using the NK cell-mediated oversensitive liver injury model of hepatitis B virus transgenic (HBs-Tg) mice triggered by a low dose of concanavalin A, it was observed that an increased number of CD4+CD25+Foxp3+ Tregs were accumulated in the liver, along with the recovery of liver injury. Adoptive transfer of hepatic Tregs from HBs-Tg mice but not wild B6 mice could significantly attenuate the oversensitive liver injury via inhibiting liver accumulation and decreasing NK cell group 2D-mediated activation of NK cells in the recipient HBs-Tg mice. Furthermore, upregulated expression of membrane-bound TGF-β (mTGF-β) and OX40 on hepatic Tregs were demonstrated to account for inhibiting the NK cell-mediated hepatic injury in HBs-Tg mice through cell-cell contact, confirmed by antibody blockade and cell Transwell experiments in vivo and in vitro. Our findings for the first time indicated that CD4+CD25+ Tregs directly suppressed NK cell-mediated hepatocytotoxicity through mTGF-β and OX40/OX40L interaction in a cell-cell contact manner in HBV-associated liver disease.

Topics: Adoptive Transfer; Animals; Cell Membrane; Concanavalin A; Cytotoxicity, Immunologic; Disease Models, Animal; Hepatitis B virus; Hepatitis B, Chronic; Immune Tolerance; Interleukin-2 Receptor alpha Subunit; Killer Cells, Natural; Liver; Lymphocyte Activation; Male; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Mice, Transgenic; NK Cell Lectin-Like Receptor Subfamily K; OX40 Ligand; Receptors, OX40; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Tumor Necrosis Factors

Transforming growth factor (TGF)-β induces cell growth arrest in well-differentiated hepatocellular carcinoma (HCC) while hepatitis B virus X protein (HBx) minimizes the tumor suppression of TGF-β signaling in early chronic hepatitis B. However, how to reverse the oncogenic effect of HBx and sustain the tumor-suppressive action of TGF-β has yet to be investigated. The present study examined the effect of TGF-β and a c-Jun N-terminal kinase (JNK) inhibitor on cell growth in HCC cells with forced expression of HBx. It was found that HBx promoted cell growth via activation of the JNK/pSMAD3L pathway and inhibition of the transforming growth factor-beta type I receptor (TβRI)/pSMAD3C pathway. pSMAD3L/SMAD4 and pSMAD3C/SMAD4 complexes antagonized each other to regulate c-Myc expression. In the absence of HBx, TGF-β induced cell growth arrest through activation of the TβRI/pSMAD3C pathway in well-differentiated HCC cells. In the presence of HBx, TGF-β had no effect on cell growth. JNK inhibitor SP600125 significantly reversed the oncogenic action of HBx and favored TGF-β to regain the ability to inhibit the cell growth in HBx-expressing well-differentiated HCC cells. In conclusion, targeting JNK signaling favors TGF-β to block HBx-induced cell growth promotion in well-differentiated HCC cells. As an adjunct to anti-viral therapy, the combination of TGF-β and inhibition of JNK signaling is a potential therapy for HBV-infected HCC.

Topics: Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Hepatitis B virus; Hepatitis B, Chronic; Humans; JNK Mitogen-Activated Protein Kinases; Liver Neoplasms; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-myc; Signal Transduction; Smad3 Protein; Smad4 Protein; Trans-Activators; Transforming Growth Factor beta; Viral Regulatory and Accessory Proteins

To investigate the role of Th17/Treg balance in immune mechanism in severe exacerbation of hepatitis B.. The clinical data of 41 patients with chronic hepatitis B were collected, and according to the conditions during hospitalization, these patients were divided into exacerbation group (19 patients) and improvement group (22 patients). On admission, at weeks 1 and 2 of treatment, and at the end of treatment, flow cytometry was used to measure the frequencies of Th17 and Treg cells in peripheral blood, and enzyme-linked immunosorbent assay was used to determine the serum levels of interleukin-17 (IL-17), interleukin-10 (IL-10), and transforming growth factor-β (TGF-β). The dynamic changes in the frequencies of Th17 and Treg cells were compared between the two groups, and the correlation between clinical indices for hepatitis and cytokines was analyzed. The t-test was used for comparison between groups, a one-way analysis of variance was used for comparison within one group across different time points, and Pearson correlation analysis was performed.. With disease progression, the exacerbation group showed an increase in the frequency of Th17 cells and a relatively low frequency of Treg cells; compared with the improvement group, the exacerbation group had a higher frequency of Th17 cells and a lower frequency of Treg cells. Th17/Treg ratio gradually increased with exacerbation and decreased with improvement in conditions; in the exacerbation group and the improvement group, Th17/Treg ratio was positively correlated with total bilirubin and negatively correlated with prothrombin activity. In the exacerbation group and the improvement group, Th17 cells were positively correlated with IL-17, and Treg cells were positively correlated with IL-10 and TGF-β.. Th17 and Treg cells play important roles in severe exacerbation of hepatitis B, and Th17/Treg ratio may be used as an immunobiological marker for the judgment of severity during severe exacerbation of hepatitis B.

Topics: Biomarkers; Disease Progression; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Hepatitis B virus; Hepatitis B, Chronic; Humans; Interleukin-10; Interleukin-17; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

Chronic hepatitis B virus (HBV) infection is a major global health burden. Functional exhaustion and numerical reduction of HBV-specific cytotoxic T lymphocytes (CTLs) in the liver and peripheral blood limit anti-HBV CTL activity in patients with chronic HBV infection (CHB). However, the ongoing anti-HBV CD8(+) T cell responses in the lymphoid organs are largely unknown due to the infeasibility of obtaining lymphoid organs from CHB patients. Here we demonstrate that the percentage of HBV-specific CD8(+) T cells is higher in the spleen of CHB patients than that from peripheral blood and liver. Although they do respond to TCR stimulation and produce IFNγ, the cells proliferate poorly. Furthermore, miR-720 expression is upregulated in HBV-specific CD8(+) T cells. Overexpression of miR-720 in primary human CD8(+) T cells inhibits TCR stimulation-induced proliferation. We also demonstrate that TGFβ sustains miR-720 upregulation after TCR stimulation, and blood TGFβ levels are associated with the outcome of type I interferon treatment of CHB patients. Thus, therapies targeting miR-720 may help restore impaired immunity in CHB patients.

