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Guido Gerken

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MD , Professor of medicine , expert in hepatology , viral and autoimmune hepatitis , liver cell cancer , cirrhosis , liber. Transplantation , liver function , immunosuppressive therapy , comorbidities .

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Kusum Kharbanda

University of Nebraska Medical Center

University of Southampton

Markus Biburger

Friedrich-Alexander-Universität Erlangen-Nürnberg

Dr. Albrecht Wendel

Adri van Oudenaren

Abdur Rahman Ansari

Herold Metselaar

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Papers by Guido Gerken

Interleukin-10 expression is autoregulated at the transcriptional level in human and murine kupffer cells

Hepatology, 1998

Interleukin 10 (IL-10) is known to downregulate immune responses. The regulation of IL-10 gene ex... more Interleukin 10 (IL-10) is known to downregulate immune responses. The regulation of IL-10 gene expression therefore determines the outcome of local immune reactions. We investigated time course and downregulation of IL-10 production in primary Kupffer's cells (KC), which are known to secrete IL-10 in response to endotoxin challenge. Human and murine KC were isolated by centrifugal elutriation and investigated for IL-10 gene expression by a two-step amplification procedure (reverse transcriptasepolymerase chain reaction [PCR] followed by T7-polymerase chain reaction). We show that IL-10 messenger ribonucleic acid (mRNA) showed a G450 fold increase in KC 2 hours after endotoxin challenge. IL-10 protein release from KC strictly depended on de novo protein synthesis. Endotoxin mediated increase in IL-10 gene expression was downregulated by exogenous (G350-fold reduction of IL-10 mRNA level), as well as endogenous IL-10 protein, showing a negative autoregulatory feedback loop. IL-10 receptor expression was found to be constitutive and functional in KC. Early expression of IL-10 in KC may be of functional relevance to the outcome of immune and inflammatory reactions in the liver sinusoid. The negative autoregulation of IL-10 expression may represent a mechanism to regain a state of functional responsiveness in the microenvironment towards new proinflammatory stimuli. In conclusion, autoregulatory downregulation of IL-10 expression in KC may account for important regulatory steps of local immune response in the liver sinusoid. (HEPATOLOGY 1998;27:93-99.

IL-10 down-regulates T cell activation by antigen-presenting liver sinusoidal endothelial cells through decreased antigen uptake via the mannose receptor and lowered surface expression of accessory molecules

Clinical and Experimental Immunology, Dec 1, 1998

Our study demonstrates that antigen-presenting liver sinusoidal endothelial cells (LSEC) induce p... more Our study demonstrates that antigen-presenting liver sinusoidal endothelial cells (LSEC) induce production of interferon-gamma (IFN-g) from cloned Th1 CD4 þ T cells. We show that LSEC used the mannose receptor for antigen uptake, which further strengthened the role of LSEC as antigenpresenting cell (APC) population in the liver. The ability of LSEC to activate cloned CD4 þ T cells antigen-specifically was down-regulated by exogenous prostaglandin E 2 (PGE 2) and by IL-10. We identify two separate mechanisms by which IL-10 down-regulated T cell activation through LSEC. IL-10 decreased the constitutive surface expression of MHC class II as well as of the accessory molecules CD80 and CD86 on LSEC. Furthermore, IL-10 diminished mannose receptor activity in LSEC. Decreased antigen uptake via the mannose receptor and decreased expression of accessory molecules may explain the down-regulation of T cell activation through IL-10. Importantly, the expression of low numbers of antigen on MHC II in the absence of accessory signals on LSEC may lead to induction of anergy in T cells. Because PGE 2 and IL-10 are released from LSEC or Kupffer cells (KC) in response to those concentrations of endotoxin found physiologically in portal venous blood, it is possible that the continuous presence of these mediators and their negative effect on the local APC may explain the inability of the liver to induce T cell activation and to clear chronic infections. Our results support the notion that antigen presentation by LSEC in the hepatic microenvironment contributes to the observed inability to mount an effective cell-mediated immune response in the liver.

HIV und Schwangerschaft: Risiken der Übertragung und therapeutische Möglichkeiten

Gynakologisch-geburtshilfliche Rundschau, 1996

ABSTRACT

Viral and host factors in the prediction of response to interferon-alpha therapy in chronic hepatitis C after long-term follow-up

Journal of Viral Hepatitis, Nov 1, 1998

Chronic hepatitis caused by hepatitis C virus (HCV) infection is a common health problem affectin... more Chronic hepatitis caused by hepatitis C virus (HCV) infection is a common health problem affecting probably 3% of the world's population. Established treatment of chronic hepatitis C consists mainly of the use of interferon-α (IFN-α), which leads to a sustained treatment response in only 15-20% of patients. IFN-α therapy is frequently accompanied by many undesirable effects, e.g. headache, myalgia, thrombocytopenia or psychosis, and these side-effects contribute substantially to medical costs. Therefore, numerous studies have tried to develop algorithms for predicting treatment response in order to identify those patients who would benefit from treatment. These studies have unravelled a number of host and viral factors that are

Role of sinusoidal endothelial cells of the liver in concanavalin A-induced hepatic injury in mice

