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631 Endotoxin receptor CD14 and TLR4 polymorphisms in chronic liver disease
Category 6: Innate and Adaptive Immunity of the Liver (+1335) and 186,000/mm 3 (+60.9) respectivdy. The mean time of followup was 8(+3.5) months. Nineteen patients showed a decrease in the Hb levels, 32 a decrease in neutrophil count, and 10 a decrease in platelet count. A BL Hb level below 14g]dl (OR: 4.9, 95%CI: 1.7-13.8) and treatment with AZT (OR: 4.2, 95%CI: 1.5-12.1) were the independent factors assodated with a higher risk for the appearance of a Hb level <10~dl during the follow-up. For nentropenia three independent factors were found: BL neutrophil count <2910/mm 3 (OR: 2.8, 95%CI: 1.2-6.4), weight <60kg (OR: 3.3, 95%CI: 1.3-7.9) and female gender (OR: 2.9, 95%CI: 1.2-6.8). For plaquetopenia only a BL platelet count (OR: 0.9, 95%CI: 0.85-0.97) was associated with a higher risk of a platelet count <50,000/ram 3 . Conclusions: Special care should be taken when prescribing anti-HCV therapy for a subset of coinfected patients because they might be at higher risk of suffering peginterferon o~-2a or ribavirin dosage reductions. Strategies to improve adherence in these patients should be actively searched.
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ENDOTOXIN RECEPTOR CD14 AND TLR4 POLYMORPHISMS IN CHRONIC LIVER DISEASE
J. Halangk 1, T. von Hahn 1, H. Witt 2, G. Puhl 3, T. M~tller1, B. Wiedenmann I , R Neuhaus 3, U. Beners 4, W. Luck 2, R. Nickel 2,
T. BergI . IHepatology, Gastroenterology, CT~aritd Campus VirchowKlinikum, Berlin, Germany," 2pediatric's, Charitd Campus VirchowKlinikum, Berlin, Germany,"3Surgery, Charit~ Campus Virchow-Kl#aikum Berlin, Germany," 4 Gastroenterolog)~. Klinikum Grosshadern, Munich, Germany Introduction: Activation of Kupffer cells by lipopolysaccharides (LPS) is mediated through the CD14/TLR4 receptor complex. The ensuing release of inflammatory cytoldnes such as tumor necrosis factor oc (TNF-~) by Kupffer cells prmnotes liver injury. A 260C>T transition in the CD14 prmnoter has been reported to increase the expression ofCD14 resnlting in enhanced LPS responsiveness and a more progressive course of alcoholic liver disease. On the other hand, a D299G exchange in the TLR4 gene diminishes the fifftammatory response to LPS but its relevance in chronic liver disease has not been evaluated yet. Thus, we investigated the role of these two genetic alterations in the development and progression of chrmfic liver diseases. Patients and Methods: We performed genotyping by PCR amplification and melting curve analysis with FRET probes in 1656 patients with chronic liver disease of different etiologies (i.e. viral: n=851, antoimmune: n 411, alcoholic: n 227, cryptogenic: n 65, persistently elevated transaminases: n=29, other: n= 73) and in 385 healthy controls. Of these patients, 737 (44.5%) underwent orthotopic liver transplantation (OLT) because of decmnpensated liver disease. Results: Genotype frequencies o f CD14 260C>T and TLR4 D299G did not significantly differ between patients and controls (CD14 TT 21.7% vs. 21.8%; TLR4 DG or GG 9.7% vs. 10.4%). Furthermore, differences in genotype frequencies were neither found between patients with compensated (no OLT) and decompensated (OLT group) disease nor between those with different etiologies. However, in a subgroup of patients (n 29) with persistently elevated transaminases without any hint for the etiology of the liver disorder, a significantly higher frequency of the CD14 -260TT genotype was observed (37.9% vs. 21.4% for other diagnoses, p = 0.03). Conclusion: This large scale study does not reveal an impact of the investigated CD14 and TLR4 polymorphisms on the incidence and outcome o f chronic liver diseases. The finding of a significantly increased frequency of homozygotes for the CD14 -260C>T promoter polymorphism in patients with persistently elevated transaminases in the absence of a readily identifiable etiology is interesting and warrants further investigation. Hepatic toxicity of microbial components mediated via increased responsiveness of the CD14/TLR4 pathway in otherwise healthy individuals might be a favorable hypothesis.