Topics: Cell Cycle Proteins; Hepatitis B, Chronic; Humans; MicroRNAs; Receptors, Antigen, T-Cell; Spleen; T-Lymphocytes, Cytotoxic; Transforming Growth Factor beta

Liver fibrosis is a severe, life-threatening clinical condition resulting from nonresolving hepatitis of different origins. IL-17A is critical in inflammation, but its relation to liver fibrosis remains elusive. We find increased IL-17A expression in fibrotic livers from HBV-infected patients undergoing partial hepatectomy because of cirrhosis-related early-stage hepatocellular carcinoma in comparison with control nonfibrotic livers from uninfected patients with hepatic hemangioma. In fibrotic livers, IL-17A immunoreactivity localizes to the inflammatory infiltrate. In experimental carbon tetrachloride-induced liver fibrosis of IL-17RA-deficient mice, we observe reduced neutrophil influx, proinflammatory cytokines, hepatocellular necrosis, inflammation, and fibrosis as compared with control C57BL/6 mice. IL-17A is produced by neutrophils and T lymphocytes expressing the Th17 lineage-specific transcription factor Retinoic acid receptor-related orphan receptor γt. Furthermore, hepatic stellate cells (HSCs) isolated from naive C57BL/6 mice respond to IL-17A with increased IL-6, α-smooth muscle actin, collagen, and TGF-β mRNA expression, suggesting an IL-17A-driven fibrotic process. Pharmacologic ERK1/2 or p38 inhibition significantly attenuated IL-17A-induced HSC activation and collagen expression. In conclusion, IL-17A(+) Retinoic acid receptor-related orphan receptor γt(+) neutrophils and T cells are recruited into the injured liver driving a chronic, fibrotic hepatitis. IL-17A-dependent HSC activation may be critical for liver fibrosis. Thus, blockade of IL-17A could potentially benefit patients with chronic hepatitis and liver fibrosis.

Topics: Actins; Adult; Animals; Carbon Tetrachloride Poisoning; Carcinoma, Hepatocellular; Chemical and Drug Induced Liver Injury; Collagen; Cytokines; Female; Gene Expression Regulation; Hemangioma; Hepatectomy; Hepatic Stellate Cells; Hepatitis B, Chronic; Hepatitis, Animal; Humans; Interleukin-17; Liver Cirrhosis; Liver Neoplasms; Male; MAP Kinase Signaling System; Mice; Mice, Knockout; Middle Aged; Neutrophils; Nuclear Receptor Subfamily 1, Group F, Member 3; Protein Kinase Inhibitors; Receptors, Interleukin-17; Recombinant Proteins; Th17 Cells; Transforming Growth Factor beta

T helper cells17 (Th17) have accurate but inconclusive roles in the pathogenesis of acute-on-chronic hepatitis B liver failure (ACHBLF). Retinoic acid-related orphan receptor γ t(RORγt) and RORα are two lineage-specific nuclear receptors directly mediating Th17 differentiation. This study was aimed to evaluate the gene expression of RORα and RORγt and their potential role in ACHBLF. Forty patients with liver failure, 30 with chronic hepatitis B (CHB) and 20 healthy controls were studied. The mRNA levels of RORα and RORγt in peripheral mononuclear cells were determined by quantitative real-time polymerase chain reaction. The frequency of peripheral Th17 cells was determined using flow cytometry. The serum levels of interleukin-6(IL-6), transforming growth factor -β (TGF-β), interleukin-17(IL-17), interleukin-23(IL-23) and interferon-γ (IFN-γ) were measured by enzyme-linked immunosorbent assay. The frequency of peripheral Th17 cells in patients with liver failure was significantly increased compared to patients with CHB and controls. The peripheral mRNA levels of RORα and RORγt in hepatitis B-associated acute-on-chronic liver failure were significantly higher than in patients with CHB and controls as were the serum levels of IL-6 and TGF-β. The serum level of IFN-γ in patients with acute-on-chronic liver failure from HBV was significantly higher than patients with CHB but lower than controls. In patients with acute-on-chronic liver failure associated with HBV, RORγt, IL-6 and IL-23 were positively correlated with the frequency of Th17 cells, while RORα, TGF-β and IFN-γ had no correlation with the latter. The mRNA level of RORγt was positively correlated with model of end-stage liver disease (MELD) score, but there was no correlation of RORα and MELD score. RORγt plays an important role in the pathogenesis of acute-on-chronic HBV-associated liver failure and might be considered to be a candidate factor consistent with the severity of disease.