Hepatology, Oct 1, 1996

that infiltrating T cells are activated and produce inflamma-CD4 / T lymphocytes have been identi... more that infiltrating T cells are activated and produce inflamma-CD4 / T lymphocytes have been identified as being retory lymphokines. 3,4 In experimental autoimmune hepatitis, sponsible for organ damage in the murine model of ex-T-cell-mediated liver damage could be demonstrated. 5 Howperimental liver injury induced by intravenous injection ever, the early mechanisms leading to liver injury are poorly of concanavalin A (Con A). Liver sinusoidal endothelial characterized. Recruitment of antigen-specific and bystander cells (SEC) and Kupffer's cells (KC) are among the first T cells into the liver and the initiation of the inflammatory cells that come into contact with lymphocytes in the process are poorly understood. liver sinusoid. We aimed to investigate the respective The recently described model of concanavalin A (Con A)role of these cell populations in the initial steps of Tcell-mediated liver injury in Con A-induced hepatitis. induced experimental liver injury in mice has been shown to By electron microscopy, we could show that intravebe mediated by activated T cells. 6 Con A is a T-lymphocyte nously applied Con A bound predominantly to SEC but mitogen in vitro that leads to production of cytokines (innot to KC. KC depletion by gadolinium chloride treatterleukin-2, tumor necrosis factor a [TNF-a], interferon ment of mice did not result in protection from liver ingamma) and lymphocyte proliferation. 7 Eight hours after injury, indicating that KCs are not primarily involved in travenous injection of Con A, a fulminant hepatitis develops the generation of liver injury. We could show that a CD4 / in mice with no other organs being affected. The organ speci-T-cell line (LNC.2) displayed selective cytotoxicity toficity is attributed to the preferential binding of Con A in the ward SEC (ú50%) but not KC (12%) or fibroblasts (5%) in liver. 8 Both the presence and the functional integrity of T the presence of Con A in vitro. Microscopic observation cells are required for Con A-induced hepatitis because imrevealed that the SEC monolayer was rapidly destroyed munodeficient mice (SCID or athymic mice) or mice treated by LnC 2 in the presence of Con A. Specificity of the Con with immunosuppressive drugs such as cyclosporin, FK506, A-induced cytotoxicity was shown by the ability of a or corticosteroids do not develop hepatitis after Con A chalcompetitive ligand, methyl-aD -mannopyranoside, to relenge. 6 By cell-depletion experiments, CD4 / T cells were idenduce T-cell-mediated cytotoxicity to SEC by more than tified as effector cells. 6 TNF-a is a cytokine produced early 50%. Tumor necrosis factor a (TNF-a) was produced by in the course of Con A-induced hepatitis, and blockade of LnC 2 in high amounts after Con A stimulation (ú6 ng/ TNF-a action by antiserum was shown to protect against mL), but antiserum to TNF-a did not reduce LnC 2-medidevelopment of liver injury. 8,9 ated cytotoxicity toward SEC. In conclusion, we could The aim of the present study was to characterize the differshow for the first time that liver SECs have accessory ential role of sinusoidal endothelial cells and Kupffer's cells function and are selectively destroyed by CD4 / T lym-(KCs) in the initial steps of T-cell-mediated liver injury in phocytes in the presence of Con A. We speculate that the model of Con A hepatitis. SEC damage is an early event in T-cell-mediated liver injury recruiting T lymphocytes from the sinusoidal cir-MATERIALS AND METHODS culation. Loss of the SEC barrier function then exposes underlying hepatocytes to further attack by activated T Reagents. All lectins (Con A, Wheat-Germ-Agglutinin I [WGA I], lymphocytes. These results offer a model of initiating Ulex-Europä us I [UEA I]), whether native/fluorescein isothiocyaevents in T-cell-mediated liver diseases, such as viral or nate-labeled or Au-labeled, were purchased from Sigma (Mü nchen, autoimmune hepatitis, and suggest an important role Germany). Gadolinium chloride (GdCl 3) came from Aldrich (Steinheim, Germany). Lipopolysaccharide (B55:50) S. minnesota for sinusoidal endothelial cells. (HEPATOLOGY 1996;24: was from Sigma. The monoclonal antibodies for TNF-a were from 824-829.) Pharmingen (San Diego, CA), and a specific sandwich enzyme-linked Activated T lymphocytes appear to be responsible for organ immunosorbent assay was performed according to the manufacturer's instructions. Antiserum to TNF-a was kindly provided by Prof. damage in chronic viral hepatitis and autoimmune liver dis-Dr. Wendel, Konstanz. Recombinant TNF-a was supplied by Dianova ease. They are found as infiltrating cells in the periportal (Hamburg, Germany). 51 Chromium was purchased from Amersham area in all forms of chronic active hepatitis. 1,2 It was shown (Braunschweig, Germany). Methyl-aD -mannopyranoside was purchased from ICN Flow (Meckenheim, Germany). Sheep red blood cells were obtained from BAG-Hessen (Lich, Ger-Abbreviations: Con A, concanavalin A; TNF-a, tumor necrosis factor a; KC, Kupffer's many), and antiserum to sheep red blood cells was obtained from cell; WGA I, Wheat-Germ-Agglutinin I; UEA I, Ulex Europä us I; FITC, fluorescein isothiocyanate; SEC, sinusoidal endothelial cell; PBS, phosphate-buffered saline. Camon (Wiesbaden, Germany).

Quantification and genotyping of serum HCV-RNA in patients with chronic hepatitis C undergoing interferon treatment

Archives of Virology, Mar 1, 1997

Quantification of serum HCV-RNA and HCV genotyping was studied in 27 patients with chronic hepati... more Quantification of serum HCV-RNA and HCV genotyping was studied in 27 patients with chronic hepatitis C undergoing interferon treatment. Pretreatment serum HCV-RNA levels were quantified using competitive RT-PCR and compared to a quantitative RT-PCR assay based on co-amplification of HCV-RNA with a synthetic RNA standard. HCV genotyping was performed using a line probe reversed hybridisation assay or direct solid-phase sequencing. This study shows the feasibility of performing HCV-RNA quantification. RT-PCR based on co-amplification HCV-RNA titer less than 6;10 genome equivalents/ml serum did correlate with a complete sustained response to interferon in chronic hepatitis C. HCV genotype 1b was predominantly associated with a high non-responder rate. Future prospective trials will be required to evaluate quantitative HCV-RNA levels and HCV genotyping as response predicting parameters for interferon-treatment.

MHC class II genes influence the susceptibility to chronic active hepatitis C

Journal of Hepatology, Aug 1, 1997

Background/Aims: Chronic hepatitis C develops in more than 70% of hepatitis C virus infected subj... more Background/Aims: Chronic hepatitis C develops in more than 70% of hepatitis C virus infected subjects. Viral factors influence the disease course, but little is known about the importance of host factors. Methods: Frequencies of major histocompatibility complex (MHC) class I and class II antigens were analyzed in two groups of patients with chronic hepatitis C virus infection and in control subjects. MHC class I typing was done by standard microlymphocytotoxicity assays. DRBl and DQAl genotyping was done by PCR based typing methods. Results: DRB1*0301 was found in 26 of 75 patients with chronic hepatitis C virus infection (34.7%) and in 12 of 101 control subjects (11.9%) (relative risk 3.9; p<O.OOl). Homozygosity for this allele appeared to confer a stronger risk. In contrast, DRB1*1301 was detected in three subjects with persistent infection HRONIC C infection with the hepatitis C virus (HCV) is a major health problem worldwide. In the United States 3.5 million people are believed to suffer from the disease (1). Chronic liver disease occurs in half of the HCV infected subjects. In a considerable proportion of patients chronic hepatitis may eventually progress to cirrhosis and hepatocellular carcinoma (2-4). Attempts to explain the clinical expression and the behaviour of chronic hepatitis C by viral factors have shown the importance of the viral genotypes and the level of viraemia for the clinical presentation (5,6). However, there remain large inconsistencies, and it is very likely that the immune response of the host can modify disease outcome. Clearance of the virus in

Induction of cytokine production in naive CD4+ T cells by antigen-presenting murine liver sinusoidal endothelial cells but failure to induce differentiation toward Th1 cells☆, ☆☆