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231
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MANNOSE-BINDING LECTIN (MBL2) POLYMORPHISMS IN PATIENTS WITH CHRONIC LIVER DISEASE OF DIFFERENT ETIOLOGIES
J. Halangk 1, H. Witt 2, A. Hachfeld1, G. Puhl 3, T. MiillerI , R. Nickel 2, B. Wiedenmann I , P. Neuhaus 3, W. Luck 2, T. Berg I . IHepatology,
Gastroenterology, Char#d, Campus IOrchow-Klinikum,Berlin, Germany; 2Pediatrics, CharitY, Campus Virchow-Klinikum, Berlin, Germany; eSurgery, CharitY, Campus Virchow-Klinikum, Berlin, Germany Introduction: The mannose-binding lectin (MBL2) is a plasma collectin exerting innate immune defence functions like opsonisation and complement activation. MBL plasma concentration is mainly determined by a series of allelic variants located in exon 1 (G54D, G57E, R52C, designated B, C, D, respectively), in the promoter region (H/L and X/Y) and in the 5e untranslated region (P/Q). Individuals homozygous for the exon 1 polymorphisms produce virtually undetectable MBL levels whereas heterozygotes have markedly diminished MBL levels (about 10%) in comparison to wildtype homozygotes. MBL2 polymorphisms have been associated with a variety of infectious and autoimnmne diseases. Several studies in patients with viral and autoimmune liver disease have brought conflicting results. The afin of our study was to investigate the sig-nificance ofMBL2 exon 1 (B, D, C), promoter (X/Y) and 5'UTR (P/Q) polymorphisms in chronic liver disease of multiple etiologies in a large population of 1539 patients and 830 healthy controls. Patients and Methods: We included 225 patients with alcoholic liver disease, 40 patients with acute liver failure, 651 patients with HCV, 203 patients with HBV, 98 patients with autoimmune hepatitis, 209 patients with PBC or PSC, 65 patients with cryptogenic cirrhosis and 48 patients with NAFLD. A birth cohort of healthy newborns served as a control population. Genotyping was performed by PCR amplification and melting curve analysis with FRET probes. Results: We found no differences in genotype frequencies of the MBL2 polyrnorphisms between patients and controls. Genotype and allele frequencies were comparable to those reported in previous studied European populations. There was no overrepresentation of the MBL2 polymorphisms in any of the groups of patients with different liver diseases investigated. MBL2 polymorphisms also neither influenced histologic grade of fidtammation nor stage of fibrosis as documented mainly in patients with chronic hepatitis C. Furthermore, between patients with (n=809) and without cirrhosis (n 730) no difference in the MBL2 genotype distribution could be observed. Conclusion: This large genetic association study could exclude MBL2 polymorphisms as major genetic risk factors for the development and progression of chronic liver disease.
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HEPATIC CD40L OVEREXPRESSION INDUCES FULMINANT HEPATITIS IN MICE, ESTABLISHMENT OF A NEW MODEL OF IMMUNOMEDIATED FULMINANT HEPATIC FAILURE
L. Leifeld1, F. Dombrowski 2, J. Prieto ~, C. Qian ~, T. Sauerbruch 1, W. Caselmann 1, U. Spengler I , V Schmitz I . 1Department of Internal
Medicine I - University of Bonn, Bonn, Germany; 2Institute of Pathology University of Magdebu~, Magdebu~, Germany," 3Gone Therapy Unit, Universidad de Navarra, Pamplona, Spain A dysbalance in immune-activation is discussed as a major pathophysiological mechanism for the development of fulminant hepatic failure (FHF). In previous studies in hmnan and murine FHF we demonstrated a strong intrahepatic upregulation of tile costimnlatory molecules CD40 and CD40 ligand (Am. J. Pathol. 1999; 154). Now, we established a new model of fulminant hepatitis by induction of hepatic overexpression of CD40L by adenoviral gone transfer in mice (AdCD40L, control vector: AdlacZ). Fnlminant hepatic failure was characterized by steadily increasing periportal
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ENDOTOXIN RECEPTOR CD14 AND TLR4 POLYMORPHISMS IN CHRONIC LIVER DISEASE
J. Halangk 1, T. von Hahn 1, H. Witt 2, G. Puhl 3, T. M~tller1, B. Wiedenmann I , R Neuhaus 3, U. Beners 4, W. Luck 2, R. Nickel 2,
T. BergI . IHepatology, Gastroenterology, CT~aritd Campus VirchowKlinikum, Berlin, Germany," 2pediatric's, Charitd Campus VirchowKlinikum, Berlin, Germany,"3Surgery, Charit~ Campus Virchow-Kl#aikum Berlin, Germany," 4 Gastroenterolog)~. Klinikum Grosshadern, Munich, Germany Introduction: Activation of Kupffer cells by lipopolysaccharides (LPS) is mediated through the CD14/TLR4 receptor complex. The ensuing release of inflammatory cytoldnes such as tumor necrosis factor oc (TNF-~) by Kupffer cells prmnotes liver injury. A 260C>T transition in the CD14 prmnoter has been reported to increase the expression ofCD14 resnlting in enhanced LPS responsiveness and a more progressive course of alcoholic liver disease. On the other hand, a D299G exchange in the TLR4 gene diminishes the fifftammatory response to LPS but its relevance in chronic liver disease has not been evaluated yet. Thus, we investigated the role of these two genetic alterations in the development and progression of chrmfic liver diseases. Patients and Methods: We performed genotyping by PCR amplification and melting curve analysis with FRET probes in 1656 patients with chronic liver disease of different etiologies (i.e. viral: n=851, antoimmune: n 411, alcoholic: n 227, cryptogenic: n 65, persistently elevated transaminases: n=29, other: n= 73) and in 385 healthy controls. Of these patients, 737 (44.5%) underwent orthotopic liver transplantation (OLT) because of decmnpensated liver disease. Results: Genotype frequencies o f CD14 260C>T and TLR4 D299G did not significantly differ between patients and controls (CD14 TT 21.7% vs. 21.8%; TLR4 DG or GG 9.7% vs. 10.4%). Furthermore, differences in genotype frequencies were neither found between patients with compensated (no OLT) and decompensated (OLT group) disease nor between those with different etiologies. However, in a subgroup of patients (n 29) with persistently elevated transaminases without any hint for the etiology of the liver disorder, a significantly higher frequency of the CD14 -260TT genotype was observed (37.9% vs. 21.4% for other diagnoses, p = 0.03). Conclusion: This large scale study does not reveal an impact of the investigated CD14 and TLR4 polymorphisms on the incidence and outcome o f chronic liver diseases. The finding of a significantly increased frequency of homozygotes for the CD14 -260C>T promoter polymorphism in patients with persistently elevated transaminases in the absence of a readily identifiable etiology is interesting and warrants further investigation. Hepatic toxicity of microbial components mediated via increased responsiveness of the CD14/TLR4 pathway in otherwise healthy individuals might be a favorable hypothesis.