Topics: Adult; Female; Hepatitis B, Chronic; Humans; Interferon-gamma; Interleukin-17; Interleukin-23; Interleukin-6; Liver Failure, Acute; Lymphocyte Count; Male; Nuclear Receptor Subfamily 1, Group F, Member 1; Nuclear Receptor Subfamily 1, Group F, Member 3; RNA, Messenger; Severity of Illness Index; Th17 Cells; Transforming Growth Factor beta

Hepatitis B virus X (HBx) protein is suspected to participate in oncogenesis during chronic hepatitis B progression. Transforming growth factor beta (TGF-beta) signaling involves both tumor suppression and oncogenesis. TGF-beta activates TGF-beta type I receptor (TbetaRI) and c-Jun N-terminal kinase (JNK), which differentially phosphorylate the mediator Smad3 to become C-terminally phosphorylated Smad3 (pSmad3C) and linker-phosphorylated Smad3 (pSmad3L). Reversible shifting of Smad3-mediated signaling between tumor suppression and oncogenesis in HBx-expressing hepatocytes indicated that TbetaRI-dependent pSmad3C transmitted a tumor-suppressive TGF-beta signal, while JNK-dependent pSmad3L promoted cell growth. We used immunostaining, immunoblotting, and in vitro kinase assay to compare pSmad3L- and pSmad3C-mediated signaling in biopsy specimens representing chronic hepatitis, cirrhosis, or hepatocellular carcinoma (HCC) from 90 patients chronically infected with hepatitis B virus (HBV) with signaling in liver specimens from HBx transgenic mice. In proportion to plasma HBV DNA levels, early chronic hepatitis B specimens showed prominence of pSmad3L in hepatocytic nuclei. HBx-activated JNK/pSmad3L/c-Myc oncogenic pathway was enhanced, while the TbetaRI/pSmad3C/p21(WAF1) tumor-suppressive pathway was impaired as human and mouse HBx-associated hepatocarcinogenesis progressed. Of 28 patients with chronic hepatitis B who showed strong oncogenic pSmad3L signaling, six developed HCC within 12 years; only one of 32 patients showing little pSmad3L developed HCC. In contrast, seven of 30 patients with little Smad3C phosphorylation developed HCC, while no patient who retained hepatocytic tumor-suppressive pSmad3C developed HCC within 12 years.. HBx shifts hepatocytic TGF-beta signaling from the tumor-suppressive pSmad3C pathway to the oncogenic pSmad3L pathway in early carcinogenic process. Hepatocytic pSmad3L and pSmad3C assessment in HBV-infected liver specimens should prove clinically useful for predicting risk of HCC.

Topics: Adult; Animals; Carcinoma, Hepatocellular; Cell Nucleus; Cell Proliferation; Cell Transformation, Neoplastic; Cyclin-Dependent Kinase Inhibitor p21; DNA-Binding Proteins; DNA, Viral; Female; Hepatitis B, Chronic; Hepatocytes; Humans; JNK Mitogen-Activated Protein Kinases; Liver Cirrhosis; Liver Neoplasms; Male; Mice; Mice, Transgenic; Middle Aged; Phosphorylation; Signal Transduction; Smad3 Protein; Trans-Activators; Transcription Factors; Transforming Growth Factor beta; Viral Regulatory and Accessory Proteins; Young Adult

To investigate whether the CD4+CD25+Foxp3+ regulatory T cells are associated with serum TGF beta 1 in patients with hepatitis B.. Patients with chronic hepatitis B (CHB), chronic asymptomatic carriers (AsC), normal subjects (NS) and the resolved from HBV infection (Resolved) were recruited in this study. Flow cytometric analysis was used to detect the frequency and phenotype of peripheral CD4+CD25+Foxp3+ T cells, and Foxp3 gene expression were examined by real time PCR. Serum TGF beta 1 levels were measured by ELISA (enzyme-linked immunosorbent assay).. Patients with CHB or AsC exhibited significantly higher frequency of CD4+CD25+Foxp3+ T cells compared to healthy controls. CD4+CD25+ T cells derived from patients with CHB and AsC expressed higher level of Foxp3-mRNA. Furthermore, the frequency of CD4+CD25+Foxp3+ regulatory T cells was correlated with serum HBV DNA copy numbers in patients with CHB and AsC. Our results indicated that the serum TGF beta was increased in CHB and AsC patients compared to control patients, and that serum TGF beta was correlated with the expression of Foxp3-mRNA and the frequency of CD4+CD25+Foxp3+ regulatory T cells in patients with CHB and AsC.. The findings have important implication in the understanding of the role and mechanism of aberrant CD4+CD25+Foxp3+ regulatory T cells in the maintenance of chronicity in hepatitis B patients.

Topics: Adolescent; Adult; Carrier State; CD4 Antigens; Female; Flow Cytometry; Forkhead Transcription Factors; Hepatitis B Surface Antigens; Hepatitis B, Chronic; Humans; Interleukin-2 Receptor alpha Subunit; Male; Phenotype; Polymerase Chain Reaction; RNA, Messenger; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Viral Load; Young Adult

Topics: Adult; Connective Tissue Growth Factor; Female; Hepatitis B, Chronic; Humans; Immunohistochemistry; Liver; Male; Middle Aged; Transforming Growth Factor beta; Young Adult

Chronic activity of hepatitis B is thought to involve aberrant immune tolerance of unknown mechanism. In this study, we examined the role of CD4(+)CD25(+)Foxp3(+) regulatory T cells in disease activity and viral clearance in hepatitis B. Patients with chronic active hepatitis B (CAH) and asymptomatic HBV carriers (AsC) exhibited a significantly high frequency of CD4(+)CD25(+)Foxp3(+) T cells as opposed to that of controls and resolved HBV infection. These CD4(+)CD25(+) T cells expressed an elevated level of Foxp3 and displayed increased inhibitory activity towards both CD4(+)CD25(-) and CD8(+) effector cells. They were found to accumulate in liver biopsy tissue of CAH patients as opposed to controls. The frequency of CD4(+)CD25(+)Foxp3(+) T cells correlated positively with hepatitis B envelope (HBe) antigen status and serum HBV DNA copy numbers and had a converse relationship with HBe antibody status in patients with CAH and AsC. It was evident that in these patients, the increased frequency of CD4(+)CD25(+)Foxp3(+) T cells was associated with serum levels of transforming growth factor-beta known to promote peripheral conversion of CD4(+)CD25(-) T cells to CD4(+)CD25(+)Foxp3(+) regulatory T cells. The findings provide new insights into the role of CD4(+)CD25(+)Foxp3(+) regulatory T cells in chronic activity and viral clearance in chronic hepatitis B.