Gastroenterology, Jun 1, 1999

We provide evidence that murine LSECs are mature and efficient APCs that induce cytokine expressi... more We provide evidence that murine LSECs are mature and efficient APCs that induce cytokine expression in naive CD4 ϩ T cells. However, LSECs in contrast to conventional APC populations failed to induce differentiation of naive CD4 ϩ T cells toward a T h1 phenotype but promoted development of T h0 cells. The failure of LSECs to induce differentiation toward T h1 together with the production of negative immunomodulatory cytokines (interleukin [IL]-4 and IL-10) by LSEC-primed T cells upon restimulation may contribute to the unique regulation of the immune response in the liver. Materials and Methods Materials Antibodies (R46A2 and AN17.18; American Type Culture Collection) used for interferon (IFN)-␥ sandwich enzyme-linked immunosorbent assay (ELISA) were purified from hybridoma cell culture supernatant following ammonium sulfate precipitation and affinity chromatography on protein G. Similarly, antibodies against IL-4 (clone BVD4-1D11 and clone BVD6-24G2; American Type Culture Collection) were purified from hybridoma culture supernatants. Recombinant IL-12 and antibodies against IL-12 p35/40 heterodimer were provided by Dr. M. Gately (Hoffmann La-Roche, Nutley, CT). Recombinant IL-18 was purchased from Peprotech (London, England). Goat antiserum to rat immunoglobulin (Ig) as well as rat Ig was obtained from Dianova (Hamburg, Germany). Recombinant mouse IFN-␥, IL-10, IL-4, and monoclonal antibody to CD3⑀ (clone 17A2) were obtained from Pharmingen (Hamburg, Germany). Monoclonal antibodies for detection of IL-10 were obtained from Pharmingen (clone JES5-2A5 and clone JES5-16E3). Heat-killed Listeria monocytogenes were a gift from J. Szeliga (Institute Microbiology and Immunology, University of Lodz, Poland). Pansorbin (SAC) was purchased from Calbiochem (La Jolla, CA). Monoclonal antibody to I-E d (P7/7) was kindly provided by B. Arnold, Deutsches Krebsforschungszentrum Heidelberg (Germany). Cell Isolation and Culture Murine sinusoidal endothelial cells and Kupffer cells.

Local control of the immune response in the liver

Immunological Reviews, Apr 1, 2000

Development and functional consequences of LPS tolerance in sinusoidal endothelial cells of the liver

Journal of Leukocyte Biology, May 1, 2005

Kupffer cells and liver sinusoidal endothelial cells (LSEC) clear portal venous blood from gut-de... more Kupffer cells and liver sinusoidal endothelial cells (LSEC) clear portal venous blood from gut-derived bacterial degradation products such as lipopolysaccharide (LPS) without inducing a local inflammatory reaction. LPS tolerance was reported for Kupffer cells, but little is known whether sensitivity of LSEC toward LPS is dynamically regulated. Here, we demonstrate that LSEC react to LPS directly as a function of constitutive Toll-like receptor 4 (TLR4)/CD14 expression but gain a LPS-refractory state upon repetitive stimulation without loss of scavenger activity. LPS tolerance in LSEC is characterized by reduced nuclear localization of nuclear factor-B upon LPS rechallenge. In contrast to monocytes, however, TLR4 surface expression of LSEC is not altered by LPS stimulation and thus does not account for LPS tolerance. Mechanistically, LPS tolerance in LSEC is linked to prostanoid production and may account for cross-tolerance of LPS-treated LSEC to interferon-␥ stimulation. Functionally, LPS tolerance in LSEC results in reduced leukocyte adhesion following LPS rechallenge as a consequence of decreased CD54 surface expression. Furthermore, LPS tolerance is operative in vivo, as we observed by intravital microscopy-reduced leukocyte adhesion to LSEC and improved sinusoidal microcirculation in the liver after repetitive LPS challenges. Our results support the notion that LPS tolerance in organ-resident scavenger LSEC contributes to local hepatic control of inflammation.

Complete congenital heart block in autoimmune hepatitis (SLA-positive)

Journal of Hepatology, 1994

Complete congenital heart block is a serious complication of neonatal lupus erythematosus which m... more Complete congenital heart block is a serious complication of neonatal lupus erythematosus which most often occurs in children of mothers suffering from connective tissue disease. We report the occurrence of complete congenital heart block associated with autoimmune hepatitis (SLA-positive). A 32-year-old woman was treated for more than 10 years for autoimmune hepatitis (SLA-/ANA-positive) and remained in clinical remission under immunosuppressive therapy. She showed an MHC-haplotype typical for autoimmune hepatitis (A1, B8, DR3). After a normal first pregnancy, an emergency caesarean section was performed in the 32nd week of her second pregnancy because of fetal bradycardia. The child died a few hours after delivery of complete congenital AV-block. Retrospective analysis of the maternal serum showed the emergence of SS-A/Ro-antibodies prior to the second pregnancy. The maternal serum antibodies were reactive with the 52 kD SS-A/Ro-antigen, as demonstrated by immunoblot employing recombinant SSA/Ro-antigen. The occurrence of complete congenital heart block has been shown to be associated with the presence of SS-A/Ro antibodies as well as the MHC-haplotype DR3. With respect to this genetic linkage, pregnant patients with autoimmune hepatitis and the MHC-haplotype DR3 should be examined for the presence of SS-A/Ro-antibodies. They should be closely followed during pregnancy to enable early detection of the development of congenital heart block, as prevention by plasmapheresis plus dexamethasone may be possible at an early stage.

Endotoxin Down-Regulates T Cell Activation by Antigen-Presenting Liver Sinusoidal Endothelial Cells

The Journal of Immunology

Endotoxin is physiologically present in portal venous blood at concentrations of 100 pg/ml to 1 n... more Endotoxin is physiologically present in portal venous blood at concentrations of 100 pg/ml to 1 ng/ml. Clearance of endotoxin from portal blood occurs through sinusoidal lining cells, i.e., Kupffer cells, and liver sinusoidal endothelial cells (LSEC). We have recently shown that LSEC are fully efficient APCs. Here, we studied the influence of endotoxin on the accessory function of LSEC. Incubation of Ag-presenting LSEC with physiological concentrations of endotoxin lead to ≥80% reduction of the accessory function, measured by release of IFN-γ from CD4+ T cells. In contrast, conventional APC populations rather showed an increase of the accessory function after endotoxin treatment. Inhibition of the accessory function in LSEC by endotoxin was not due to lack of soluble costimulatory signals, because neither supplemental IL-1β, IL-2, IFN-γ, or IL-12 could rescue the accessory function. Ag uptake was not influenced by endotoxin in LSEC. However, we found that endotoxin led to alkaliniza...

Viral and host factors in the prediction of response to interferon‐α therapy in chronic hepatitis C after long‐term follow‐up

Journal of Viral Hepatitis, 1998

Acute infection with hepatitis C virus (HCV) develops into a chronic hepatitis in about 50–70% of... more Acute infection with hepatitis C virus (HCV) develops into a chronic hepatitis in about 50–70% of patients. Treatment of these patients with interferon‐α (IFN‐α) results in a sustained long‐term response in only 15–20% but causes numerous unwanted side‐effects in a higher percentage of patients. The aim of our study was to define host or viral parameters that would allow identification of responders and non‐responders to IFN‐α prior to the onset of treatment. We studied a group of 87 patients suffering from chronic hepatitis C who were treated with IFN‐α. After long‐term follow‐up, 18 patients (21%) showed a sustained response to IFN‐α therapy (normalization of serum transaminases and loss of viral RNA from serum) for up to 7 years after therapy had ceased. By univariate and multivariate analyses, no host factors were found to be predictive of response to therapy. Neither the degree of inflammation or fibrosis in liver biopsy samples obtained before treatment nor immunogenetic facto...