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231
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MANNOSE-BINDING LECTIN (MBL2) POLYMORPHISMS IN PATIENTS WITH CHRONIC LIVER DISEASE OF DIFFERENT ETIOLOGIES
J. Halangk 1, H. Witt 2, A. Hachfeld1, G. Puhl 3, T. MiillerI , R. Nickel 2, B. Wiedenmann I , P. Neuhaus 3, W. Luck 2, T. Berg I . IHepatology,
Gastroenterology, Char#d, Campus IOrchow-Klinikum,Berlin, Germany; 2Pediatrics, CharitY, Campus Virchow-Klinikum, Berlin, Germany; eSurgery, CharitY, Campus Virchow-Klinikum, Berlin, Germany Introduction: The mannose-binding lectin (MBL2) is a plasma collectin exerting innate immune defence functions like opsonisation and complement activation. MBL plasma concentration is mainly determined by a series of allelic variants located in exon 1 (G54D, G57E, R52C, designated B, C, D, respectively), in the promoter region (H/L and X/Y) and in the 5e untranslated region (P/Q). Individuals homozygous for the exon 1 polymorphisms produce virtually undetectable MBL levels whereas heterozygotes have markedly diminished MBL levels (about 10%) in comparison to wildtype homozygotes. MBL2 polymorphisms have been associated with a variety of infectious and autoimnmne diseases. Several studies in patients with viral and autoimmune liver disease have brought conflicting results. The afin of our study was to investigate the sig-nificance ofMBL2 exon 1 (B, D, C), promoter (X/Y) and 5'UTR (P/Q) polymorphisms in chronic liver disease of multiple etiologies in a large population of 1539 patients and 830 healthy controls. Patients and Methods: We included 225 patients with alcoholic liver disease, 40 patients with acute liver failure, 651 patients with HCV, 203 patients with HBV, 98 patients with autoimmune hepatitis, 209 patients with PBC or PSC, 65 patients with cryptogenic cirrhosis and 48 patients with NAFLD. A birth cohort of healthy newborns served as a control population. Genotyping was performed by PCR amplification and melting curve analysis with FRET probes. Results: We found no differences in genotype frequencies of the MBL2 polyrnorphisms between patients and controls. Genotype and allele frequencies were comparable to those reported in previous studied European populations. There was no overrepresentation of the MBL2 polymorphisms in any of the groups of patients with different liver diseases investigated. MBL2 polymorphisms also neither influenced histologic grade of fidtammation nor stage of fibrosis as documented mainly in patients with chronic hepatitis C. Furthermore, between patients with (n=809) and without cirrhosis (n 730) no difference in the MBL2 genotype distribution could be observed. Conclusion: This large genetic association study could exclude MBL2 polymorphisms as major genetic risk factors for the development and progression of chronic liver disease.
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HEPATIC CD40L OVEREXPRESSION INDUCES FULMINANT HEPATITIS IN MICE, ESTABLISHMENT OF A NEW MODEL OF IMMUNOMEDIATED FULMINANT HEPATIC FAILURE
L. Leifeld1, F. Dombrowski 2, J. Prieto ~, C. Qian ~, T. Sauerbruch 1, W. Caselmann 1, U. Spengler I , V Schmitz I . 1Department of Internal
Medicine I - University of Bonn, Bonn, Germany; 2Institute of Pathology University of Magdebu~, Magdebu~, Germany," 3Gone Therapy Unit, Universidad de Navarra, Pamplona, Spain A dysbalance in immune-activation is discussed as a major pathophysiological mechanism for the development of fulminant hepatic failure (FHF). In previous studies in hmnan and murine FHF we demonstrated a strong intrahepatic upregulation of tile costimnlatory molecules CD40 and CD40 ligand (Am. J. Pathol. 1999; 154). Now, we established a new model of fulminant hepatitis by induction of hepatic overexpression of CD40L by adenoviral gone transfer in mice (AdCD40L, control vector: AdlacZ). Fnlminant hepatic failure was characterized by steadily increasing periportal