Topics: Adult; CD4 Antigens; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Forkhead Transcription Factors; Hepatitis B e Antigens; Hepatitis B, Chronic; Humans; Immunohistochemistry; Interleukin-2 Receptor alpha Subunit; Male; Reverse Transcriptase Polymerase Chain Reaction; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

This study attempted to explore the value of combining serum hepatic fibrosis-related markers and ultrasound parameters together on diagnosis of hepatic fibrosis.. Six serum markers and 8 ultrasound parameters were measured from 100 patients with chronic hepatitis B or cirrhosis. The results of the serum hepatic fibrosis-related markers and ultrasound in disease group were analyzed and compared with the findings of hepatic pathology.. By filtrating,the group of platelet derived growth factor-BB (PDGF-BB) plus hyaluronic acid (HA) plus echo characteristics of liver parenchyma (LPEC) plus length of spleen (SL) had the highest Se and Spe, which were 90.7% and 85.4% respectively.. The advantageous combination of serum markers and ultrasound parameters can significantly improve Se and Spe, which is superior to any single serum index or ultrasound parameter. And it was a better non-invasive method for diagnosing hepatic fibrosis.

Topics: Adolescent; Adult; Aged; Alanine Transaminase; Becaplermin; Biomarkers; Collagen Type III; Female; Hepatitis B, Chronic; Humans; Hyaluronic Acid; Liver Cirrhosis; Male; Middle Aged; Platelet-Derived Growth Factor; Proto-Oncogene Proteins c-sis; Reproducibility of Results; Sensitivity and Specificity; Tissue Inhibitor of Metalloproteinase-1; Transforming Growth Factor beta; Ultrasonography, Doppler, Color; Young Adult

To investigate the effect of the serum of patients with chronic hepatitis B (CHB) on apoptosis of renal tubular epithelial cells in vitro and to study the role of hepatitis B virus (HBV) and transforming growth factor-beta (1) (TGF-beta (1)) in the pathogenesis of hepatitis B virus associated glomerulonephritis (HBV-GN).. The levels of serum TGF-beta(1) were measured by specific enzyme linked immunosorbent assay (ELISA) and HBV DNA was tested by polymerase chain reaction (PCR) in 44 patients with CHB ,and 20 healthy persons as the control. The normal human kidney proximal tubular cell (HK-2) was cultured together with the sera of healthy persons, CHB patients with HBV-DNA negative (20 cases) and HBV-DNA positive (24 cases) for up to 72 h. Apoptosis and Fas expression of the HK-2 were detected by flow cytometer.. The apoptosis rate and Fas expression of HK-2 cells were significantly higher in HBV DNA positive serum group 19.01%+/-5.85% and 17.58%+/-8.35%, HBV DNA negative serum group 8.12%+/-2.80% and 6.96%+/-2.76% than those in control group 4.25%+/-0.65% and 2.33%+/-1.09%, respectively (P<0.01). The apoptosis rate and Fas expression of HK-2 in HBV DNA positive serum group was significantly higher than those in HBV DNA negative serum (P<0.01). Apoptosis rate of HK-2 cells in HBV DNA positive serum group was positively correlated with the level of HBV-DNA (r = 0.657). The level of serum TGF-beta (1) in CHB group was 163.05+/-91.35 microg/L, significantly higher as compared with 81.40+/-40.75 microg/L in the control group (P<0.01).. The serum of patients with chronic hepatitis B promotes apoptotic damage in human renal tubular cells by triggering a pathway of Fas up-regulation. HBV and TGF-beta (1) may play important roles in the mechanism of hepatitis B virus associated glomerulonephritis.

Topics: Apoptosis; Cell Line; Culture Media, Conditioned; DNA, Viral; Enzyme-Linked Immunosorbent Assay; fas Receptor; Flow Cytometry; Glomerulonephritis; Hepatitis B virus; Hepatitis B, Chronic; Humans; Kidney Tubules; Polymerase Chain Reaction; Receptors, Tumor Necrosis Factor; Transforming Growth Factor beta; Transforming Growth Factor beta1; Up-Regulation

In this study, we determined the frequencies of the genotypes associated with the polymorphism of the cytokines genes, and investigated their association with the risk of hepatocellular carcinoma (HCC) in hepatitis B virus (HBV) carriers.. Genetic polymorphism in the cytokines TNF-alpha, IFN-gamma, TGF-beta1, IL-6, and IL-10 were studied in 236 Japanese patients with HBV infection. The genetic polymorphisms of these cytokines were analyzed by polymerase chain reaction-sequence-specific primer (SSP).. There was no statistically significant difference in the genetic polymorphisms of TNF-alpha, IFN-gamma, and IL-10 genes between HBV carriers with HCC and those without HCC. However, the TGF-beta1+29 (codon 10) C/C genotype was lower in HBV carriers with HCC than in those without HCC (HCC 14.6% vs non-HCC 31.9%). The association of HCC was significantly lower in HBV carriers with C/C genotype than in those with T/C or T/T genotype in position +29 of the TGF-beta1 gene.. Our findings suggest that the genetic polymorphism in codon 10 of the TGF-beta1 gene may play a role in HCC development in patients with chronic HBV infection.