Ligase chain reaction (LCR®) assay for semi‐quantitative detection of HBV DNA in mononuclear leukocytes of patients with chronic hepatitis B

Journal of Viral Hepatitis, 1996

Summary. A ligase chain reaction (LCR®)‐based approach to detect hepatitis B virus (HBV) DNA in p... more Summary. A ligase chain reaction (LCR®)‐based approach to detect hepatitis B virus (HBV) DNA in peripheral blood mononuclear cells (PBMC) is described. Using this new amplification technique, we determined semi‐quantitatively the amount of a short HBV S‐gene fragment in PBMC lysates of 25 patients with different forms of chronic hepatitis (group A (n= 8), hepatitis B s antigen (HBsAg)+/hepatitis B e antigen (HBeAg)+; group B (n= 9), HBsAg+/HBeAg‐; group C (n= 8), HBsAg‐/HBeAg‐). The LCR results were compared with the findings obtained with polymerase chain reaction (PCR) amplification of three distinct HBV gene regions (preS1/2, S and C) and related to the serological profiles of the patients. Depending on the primer pair used for PCR amplification, sensitivity of HBV LCR in PBMC was equivalent or slightly superior to PCR. The highest positivity rate for HBV DNA was observed in the HBeAg+ and HBV DNA seropositive group (8/8) and was lower in the other patient groups B (4/9) and C (1...

Sustained elimination of hepatitis B virus from serum induced in a patient with chronic hepatitis B and advanced human immunodeficiency virus infection

The Clinical Investigator, 1994

A 48-year-old male patient was admitted with acquired immunodeficiency syndrome (stage III, Cente... more A 48-year-old male patient was admitted with acquired immunodeficiency syndrome (stage III, Centers for Disease Control 1993) and viremic hepatitis B. Blood CD4 count was 15/microliters. Discontinuation of prednisolone, previously prescribed by the patient&amp;amp;#39;s family practitioner because of elevated liver enzymes, resulted in severe hepatitis (alanine aminotransferase &amp;amp;gt; 300U/l). Administration of interferon-alpha (9 x 10(6) U s.c. 3 x weekly) was initiated. Serum markers of viral replication disappeared, and aminotransferase levels returned to normal within a few weeks. The patient&amp;amp;#39;s serum was found negative for HBsAg after 3 months. Immunohistochemical analysis of liver biopsies before and during interferon therapy showed disappearance of all hepatitis B virus antigens and a marked reduction in inflammatory activity. Hepatitis B virus seroconversion remained stable until the patient died from the syndrome 2 years later. This case shows that in spite of severe HIV-associated immune deficiency with CD4 counts constantly below 100/microliters, interferon-alpha can lead to sustained serological and histological improvement of viremic hepatitis B. Previous administration and discontinuation of cortisone may have helped to reach this effect.

Development and functional consequences of LPS tolerance in sinusoidal endothelial cells of the liver

Journal of Leukocyte Biology, 2005

Kupffer cells and liver sinusoidal endothelial cells (LSEC) clear portal venous blood from gut-de... more Kupffer cells and liver sinusoidal endothelial cells (LSEC) clear portal venous blood from gut-derived bacterial degradation products such as lipopolysaccharide (LPS) without inducing a local inflammatory reaction. LPS tolerance was reported for Kupffer cells, but little is known whether sensitivity of LSEC toward LPS is dynamically regulated. Here, we demonstrate that LSEC react to LPS directly as a function of constitutive Toll-like receptor 4 (TLR4)/CD14 expression but gain a LPS-refractory state upon repetitive stimulation without loss of scavenger activity. LPS tolerance in LSEC is characterized by reduced nuclear localization of nuclear factor-κB upon LPS rechallenge. In contrast to monocytes, however, TLR4 surface expression of LSEC is not altered by LPS stimulation and thus does not account for LPS tolerance. Mechanistically, LPS tolerance in LSEC is linked to prostanoid production and may account for cross-tolerance of LPS-treated LSEC to interferon-γ stimulation. Functiona...

MHC class II genes influence the susceptibility to chronic active hepatitis C

Journal of Hepatology, 1997

Background/Aims: Chronic hepatitis C develops in more than 70% of hepatitis C virus infected subj... more Background/Aims: Chronic hepatitis C develops in more than 70% of hepatitis C virus infected subjects. Viral factors influence the disease course, but little is known about the importance of host factors. Methods: Frequencies of major histocompatibility complex (MHC) class I and class II antigens were analyzed in two groups of patients with chronic hepatitis C virus infection and in control subjects. MHC class I typing was done by standard microlymphocytotoxicity assays. DRBl and DQAl genotyping was done by PCR based typing methods. Results: DRB1*0301 was found in 26 of 75 patients with chronic hepatitis C virus infection (34.7%) and in 12 of 101 control subjects (11.9%) (relative risk 3.9; p<O.OOl). Homozygosity for this allele appeared to confer a stronger risk. In contrast, DRB1*1301 was detected in three subjects with persistent infection HRONIC C infection with the hepatitis C virus (HCV) is a major health problem worldwide. In the United States 3.5 million people are believed to suffer from the disease (1). Chronic liver disease occurs in half of the HCV infected subjects. In a considerable proportion of patients chronic hepatitis may eventually progress to cirrhosis and hepatocellular carcinoma (2-4). Attempts to explain the clinical expression and the behaviour of chronic hepatitis C by viral factors have shown the importance of the viral genotypes and the level of viraemia for the clinical presentation (5,6). However, there remain large inconsistencies, and it is very likely that the immune response of the host can modify disease outcome. Clearance of the virus in

Human Kupffer cells secrete IL-10 in response to lipopolysaccharide (LPS) challenge

Journal of Hepatology, 1995

Kupffer cells are involved in local immunoregulation in the liver by secretion of cytokines and d... more Kupffer cells are involved in local immunoregulation in the liver by secretion of cytokines and direct cellular contact. They are able to influence the function of other liver cells, i.e. sinusoidal endothelial cells, Ito cells and hepatocytes. The three known major functions of Kupffer cells are clearance of endotoxin from the portal circulation, release of soluble mediators and presentation of antigen. Human Kupffer cells were isolated by collagenase perfusion followed by centrifugal elutriation and analyzed for cytokine secretion after 3 days in culture. We found that freshly isolated human Kupffer cells secreted the anti-inflammatory cytokine interleukin-10 in response to stimulation with lipopolysaccharide. The release of interleukin-10 was maximal 12-24 h after lipopolysaccharide challenge. Furthermore, we could show that exogenous interleukin-10 downregulated the release of the proinflammatory cytokines interleukin-6 and tumor necrosis factor alpha by Kupffer cells after lipopolysaccharide stimulation. The release of interleukin-6 was maximal 24 h after stimulation and interleukin-10 inhibited interleukin-6 release after 6 h. Tumor necrosis factor alpha showed maximal secretion 6 h after stimulation and exogenous interleukin-10 also downregulated the tumor necrosis factor alpha release after 6 h. Kupffer cells are exposed physiologically to lipopolysaccharide present in portal venous blood. Given the known anti-inflammatory effect of interleukin-10, our findings of secretion of interleukin-10 by Kupffer cells in response to lipopolysaccharide and suppression of interleukin-6 and tumor necrosis factor alpha release by Kupffer cells in vitro through exogenous interleukin-10 suggest an important role for interleukin-10 in the regulation of the local immune response in the liver sinusoid.