Topics: Adult; Aged; Carcinoma, Hepatocellular; Codon; Female; Genotype; Hepatitis B, Chronic; Humans; Interferon-gamma; Interleukin-10; Interleukin-6; Japan; Liver Neoplasms; Male; Middle Aged; Polymorphism, Genetic; Promoter Regions, Genetic; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

To explore the effects of transforming growth factor beta1 (TGFbeta1) gene polymorphism on liver cirrhosis induced by HBV infection and on plasma concentration of TGFbeta1.. Peripheral blood samples were collected from 134 patients with HBV-induced liver cirrhosis, which were further classified into three groups (A, B and C groups) according to Child-Pugh classification and 92 healthy blood donors. The polymorphisms at position -988, -800, -509 and codon10, codon25, and codon263 of TGFbeta1 gene were determined by PCR-ARMS- and Lightcycler respectively, combined with sequence analysis. The concentrations of TGFbeta1 and collagen type IV were measured by ELISA, and the plasma concentrations of hyaluronan and N-terminal type III procollagen peptide were measured by RIA. ARLEQUIN Ver 2.0 software was used to analyze the linkage disequilibrium between -509C > T and codon10T > C polymorphisms.. No polymorphism was found at positions -800, -988, codon25 and codon263. No significant difference for -509C > T polymorphism between the diseased and control groups was found, however, the frequencies of genotype TT and allele T at codon10 in the diseased group were 0.313 and 0.537 respectively, both significantly higher than those of the control group (0.916 and 0.440 respectively, both P < 0.05). In Child-Pugh C grade group the frequency of C allele at position -509 was 0.631, significantly higher than that of T allele (0.325), but there was no significant difference for codon10T > C polymorphism. The TGFbeta1 plasma concentration did not show any difference between the genotypes -509CC and TT in the control group, however, the TGFbeta1 plasma concentration was statistically lower in the those with TT genotype than in those with CC genotype in the diseased group. Linkage disequilibrium was found between -509C > T and codon10T > C polymorphisms and the major haplotypes were C-T and T-C, of which C-T frequency was significantly higher in the patients than in the control group. The plasma concentration of TGFbeta1 was significantly higher in the populations of haplotype C-T than in those of other haplotypes.. No relationship is found between -509C > T polymorphism of TGFbeta1 and liver cirrhosis in Chinese populations, but the presence of C allele at position -509 may play an important role in the progression of liver cirrhosis and may influence plasma concentration of TGFbeta1 in liver cirrhosis patients. Codon10T > C polymorphism may play a significant role in development of liver cirrhosis, but has no correlation with plasma concentration of TGFbeta1 and progression of liver cirrhosis. The -509C > T and codon10T > C polymorphisms are in tight linkage disequilibrium, and the major haplotypes are C-T and T-C. The haplotype C-T influences plasma concentration of TGFbeta1 and is closely related to liver cirrhosis.

Topics: Adult; Disease Progression; Female; Hepatitis B, Chronic; Humans; Liver Cirrhosis; Male; Middle Aged; Polymorphism, Genetic; Transforming Growth Factor beta; Transforming Growth Factor beta1

To observe the histological changes in the fibrotic and inflammatory tissues in response to interferon alpha treatment in patients with chronic viral hepatitis B.. Sixteen patients with chronic viral hepatitis B in S3-S4 stages established by pathological examination were treated with interferon alpha for 6-9 months, and the degree of liver fibrosis and inflammation were examined 3 times during the treatment. The expression of Fas, transforming growth factor beta1 (TGFbeta1) and HBcAg in the liver tissues were detected by immunohistochemistry, and DNA fragmentation was examined by TUNEL assay. The levels of the serum markers for liver fibrosis and liver function were also measured.. Patients with liver fibrosis in S3-S4 stages had high pathological expression of Fas and TGFbeta1 with severe DNA damage in the liver tissues. After 3 months of interferon therapy, the expression of Fas and TGFbeta1 were lowered (P<0.05), and further treatment till 3-9 months resulted in gradual decrease in the degree of hepatic fibrosis and cell apoptosis (P<0.05), with improved serum liver fibrosis indices and liver function.. Interferon alpha may alleviate liver fibrosis and suppress cell apoptosis in patients in S3-S4 stages after a 6- to 9-month continuous treatment.

Topics: Adult; Apoptosis; fas Receptor; Female; Hepatitis B, Chronic; Humans; Interferon-alpha; Liver; Liver Cirrhosis; Male; Middle Aged; Transforming Growth Factor beta

Monitoring of fibrosis process with the use of histopathologic studies on liver's bioptates is limited, so it is attempted to find reliable, obtained with less invasive methods, sensitive and reflecting fibrosis dynamics markers of this process. The aim of the study was simultaneously to assess liver's expression as well as circadian and mean daily TGF-betal concentration in serum of patients with chronic hepatitis type B in comparison to control group. Studies were performed on 50 patients (9 women, 41 men) aged 45.9 +/- 2.3 years with chronic hepatitis type B. Control group consisted of 20 patients (mean age 38.6 +/- 3.7 years), in which so called minimal changes without fibrosis were observed in histophatologic bioptate of liver. Blood for studies was collected every 4 houres during the day. Serum concentration of TGF-betal was assessed with the use of ELISA method, and expression of mRNA TGF-betal in liver with QRT-PCR method. No significant difference between circadian as well as mean daily serum TGF-betal concentration beetwen control group and the group with chronic hepatitis type B was shown. Increased expression of mRNA, TGF-betal in bioptate of liver of patients with chronic hepatitis type B in comparison to control group was noted. In "minimal changes" control group presence of significant positive correlation between expression of mRNA TGF-beta1 in liver and concentration of this cytokine in serum was shown, in the group of patients with chronic hepatitis B this connection was not noted.

Topics: Circadian Rhythm; Female; Hepatitis B, Chronic; Humans; Liver; Male; Middle Aged; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1

To measure the plasma levels of transforming growth factor beta1 (TGFbeta1), the protein expression of alpha-SMA in hepatic stellate cells and urokinase plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1), and study on the relationships between the plasma levels of TGFbeta1, the protein expression and the serum hyaluronic acid (HA) in patients with different grades of hepatic fibrosis.. Thirty seven cases with hepatic fibrosis of different grades were classified according to HE and VG staining categories from 0 to 4, in which there were 8 cases in grade 1, 9 cases in grade 2, 7 cases in grade 3, 13 cases in grade 4. The plasma levels of TGFbeta1 and the serum levels of HA were detected by ELISA. The protein expressions of a-SMA, uPA and PAI-1 in fibrotic liver tissue were observed by immunohistochemistry.. With the progression of hepatic fibrosis, the plasma levels of TGFbeta1 and the protein expression of a-SMA, uPA and PAI-1 in fibrotic liver tissue were increased. In grade 3 and 4, the plasma levels of TGFbeta and the protein expression of a-SMA and PAI-1 in fibrotic liver tissue were significantly increased, but the protein expression of uPA in cirrhosis liver tissue did not increased.. TGFbeta1, a-SMA, uPA and PAI-1 play an important role in the progression of hepatic fibrosis. Inhibiting the early activation of latent TGFbeta1 or increasing uPA and inhibiting PAI-1 over express may contribute to matrix degradation and retard the progression of hepatic fibrosis.

Topics: Actins; Adult; Aged; Female; Hepatitis B, Chronic; Humans; Liver Cirrhosis; Male; Middle Aged; Plasminogen Activator Inhibitor 1; Transforming Growth Factor beta; Transforming Growth Factor beta1; Urokinase-Type Plasminogen Activator

Fibrosis is the process accompanying majority of chronic diseases of liver, independent of etiological factor and leading to cirrhosis and hepatic failure. Monitoring fibrosis process by liver's biopsy is limited, so many attempts are undertaken to assess concentrations of definite proteins in blood, which could be easily accessible marker of intrahepatic process. It seems, that among others, determinations of blood concentration of aminoterminal propeptide of procollagen III--index of collagen's III synthesis and TGF-beta 1--cytokine of antiproliferative action and inhibiting hepatocytes' growth, yet inducing fibroblasts' growth and stimulating fibrosis process brings out such a possibility. The aim of the study was simultaneous determination of TGF-beta 1 and PIIINP concentration in blood of patients with chronic hepatitis B and C before interferone's therapy in comparison to healthy controls, assessment of the parameters in dependence on stage of liver fibrosis and determination of correlation between TGF-beta 1 and PIIINP. Studies were performed in 40 patients with chronic hepatitis B (CAH B) and 35 patients with chronic hepatitis C (CAH C). Significantly increased serum concentrations of TGF-beta 1 as PIIINP in both groups of patients (CAH B and CAH C; grading 2-3, staging 1-2) in comparison with control group was noted. Significant positive correlation of TGF-beta 1 and PIIINP serum concentrations in both groups of patients was observed. There was not significant changes in PIIINP serum levels in patients with hepatitis B and C in dependence on stage of liver fibrosis (staging 1 vs staging 2) but TGF-beta 1 serum levels was significantly increased in CAH B and C patients with higher stage of liver fibrosis process. On the base of obtained results, it seems that changes in TGF-beta 1 concentrations in blood reflect "grading" and "staging" and can be a marker of intensification of intrahepatic fibrosis process whereas PIIINP levels in blood have rather the relation with "grading".

Topics: Adult; Antigens, Viral; Antiviral Agents; Female; Hepatitis B, Chronic; Hepatitis C, Chronic; Humans; Interferons; Male; Peptide Fragments; Procollagen; Transforming Growth Factor beta; Transforming Growth Factor beta1

The aim of the study was to evaluate the serum TGF-beta 1, IL-12 and IL-5 concentration in children with chronic hepatitis (ChH) B. The study included 62 children with histopathologically diagnosed chh B. The stage of fibrosis and inflammation grade were assessed according to Batts and Ludwig and Ishak et al. The control group consisted of 9 children without clinical signs of infectious and chronic diseases. Serum TGF-beta 1 concentration was significantly elevated in patients with chronic hepatitis B (p = 0.0077) as compared to controls; there were no significant differences in serum concentrations of IL-12 and IL-5 between the examined groups of children. There was also no correlation between serum concentration of the studied cytokines and the degree of fibrosis, inflammation, activity of GPT, GOT, ALP, GGTP and concentrations of bilirubin, proteins or immunoglobulins (G, A, M).

Topics: Adolescent; Child; Child, Preschool; Female; Hepatitis B, Chronic; Humans; Interleukin-12; Interleukin-5; Male; Transforming Growth Factor beta; Transforming Growth Factor beta1

Noninvasive diagnosis of hepatic fibrosis has become the focus because of the limited biopsy, especially in the surveillance of treatment and in screening hepatic fibrosis. Recently, regulatory elements involved in liver fibrosis, such as platelet derived growth factor-BB (PDGF-BB), transforming growth factor-beta1 (TGF-beta1), matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), have been studied extensively. To determine whether these factors or enzymes could be used as the indices for the diagnosis of hepatic fibrosis, we investigated them by means of receiver operating characteristic (ROC) curve.. Serum samples from sixty patients with chronic viral hepatitis B and twenty healthy blood donors were assayed to determine the level of PDGF-BB, TGF-beta1, MMP-1, and TIMP-1 with ELISA, and HA, PCIII, C-IV, and LN level with RIA. The message RNA (mRNA) expression of TIMP-1 and MMP-1 in peripheral blood mononuclear cells (PBMCs) was detected by RT-PCR and Northern blot hybridization. Liver biopsy was performed in all patients. The biopsy samples were histopathologically examined. The trial was double-blind controlled.. The serum level of PDGF-BB, TIMP-1, the ratio of TIMP-1 and MMP-1 (TIMP-1/MMP-1), mRNA expression of TIMP-1 (TIMP-1mRNA), and the ratio of TIMP-1mRNA and MMP-1mRNA (TIMP-1mRNA/MMP-1mRNA) in patients was significantly higher than those in the healthy blood donors (t=2.514-11.435, P=0.000-0.016). The serum level of PDGF-BB, TIMP-1, TIMP-1/MMP-1, and TIMP-1mRNA was positively correlated with fibrosis stage and inflammation grade (r=0.239-0.565, P=0.000-0.033), while the serum level of MMP-1 was negatively correlated with fibrosis stage and inflammation grade, and TIMP-1mRNA/MMP-1mRNA was positively correlated with inflammation grade. Through the analysis by ROC curve, serum PDGF-BB was the most valuable marker, and its sensitivity was the highest among the nine indices. The markers with the highest specificity were TIMP-1mRNA and TIMP-1mRNA/MMP-1mRNA in PBMCs. The area under the curve (AUC) of PDGF-BB, TIMP-1mRNA, TIMP-1mRNA/MMP-1mRNA, TIMP-1/MMP-1, HA, PCIII, TIMP-1, C-IV, and LN was 0.985, 0.876, 0.792, 0.748, 0.728, 0.727, 0.726, 0.583, and 0.463, respectively. The sensitivity and the specificity in the parallel test was 99.0% and 95.0% when serum PDGF-BB, TIMP-1mRNA and TIMP-1mRNA/MMP-1mRNA was detected simultaneously.. Serum level of PDGF-BB, TIMP-1mRNA, TIMP-1mRNA/MMP-1mRNA in PBMCs, and serum level of TIMP-1 and TIMP-1/MMP-1 can be used as the indices for the diagnosis of hepatic fibrosis, but the former three are more useful. The combination of serum PDGF-BB, TIMP-1mRNA and TIMP-1mRNA/MMP-1mRNA in PBMCs is even more efficient in screening liver fibrosis.