Regulation of chemokine (MCP-1, IP-10, MIP-2) secretion in liver sinusoidal endothelial cell by endotoxin, IL-10 and IL-4

Immunology Letters, 1997

Liver sinusoidal endothelial cells prime naive T cells and induce a Th2-shift implication for induction of tolerance in the liver

Immunology Letters, 1997

Interleukin-10 expression is autoregulated at the transcriptional level in human and murine kupffer cells

Hepatology, 1998

Interleukin 10 (IL-10) is known to downregulate immune responses. The regulation of IL-10 gene ex... more Interleukin 10 (IL-10) is known to downregulate immune responses. The regulation of IL-10 gene expression therefore determines the outcome of local immune reactions. We investigated time course and downregulation of IL-10 production in primary Kupffer's cells (KC), which are known to secrete IL-10 in response to endotoxin challenge. Human and murine KC were isolated by centrifugal elutriation and investigated for IL-10 gene expression by a two-step amplification procedure (reverse transcriptasepolymerase chain reaction [PCR] followed by T7-polymerase chain reaction). We show that IL-10 messenger ribonucleic acid (mRNA) showed a G450 fold increase in KC 2 hours after endotoxin challenge. IL-10 protein release from KC strictly depended on de novo protein synthesis. Endotoxin mediated increase in IL-10 gene expression was downregulated by exogenous (G350-fold reduction of IL-10 mRNA level), as well as endogenous IL-10 protein, showing a negative autoregulatory feedback loop. IL-10 receptor expression was found to be constitutive and functional in KC. Early expression of IL-10 in KC may be of functional relevance to the outcome of immune and inflammatory reactions in the liver sinusoid. The negative autoregulation of IL-10 expression may represent a mechanism to regain a state of functional responsiveness in the microenvironment towards new proinflammatory stimuli. In conclusion, autoregulatory downregulation of IL-10 expression in KC may account for important regulatory steps of local immune response in the liver sinusoid. (HEPATOLOGY 1998;27:93-99.

IL-10 down-regulates T cell activation by antigen-presenting liver sinusoidal endothelial cells through decreased antigen uptake via the mannose receptor and lowered surface expression of accessory molecules

Clinical and Experimental Immunology, Dec 1, 1998

Our study demonstrates that antigen-presenting liver sinusoidal endothelial cells (LSEC) induce p... more Our study demonstrates that antigen-presenting liver sinusoidal endothelial cells (LSEC) induce production of interferon-gamma (IFN-g) from cloned Th1 CD4 þ T cells. We show that LSEC used the mannose receptor for antigen uptake, which further strengthened the role of LSEC as antigenpresenting cell (APC) population in the liver. The ability of LSEC to activate cloned CD4 þ T cells antigen-specifically was down-regulated by exogenous prostaglandin E 2 (PGE 2) and by IL-10. We identify two separate mechanisms by which IL-10 down-regulated T cell activation through LSEC. IL-10 decreased the constitutive surface expression of MHC class II as well as of the accessory molecules CD80 and CD86 on LSEC. Furthermore, IL-10 diminished mannose receptor activity in LSEC. Decreased antigen uptake via the mannose receptor and decreased expression of accessory molecules may explain the down-regulation of T cell activation through IL-10. Importantly, the expression of low numbers of antigen on MHC II in the absence of accessory signals on LSEC may lead to induction of anergy in T cells. Because PGE 2 and IL-10 are released from LSEC or Kupffer cells (KC) in response to those concentrations of endotoxin found physiologically in portal venous blood, it is possible that the continuous presence of these mediators and their negative effect on the local APC may explain the inability of the liver to induce T cell activation and to clear chronic infections. Our results support the notion that antigen presentation by LSEC in the hepatic microenvironment contributes to the observed inability to mount an effective cell-mediated immune response in the liver.

HIV und Schwangerschaft: Risiken der Übertragung und therapeutische Möglichkeiten

Gynakologisch-geburtshilfliche Rundschau, 1996

ABSTRACT

Viral and host factors in the prediction of response to interferon-alpha therapy in chronic hepatitis C after long-term follow-up

Journal of Viral Hepatitis, Nov 1, 1998

Chronic hepatitis caused by hepatitis C virus (HCV) infection is a common health problem affectin... more Chronic hepatitis caused by hepatitis C virus (HCV) infection is a common health problem affecting probably 3% of the world's population. Established treatment of chronic hepatitis C consists mainly of the use of interferon-α (IFN-α), which leads to a sustained treatment response in only 15-20% of patients. IFN-α therapy is frequently accompanied by many undesirable effects, e.g. headache, myalgia, thrombocytopenia or psychosis, and these side-effects contribute substantially to medical costs. Therefore, numerous studies have tried to develop algorithms for predicting treatment response in order to identify those patients who would benefit from treatment. These studies have unravelled a number of host and viral factors that are

Role of sinusoidal endothelial cells of the liver in concanavalin A-induced hepatic injury in mice

Hepatology, Oct 1, 1996

that infiltrating T cells are activated and produce inflamma-CD4 / T lymphocytes have been identi... more that infiltrating T cells are activated and produce inflamma-CD4 / T lymphocytes have been identified as being retory lymphokines. 3,4 In experimental autoimmune hepatitis, sponsible for organ damage in the murine model of ex-T-cell-mediated liver damage could be demonstrated. 5 Howperimental liver injury induced by intravenous injection ever, the early mechanisms leading to liver injury are poorly of concanavalin A (Con A). Liver sinusoidal endothelial characterized. Recruitment of antigen-specific and bystander cells (SEC) and Kupffer's cells (KC) are among the first T cells into the liver and the initiation of the inflammatory cells that come into contact with lymphocytes in the process are poorly understood. liver sinusoid. We aimed to investigate the respective The recently described model of concanavalin A (Con A)role of these cell populations in the initial steps of Tcell-mediated liver injury in Con A-induced hepatitis. induced experimental liver injury in mice has been shown to By electron microscopy, we could show that intravebe mediated by activated T cells. 6 Con A is a T-lymphocyte nously applied Con A bound predominantly to SEC but mitogen in vitro that leads to production of cytokines (innot to KC. KC depletion by gadolinium chloride treatterleukin-2, tumor necrosis factor a [TNF-a], interferon ment of mice did not result in protection from liver ingamma) and lymphocyte proliferation. 7 Eight hours after injury, indicating that KCs are not primarily involved in travenous injection of Con A, a fulminant hepatitis develops the generation of liver injury. We could show that a CD4 / in mice with no other organs being affected. The organ speci-T-cell line (LNC.2) displayed selective cytotoxicity toficity is attributed to the preferential binding of Con A in the ward SEC (ú50%) but not KC (12%) or fibroblasts (5%) in liver. 8 Both the presence and the functional integrity of T the presence of Con A in vitro. Microscopic observation cells are required for Con A-induced hepatitis because imrevealed that the SEC monolayer was rapidly destroyed munodeficient mice (SCID or athymic mice) or mice treated by LnC 2 in the presence of Con A. Specificity of the Con with immunosuppressive drugs such as cyclosporin, FK506, A-induced cytotoxicity was shown by the ability of a or corticosteroids do not develop hepatitis after Con A chalcompetitive ligand, methyl-aD -mannopyranoside, to relenge. 6 By cell-depletion experiments, CD4 / T cells were idenduce T-cell-mediated cytotoxicity to SEC by more than tified as effector cells. 6 TNF-a is a cytokine produced early 50%. Tumor necrosis factor a (TNF-a) was produced by in the course of Con A-induced hepatitis, and blockade of LnC 2 in high amounts after Con A stimulation (ú6 ng/ TNF-a action by antiserum was shown to protect against mL), but antiserum to TNF-a did not reduce LnC 2-medidevelopment of liver injury. 8,9 ated cytotoxicity toward SEC. In conclusion, we could The aim of the present study was to characterize the differshow for the first time that liver SECs have accessory ential role of sinusoidal endothelial cells and Kupffer's cells function and are selectively destroyed by CD4 / T lym-(KCs) in the initial steps of T-cell-mediated liver injury in phocytes in the presence of Con A. We speculate that the model of Con A hepatitis. SEC damage is an early event in T-cell-mediated liver injury recruiting T lymphocytes from the sinusoidal cir-MATERIALS AND METHODS culation. Loss of the SEC barrier function then exposes underlying hepatocytes to further attack by activated T Reagents. All lectins (Con A, Wheat-Germ-Agglutinin I [WGA I], lymphocytes. These results offer a model of initiating Ulex-Europä us I [UEA I]), whether native/fluorescein isothiocyaevents in T-cell-mediated liver diseases, such as viral or nate-labeled or Au-labeled, were purchased from Sigma (Mü nchen, autoimmune hepatitis, and suggest an important role Germany). Gadolinium chloride (GdCl 3) came from Aldrich (Steinheim, Germany). Lipopolysaccharide (B55:50) S. minnesota for sinusoidal endothelial cells. (HEPATOLOGY 1996;24: was from Sigma. The monoclonal antibodies for TNF-a were from 824-829.) Pharmingen (San Diego, CA), and a specific sandwich enzyme-linked Activated T lymphocytes appear to be responsible for organ immunosorbent assay was performed according to the manufacturer's instructions. Antiserum to TNF-a was kindly provided by Prof. damage in chronic viral hepatitis and autoimmune liver dis-Dr. Wendel, Konstanz. Recombinant TNF-a was supplied by Dianova ease. They are found as infiltrating cells in the periportal (Hamburg, Germany). 51 Chromium was purchased from Amersham area in all forms of chronic active hepatitis. 1,2 It was shown (Braunschweig, Germany). Methyl-aD -mannopyranoside was purchased from ICN Flow (Meckenheim, Germany). Sheep red blood cells were obtained from BAG-Hessen (Lich, Ger-Abbreviations: Con A, concanavalin A; TNF-a, tumor necrosis factor a; KC, Kupffer's many), and antiserum to sheep red blood cells was obtained from cell; WGA I, Wheat-Germ-Agglutinin I; UEA I, Ulex Europä us I; FITC, fluorescein isothiocyanate; SEC, sinusoidal endothelial cell; PBS, phosphate-buffered saline. Camon (Wiesbaden, Germany).

Quantification and genotyping of serum HCV-RNA in patients with chronic hepatitis C undergoing interferon treatment

Archives of Virology, Mar 1, 1997

Quantification of serum HCV-RNA and HCV genotyping was studied in 27 patients with chronic hepati... more Quantification of serum HCV-RNA and HCV genotyping was studied in 27 patients with chronic hepatitis C undergoing interferon treatment. Pretreatment serum HCV-RNA levels were quantified using competitive RT-PCR and compared to a quantitative RT-PCR assay based on co-amplification of HCV-RNA with a synthetic RNA standard. HCV genotyping was performed using a line probe reversed hybridisation assay or direct solid-phase sequencing. This study shows the feasibility of performing HCV-RNA quantification. RT-PCR based on co-amplification HCV-RNA titer less than 6;10 genome equivalents/ml serum did correlate with a complete sustained response to interferon in chronic hepatitis C. HCV genotype 1b was predominantly associated with a high non-responder rate. Future prospective trials will be required to evaluate quantitative HCV-RNA levels and HCV genotyping as response predicting parameters for interferon-treatment.

MHC class II genes influence the susceptibility to chronic active hepatitis C

Journal of Hepatology, Aug 1, 1997

Background/Aims: Chronic hepatitis C develops in more than 70% of hepatitis C virus infected subj... more Background/Aims: Chronic hepatitis C develops in more than 70% of hepatitis C virus infected subjects. Viral factors influence the disease course, but little is known about the importance of host factors. Methods: Frequencies of major histocompatibility complex (MHC) class I and class II antigens were analyzed in two groups of patients with chronic hepatitis C virus infection and in control subjects. MHC class I typing was done by standard microlymphocytotoxicity assays. DRBl and DQAl genotyping was done by PCR based typing methods. Results: DRB1*0301 was found in 26 of 75 patients with chronic hepatitis C virus infection (34.7%) and in 12 of 101 control subjects (11.9%) (relative risk 3.9; p<O.OOl). Homozygosity for this allele appeared to confer a stronger risk. In contrast, DRB1*1301 was detected in three subjects with persistent infection HRONIC C infection with the hepatitis C virus (HCV) is a major health problem worldwide. In the United States 3.5 million people are believed to suffer from the disease (1). Chronic liver disease occurs in half of the HCV infected subjects. In a considerable proportion of patients chronic hepatitis may eventually progress to cirrhosis and hepatocellular carcinoma (2-4). Attempts to explain the clinical expression and the behaviour of chronic hepatitis C by viral factors have shown the importance of the viral genotypes and the level of viraemia for the clinical presentation (5,6). However, there remain large inconsistencies, and it is very likely that the immune response of the host can modify disease outcome. Clearance of the virus in

Induction of cytokine production in naive CD4+ T cells by antigen-presenting murine liver sinusoidal endothelial cells but failure to induce differentiation toward Th1 cells☆, ☆☆

Gastroenterology, Jun 1, 1999

We provide evidence that murine LSECs are mature and efficient APCs that induce cytokine expressi... more We provide evidence that murine LSECs are mature and efficient APCs that induce cytokine expression in naive CD4 ϩ T cells. However, LSECs in contrast to conventional APC populations failed to induce differentiation of naive CD4 ϩ T cells toward a T h1 phenotype but promoted development of T h0 cells. The failure of LSECs to induce differentiation toward T h1 together with the production of negative immunomodulatory cytokines (interleukin [IL]-4 and IL-10) by LSEC-primed T cells upon restimulation may contribute to the unique regulation of the immune response in the liver. Materials and Methods Materials Antibodies (R46A2 and AN17.18; American Type Culture Collection) used for interferon (IFN)-␥ sandwich enzyme-linked immunosorbent assay (ELISA) were purified from hybridoma cell culture supernatant following ammonium sulfate precipitation and affinity chromatography on protein G. Similarly, antibodies against IL-4 (clone BVD4-1D11 and clone BVD6-24G2; American Type Culture Collection) were purified from hybridoma culture supernatants. Recombinant IL-12 and antibodies against IL-12 p35/40 heterodimer were provided by Dr. M. Gately (Hoffmann La-Roche, Nutley, CT). Recombinant IL-18 was purchased from Peprotech (London, England). Goat antiserum to rat immunoglobulin (Ig) as well as rat Ig was obtained from Dianova (Hamburg, Germany). Recombinant mouse IFN-␥, IL-10, IL-4, and monoclonal antibody to CD3⑀ (clone 17A2) were obtained from Pharmingen (Hamburg, Germany). Monoclonal antibodies for detection of IL-10 were obtained from Pharmingen (clone JES5-2A5 and clone JES5-16E3). Heat-killed Listeria monocytogenes were a gift from J. Szeliga (Institute Microbiology and Immunology, University of Lodz, Poland). Pansorbin (SAC) was purchased from Calbiochem (La Jolla, CA). Monoclonal antibody to I-E d (P7/7) was kindly provided by B. Arnold, Deutsches Krebsforschungszentrum Heidelberg (Germany). Cell Isolation and Culture Murine sinusoidal endothelial cells and Kupffer cells.