Topics: Becaplermin; Biomarkers; Hepatitis B, Chronic; Humans; Leukocytes, Mononuclear; Liver Cirrhosis; Matrix Metalloproteinase 1; Platelet-Derived Growth Factor; Predictive Value of Tests; Proto-Oncogene Proteins c-sis; RNA, Messenger; Sensitivity and Specificity; Severity of Illness Index; Tissue Inhibitor of Metalloproteinase-1; Transforming Growth Factor beta; Transforming Growth Factor beta1

Topics: Adult; Female; Hepatitis B, Chronic; Humans; Liver Cirrhosis; Liver Function Tests; Male; Middle Aged; Transforming Growth Factor beta; Transforming Growth Factor beta1

To investigate the compositions of Th1/Th2/Th3 cells in chronic hepatitis B virus (HBV)-infected individuals by determining the expression of interleukin-4 (IL-4), inetrferon-gamma (IFN-gamma), and transform growth factor-beta (TGF-beta) in single CD4(+) T cells isolated from peripheral blood mononuclear cells (PBMCs) and the role of polarized Th cell populations in chronic HBV-infection was discussed.. PBMCs from chronically infected HBV individuals were isolated, stimulated by PMA/Ionomycin/Monensin, and IL-4, IFN-gamma and TGF-beta production by CD4(+) T cells was determined by using fluorescence activated cell sorter (FACS) analysis.. The percentage of IFN-gamma-producing T cells, IL-4-producing T cells and TGF-beta-producing T cells ranged from 2.3% - 18.6%, 1.1% - 8.7% and 0.7% - 7.1% respectively in CD4(+) T cells from non-infected individuals. Most of CD4(+) T cells from PBMCs in chronically infected HBV individuals were Th0 cells. The proportion of Th1 cells increased significantly with hepatic inflammatory activity, and in the active period of chronic hepatitis B infection were higher than those in the non-active period (P < 0.05). Th2 cell percentage in CD4(+) T cells from HBV-infected individuals did not differ significantly (P > 0.05), but were higher than that from controls (P < 0.05). Th3 cell percentage in CD4(+) T cells from asymptomatic carrier (AsC) group was higher than that in the chronic hepatitis B (CHB) and control groups (P < 0.05).. Th1 phenotype cytokines were positively correlated with hepatic inflammatory activity in chronic hepatitis B and Th2 cells may be associated with the persistence of HBV infection. Th3 cells cooperating with Th2 cells can negatively regulate immune responses and may be associated with the immune tolerant state of chronic HBV infection.

Topics: CD4-Positive T-Lymphocytes; Hepatitis B, Chronic; Humans; Interferon-gamma; Interleukin-4; T-Lymphocytes, Helper-Inducer; Th1 Cells; Th2 Cells; Transforming Growth Factor beta

To evaluate the anti-fibrotic effect of interferon alpha and to study the relationship between the anti-fibrotic effect and the efficacy of anti-virus in patients with chronic hepatitis B treated with interferon alpha.. Thirty-six patients with chronic hepatitis B were treated with interferon alpha. Before, during and after the treatment, the levels of hyaluronic acid (HA), type III procollagen (PC-III), type IV collagen(IV-C), laminin (LN) and transforming growth factor beta 1 (TGF-beta 1) were measured as the markers of hepatic fibrosis. Needle biopsy sections of the liver taken before and at the end of IFN treatment were assessed and compared according to the modified histological activity index (HAI) and hepatic fibrosis (HF) scoring system.. In IFN group, the levels of HA, PC-III, IV-C, LN, and TGF-beta 1 after treatment were all significantly lower than those before treatment (P<0.01). The levels of HA, PC-III, and TGF-beta 1 after treatment were significantly lower in IFN group than in control one. HAI scores decreased from 9.3+/-3.2 to 6.2 +/-2.1 and HF scores decreased from 7.5+/-2.2 to 5.1+/-1.8 after treatment.. The improvement of hepatic fibrosis closely related to the efficacy of anti-virus. Interferon alpha therapy is effective for hepatic fibrosis of chronic hepatitis B.