Local control of the immune response in the liver

Immunological Reviews, Apr 1, 2000

Development and functional consequences of LPS tolerance in sinusoidal endothelial cells of the liver

Journal of Leukocyte Biology, May 1, 2005

Kupffer cells and liver sinusoidal endothelial cells (LSEC) clear portal venous blood from gut-de... more Kupffer cells and liver sinusoidal endothelial cells (LSEC) clear portal venous blood from gut-derived bacterial degradation products such as lipopolysaccharide (LPS) without inducing a local inflammatory reaction. LPS tolerance was reported for Kupffer cells, but little is known whether sensitivity of LSEC toward LPS is dynamically regulated. Here, we demonstrate that LSEC react to LPS directly as a function of constitutive Toll-like receptor 4 (TLR4)/CD14 expression but gain a LPS-refractory state upon repetitive stimulation without loss of scavenger activity. LPS tolerance in LSEC is characterized by reduced nuclear localization of nuclear factor-B upon LPS rechallenge. In contrast to monocytes, however, TLR4 surface expression of LSEC is not altered by LPS stimulation and thus does not account for LPS tolerance. Mechanistically, LPS tolerance in LSEC is linked to prostanoid production and may account for cross-tolerance of LPS-treated LSEC to interferon-␥ stimulation. Functionally, LPS tolerance in LSEC results in reduced leukocyte adhesion following LPS rechallenge as a consequence of decreased CD54 surface expression. Furthermore, LPS tolerance is operative in vivo, as we observed by intravital microscopy-reduced leukocyte adhesion to LSEC and improved sinusoidal microcirculation in the liver after repetitive LPS challenges. Our results support the notion that LPS tolerance in organ-resident scavenger LSEC contributes to local hepatic control of inflammation.

Complete congenital heart block in autoimmune hepatitis (SLA-positive)

Journal of Hepatology, 1994

Complete congenital heart block is a serious complication of neonatal lupus erythematosus which m... more Complete congenital heart block is a serious complication of neonatal lupus erythematosus which most often occurs in children of mothers suffering from connective tissue disease. We report the occurrence of complete congenital heart block associated with autoimmune hepatitis (SLA-positive). A 32-year-old woman was treated for more than 10 years for autoimmune hepatitis (SLA-/ANA-positive) and remained in clinical remission under immunosuppressive therapy. She showed an MHC-haplotype typical for autoimmune hepatitis (A1, B8, DR3). After a normal first pregnancy, an emergency caesarean section was performed in the 32nd week of her second pregnancy because of fetal bradycardia. The child died a few hours after delivery of complete congenital AV-block. Retrospective analysis of the maternal serum showed the emergence of SS-A/Ro-antibodies prior to the second pregnancy. The maternal serum antibodies were reactive with the 52 kD SS-A/Ro-antigen, as demonstrated by immunoblot employing recombinant SSA/Ro-antigen. The occurrence of complete congenital heart block has been shown to be associated with the presence of SS-A/Ro antibodies as well as the MHC-haplotype DR3. With respect to this genetic linkage, pregnant patients with autoimmune hepatitis and the MHC-haplotype DR3 should be examined for the presence of SS-A/Ro-antibodies. They should be closely followed during pregnancy to enable early detection of the development of congenital heart block, as prevention by plasmapheresis plus dexamethasone may be possible at an early stage.

Endotoxin Down-Regulates T Cell Activation by Antigen-Presenting Liver Sinusoidal Endothelial Cells

The Journal of Immunology

Endotoxin is physiologically present in portal venous blood at concentrations of 100 pg/ml to 1 n... more Endotoxin is physiologically present in portal venous blood at concentrations of 100 pg/ml to 1 ng/ml. Clearance of endotoxin from portal blood occurs through sinusoidal lining cells, i.e., Kupffer cells, and liver sinusoidal endothelial cells (LSEC). We have recently shown that LSEC are fully efficient APCs. Here, we studied the influence of endotoxin on the accessory function of LSEC. Incubation of Ag-presenting LSEC with physiological concentrations of endotoxin lead to ≥80% reduction of the accessory function, measured by release of IFN-γ from CD4+ T cells. In contrast, conventional APC populations rather showed an increase of the accessory function after endotoxin treatment. Inhibition of the accessory function in LSEC by endotoxin was not due to lack of soluble costimulatory signals, because neither supplemental IL-1β, IL-2, IFN-γ, or IL-12 could rescue the accessory function. Ag uptake was not influenced by endotoxin in LSEC. However, we found that endotoxin led to alkaliniza...

Viral and host factors in the prediction of response to interferon‐α therapy in chronic hepatitis C after long‐term follow‐up

Journal of Viral Hepatitis, 1998

Acute infection with hepatitis C virus (HCV) develops into a chronic hepatitis in about 50–70% of... more Acute infection with hepatitis C virus (HCV) develops into a chronic hepatitis in about 50–70% of patients. Treatment of these patients with interferon‐α (IFN‐α) results in a sustained long‐term response in only 15–20% but causes numerous unwanted side‐effects in a higher percentage of patients. The aim of our study was to define host or viral parameters that would allow identification of responders and non‐responders to IFN‐α prior to the onset of treatment. We studied a group of 87 patients suffering from chronic hepatitis C who were treated with IFN‐α. After long‐term follow‐up, 18 patients (21%) showed a sustained response to IFN‐α therapy (normalization of serum transaminases and loss of viral RNA from serum) for up to 7 years after therapy had ceased. By univariate and multivariate analyses, no host factors were found to be predictive of response to therapy. Neither the degree of inflammation or fibrosis in liver biopsy samples obtained before treatment nor immunogenetic facto...

Ligase chain reaction (LCR®) assay for semi‐quantitative detection of HBV DNA in mononuclear leukocytes of patients with chronic hepatitis B

Journal of Viral Hepatitis, 1996

Summary. A ligase chain reaction (LCR®)‐based approach to detect hepatitis B virus (HBV) DNA in p... more Summary. A ligase chain reaction (LCR®)‐based approach to detect hepatitis B virus (HBV) DNA in peripheral blood mononuclear cells (PBMC) is described. Using this new amplification technique, we determined semi‐quantitatively the amount of a short HBV S‐gene fragment in PBMC lysates of 25 patients with different forms of chronic hepatitis (group A (n= 8), hepatitis B s antigen (HBsAg)+/hepatitis B e antigen (HBeAg)+; group B (n= 9), HBsAg+/HBeAg‐; group C (n= 8), HBsAg‐/HBeAg‐). The LCR results were compared with the findings obtained with polymerase chain reaction (PCR) amplification of three distinct HBV gene regions (preS1/2, S and C) and related to the serological profiles of the patients. Depending on the primer pair used for PCR amplification, sensitivity of HBV LCR in PBMC was equivalent or slightly superior to PCR. The highest positivity rate for HBV DNA was observed in the HBeAg+ and HBV DNA seropositive group (8/8) and was lower in the other patient groups B (4/9) and C (1...

Sustained elimination of hepatitis B virus from serum induced in a patient with chronic hepatitis B and advanced human immunodeficiency virus infection

The Clinical Investigator, 1994

A 48-year-old male patient was admitted with acquired immunodeficiency syndrome (stage III, Cente... more A 48-year-old male patient was admitted with acquired immunodeficiency syndrome (stage III, Centers for Disease Control 1993) and viremic hepatitis B. Blood CD4 count was 15/microliters. Discontinuation of prednisolone, previously prescribed by the patient&amp;amp;#39;s family practitioner because of elevated liver enzymes, resulted in severe hepatitis (alanine aminotransferase &amp;amp;gt; 300U/l). Administration of interferon-alpha (9 x 10(6) U s.c. 3 x weekly) was initiated. Serum markers of viral replication disappeared, and aminotransferase levels returned to normal within a few weeks. The patient&amp;amp;#39;s serum was found negative for HBsAg after 3 months. Immunohistochemical analysis of liver biopsies before and during interferon therapy showed disappearance of all hepatitis B virus antigens and a marked reduction in inflammatory activity. Hepatitis B virus seroconversion remained stable until the patient died from the syndrome 2 years later. This case shows that in spite of severe HIV-associated immune deficiency with CD4 counts constantly below 100/microliters, interferon-alpha can lead to sustained serological and histological improvement of viremic hepatitis B. Previous administration and discontinuation of cortisone may have helped to reach this effect.

Development and functional consequences of LPS tolerance in sinusoidal endothelial cells of the liver

Journal of Leukocyte Biology, 2005

Kupffer cells and liver sinusoidal endothelial cells (LSEC) clear portal venous blood from gut-de... more Kupffer cells and liver sinusoidal endothelial cells (LSEC) clear portal venous blood from gut-derived bacterial degradation products such as lipopolysaccharide (LPS) without inducing a local inflammatory reaction. LPS tolerance was reported for Kupffer cells, but little is known whether sensitivity of LSEC toward LPS is dynamically regulated. Here, we demonstrate that LSEC react to LPS directly as a function of constitutive Toll-like receptor 4 (TLR4)/CD14 expression but gain a LPS-refractory state upon repetitive stimulation without loss of scavenger activity. LPS tolerance in LSEC is characterized by reduced nuclear localization of nuclear factor-κB upon LPS rechallenge. In contrast to monocytes, however, TLR4 surface expression of LSEC is not altered by LPS stimulation and thus does not account for LPS tolerance. Mechanistically, LPS tolerance in LSEC is linked to prostanoid production and may account for cross-tolerance of LPS-treated LSEC to interferon-γ stimulation. Functiona...

MHC class II genes influence the susceptibility to chronic active hepatitis C

Journal of Hepatology, 1997

Background/Aims: Chronic hepatitis C develops in more than 70% of hepatitis C virus infected subj... more Background/Aims: Chronic hepatitis C develops in more than 70% of hepatitis C virus infected subjects. Viral factors influence the disease course, but little is known about the importance of host factors. Methods: Frequencies of major histocompatibility complex (MHC) class I and class II antigens were analyzed in two groups of patients with chronic hepatitis C virus infection and in control subjects. MHC class I typing was done by standard microlymphocytotoxicity assays. DRBl and DQAl genotyping was done by PCR based typing methods. Results: DRB1*0301 was found in 26 of 75 patients with chronic hepatitis C virus infection (34.7%) and in 12 of 101 control subjects (11.9%) (relative risk 3.9; p<O.OOl). Homozygosity for this allele appeared to confer a stronger risk. In contrast, DRB1*1301 was detected in three subjects with persistent infection HRONIC C infection with the hepatitis C virus (HCV) is a major health problem worldwide. In the United States 3.5 million people are believed to suffer from the disease (1). Chronic liver disease occurs in half of the HCV infected subjects. In a considerable proportion of patients chronic hepatitis may eventually progress to cirrhosis and hepatocellular carcinoma (2-4). Attempts to explain the clinical expression and the behaviour of chronic hepatitis C by viral factors have shown the importance of the viral genotypes and the level of viraemia for the clinical presentation (5,6). However, there remain large inconsistencies, and it is very likely that the immune response of the host can modify disease outcome. Clearance of the virus in

Human Kupffer cells secrete IL-10 in response to lipopolysaccharide (LPS) challenge

Journal of Hepatology, 1995

Kupffer cells are involved in local immunoregulation in the liver by secretion of cytokines and d... more Kupffer cells are involved in local immunoregulation in the liver by secretion of cytokines and direct cellular contact. They are able to influence the function of other liver cells, i.e. sinusoidal endothelial cells, Ito cells and hepatocytes. The three known major functions of Kupffer cells are clearance of endotoxin from the portal circulation, release of soluble mediators and presentation of antigen. Human Kupffer cells were isolated by collagenase perfusion followed by centrifugal elutriation and analyzed for cytokine secretion after 3 days in culture. We found that freshly isolated human Kupffer cells secreted the anti-inflammatory cytokine interleukin-10 in response to stimulation with lipopolysaccharide. The release of interleukin-10 was maximal 12-24 h after lipopolysaccharide challenge. Furthermore, we could show that exogenous interleukin-10 downregulated the release of the proinflammatory cytokines interleukin-6 and tumor necrosis factor alpha by Kupffer cells after lipopolysaccharide stimulation. The release of interleukin-6 was maximal 24 h after stimulation and interleukin-10 inhibited interleukin-6 release after 6 h. Tumor necrosis factor alpha showed maximal secretion 6 h after stimulation and exogenous interleukin-10 also downregulated the tumor necrosis factor alpha release after 6 h. Kupffer cells are exposed physiologically to lipopolysaccharide present in portal venous blood. Given the known anti-inflammatory effect of interleukin-10, our findings of secretion of interleukin-10 by Kupffer cells in response to lipopolysaccharide and suppression of interleukin-6 and tumor necrosis factor alpha release by Kupffer cells in vitro through exogenous interleukin-10 suggest an important role for interleukin-10 in the regulation of the local immune response in the liver sinusoid.

Regulation of chemokine (MCP-1, IP-10, MIP-2) secretion in liver sinusoidal endothelial cell by endotoxin, IL-10 and IL-4

Immunology Letters, 1997

Liver sinusoidal endothelial cells prime naive T cells and induce a Th2-shift implication for induction of tolerance in the liver

Immunology Letters, 1997