Topics: Adolescent; Adult; Collagen Type IV; Hepatitis B, Chronic; Humans; Hyaluronic Acid; Interferon-alpha; Laminin; Liver Cirrhosis; Middle Aged; Procollagen; Transforming Growth Factor beta

To investigate the composition of Th1/Th2/Th3 cells in chronic HBV-infected individuals by determining the expression of IL-4/IFN-gamma/TGF-beta in single CD4+ T cell isolated from the peripheral blood mononuclear cells (PBMCs), and analyze the role of Th cells polarized population in chronic HBV-infection.. PBMCs from chronic HBV infected individuals were separated routinely, stimulated by PMA/Ionomycin/Monensin, and the production of IL-4/IFN-gamma/TGF-beta by CD4+ T cells in PBMCs was determined by FACS analysis.. The percentage of IFN-gamma-producing T cells, IL-4-producing T cells and TGF-beta producing T cells ranged from 2.3%-18.6%, 1.1%-8.7% and 0.7%-7.1%, respectively, in CD4+ cells from non-infected individuals. The majority of CD4+ T cells from PBMCs in chronic HBV-infected individuals were Th0 cells. The proportion of Th1 cells increased significantly with the hepatic inflammation activity, and the proportion in active period of chronic hepatitis B was higher than that in nonactive period (P < 0.05). The percentage of Th2 cells in CD4+ T cells from HBV-infected individuals did not differ significantly (P > 0.05), but it was higher than controls (P < 0.05). The percentage of Th3 cells in CD4+ T cells from AsC group was higher than that in CHB group and control (P < 0.05).. Th1 phenotype cytokine are correlated with hepatic inflammation activity of chronic hepatitis B. Th2 cells may be associated with the persistence of HBV infection. Th3 cells cooperating with Th2 cells exert negative immunoregulating action and may be associated with the immune tolerance state of chronic HBV infection.

Topics: Adolescent; Adult; CD4-Positive T-Lymphocytes; Female; Flow Cytometry; Hepatitis B, Chronic; Humans; Interferon-gamma; Interleukin-4; Male; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

To observe the relation between serum transforming growth factor beta 1(TGF beta 1) and level of PCIII, LN, HA, proliferation of hepatic fibrotic tissue.. Serum TGF beta 1 was detected by ELISA in 58 patients with chronic hepatitis B(CHB) and 18 patients with liver cirrhosis(LC). 20 healthy persons served as normal control(NC). Liver puncture was performed in 29 patients with CHB. Diagnosis of pathological histology and quantitative analysis of collagenous and reticular fibers were made.. (1)Serum TGF beta 1 levels in CHB and LC were notably higher than in NC (P < 0.01) and increased successively in mild, moderate and severe degrees of CHB and LC(P < 0.01/0.05). (2)Serum TGF beta 1 was correlated with serum levels of PCIII, LN, HA(P < 0.01/0.05)positively. The quantity of hepatic collagenous and reticular fibers went up in mild, moderate and severe degrees of CHB(P < 0.01/0.05). Meanwhile serum TGF beta 1 rose at the same degree.. Serum TGF beta 1 levels increase in CHB and LC. With hepatic pathological change worsening, serum TGF beta 1 and the quantity of hepatic collagenous and reticular fibers increased to the same degree. This suggests that TGF beta 1 is a crucial factor in accelerating liver fibrosis and plays a decisive role in liver fibrosis.

Topics: Adult; Aged; Female; Hepatitis B, Chronic; Humans; Liver; Liver Cirrhosis; Male; Middle Aged; Transforming Growth Factor beta

33 specimens of liver biopsy from patients with different types of viral hepatitis were detected for transforming growth factor-beta 3(TGF-beta 3) by immunohistochemistry. The results showed that TGF-beta 3 mainly distributed in the sinusal walls cells in sinus portal, areas fibrous, septum, hepatocytes in pseudolobes and necrotic areas of severe hepatitis. The expressing intensity of TGF-beta 3, was stronger in the cirrhosis, chronic severe hepatitis than that in the chronic and acute hepatitis. As the expression of TGF-beta 3 increased, the level of serum bilirubin ascended and plasmozyme activity was prolonged. It may be concluded that the expression of TGF-beta 3 is correlated with severe hepatitis and the progress of the liver cirrhosis.

Topics: Adolescent; Adult; Female; Hepatitis B, Chronic; Hepatitis, Viral, Human; Humans; Liver; Liver Cirrhosis; Male; Middle Aged; Prognosis; Transforming Growth Factor beta; Transforming Growth Factor beta3

Transforming growth factor (TGF)-beta 1 is an important cytokine involved in the pathobiology of tissue fibrosis through its stimulation of the production of, and inhibition of the degradation of, extracellular matrix proteins. We examined the clinical usefulness of plasma TGF-beta 1 concentration as a marker of fibrogenesis in patients with chronic viral hepatitis. Thirty-five patients, 11 with minimal chronic hepatitis, 14 with mild chronic hepatitis and 10 with moderate chronic hepatitis and 20 healthy subjects were studied. Transforming growth factor-beta 1 concentrations in platelet-poor plasma were measured with a TGF-beta 1 enzyme-linked immunosorbent assay system kit after acid-ethanol extraction. Plasma TGF-beta 1 levels were significantly elevated in patients with mild and moderate chronic hepatitis, but not in those with minimal chronic hepatitis, compared with the levels in the controls. Plasma TGF-beta 1 levels were increased in parallel with the histological degree of necroinflammation and of liver fibrosis. Plasma TGF-beta 1 levels were positively correlated with blood levels of procollagen type III N-peptide, and 7S fragment and central triple-helix of type IV collagen. These results suggest that plasma TGF-beta 1 level is a useful marker in assessing the situation of liver active fibrogenesis in patients with chronic viral hepatitis.

Topics: Biomarkers; Case-Control Studies; Collagen; Enzyme-Linked Immunosorbent Assay; Female; Hepatitis B, Chronic; Hepatitis C, Chronic; Humans; Liver; Liver Cirrhosis; Male; Middle Aged; Mucins; Peptide Fragments; Procollagen; Radioimmunoassay; Transforming Growth Factor beta