- Email: [email protected]
440 Increased TGFβ1 secretion via down-regulating NKG2D killer activator receptor expression results in impaired natural killer cell activity in patients with chronic HCV hepatitis
POSTERS
S164 5O 46 4e m 36
p
•
T
3e 25
F
{ 20 15 "~ 10 5 0
: NKTTNF-a
lgKTIFN-y NKTILl0 Cyto]dne~tP-.NAe~,l~ressionbyNKTIHL
IiT
i NKTTGF-p
Conclusion: Thl cytokine profile was predominant in NKT and Th2 was predominant in T-reg IHL in end-stage HCV-related chronic liver disease, suggesting that these cells might be responsible for HCV-related liver injury and fibrosis.
~HCV E21CD81 INTERACTIONS ALTER MIGRATION OF DENDRITIC CELLS IN CHRONIC HEPATITIS C H. Zimmermann, J. Nattermann, M. von Lilienfeld-Toal, L. Leifeld, T. Sauerbruch, U. Spengler. For the Kompetenznetz Hepatitis'. Department
of Internal Medicine I, University of Bonn, Bonn, Germany B a c k g r o u n d : Defects in the priming of antiviral immune responses have been suggested to contribute to the failure of viral clearance in HCV infection. Correct function of dendritic cells (DC) includes recruitment of DC to sites of inflammation where they take up antigens which after subsequent trafficking to the lymph nodes is presented to T-cells. Here, we studied compartmentalization of DC in chronic hepatitis C as well as HCV-induced changes in DC migration. Methods: Liver samples from HCV(+) patients (n 15) and HCV( ) controls (n 15) were immunohistochemically stained using anti-BDCA1and anti-BDCA2, which specifically stain DC. Circulating DC from 20 HCV(+) and 12 HCV( ) subjects were flowcytometrically studied. Monocyte-derived DC (MODC) and isolated CD8+ T-cells were stimulated with HCV E2 protein. DC migration was analyzed using a nitrocellulose filter micro chamber system. Results: We found a significantly lower proportion of circulating BDCA-1 and BDCA-2 expressing cells in HCV-infected patients than in healthy subjects. However, there was a significantly enhanced number of BDCA1(+) and BDCA-2(+) DC in livers from HCV(+) patients as compared to healthy controls (p <0.001). Triggering of CD81 on CD8+ T-cells with HCV E2 resulted in release of RANTES, the natural ligand of CCR5. As circulating DC express CCR5, we tested whether this affects DC migration. Indeed, we found that markedly more DC were attracted by the supernatant of HCV-E2-stimulated CD8+ T cells. Furthermore, incubation of DC with this supernatant resulted in down-regulation of CCR5 expression. Finally, we found that cross-linking of CD81 on MODC dramatically impaired the migration towards CCL21, a chemokine importantly involved in DC trafficking to lymphoid tissues. This effect was neither mediated by dysregulated maturation/differentiation nor by altering surface expression of chemokine receptors and integrins. Conclusion: We found an altered compartmentalization of DC in chronic hepatitis C. We propose HCV E2-mediated RANTES release to be involved in intrahepatic accumulation of DC. More importantly, our data suggest that cross-linking of CD81 on intrahepatic DC dramatically reduces the migratory response to CCL21. We suggest that this leads to an inefficient recruitment of DC to lymphoid tissues and therewith impaired T cell priming in chronic hepatitis C.
HEPATITIS C VIRAEMIA INHIBITS DENDRITIC CELL FUNCTIONS AND THEIR ABILITY TO INDUCE ADAPTIVE IMMUNE R E S P O N S E TO HCV
I. Pachiadakis 1, S. Chokshi 1, H. Cooksley 1, C. Sarazzin 2, S. Zeuzem 2, N.V. Naoumov1. 1The UCL Institute of Hepatology, University College
London, UK," 2Medizinische Klinik und Poliklinilc Innere Medizin II, Universitiitskliniken des Saarlandes, Homburg/Saar, Germany B a c k g r o u n d : Chronic infection with hepatitis C virus (HCV) is associated with impaired antiviral T-cell reactivity, which facilitates viral persistence. Whether the inefficient adaptive immune response is due to T-cell exhaustion/tolerance and/or to poor T-cell activation from the dendritic cells (DC) is not well understood. Study design/Methods: To determine whether early clearance of HCV viraemia during antiviral therapy improves primarily DC or T-cell functions, we studied paired samples of peripheral blood mononuclear cells (PBMC) from 22 patients with chronic hepatitis C: baseline (BL) and treatment week 12 (TW12) of treatment with peg-interferon-alpha 2b plus ribavirin. At TW12 serum HCV RNA was undetectable (TaqMan PCR) in 17/22 patients (responders), while 5/22 remained viraemic (non-responders). Monocyte-derived DC and purified CD4+ T-cells were obtained using standard protocols and analysed using flow cytometry (expression of CDla, CD40, CD80, CD83, CD86 and HLA-DR); IFN-7 response to recombinant HCV antigens (Elispot assay), mixed lymphocyte reaction (MLR) and interleukin-12 production. Furthermore, DC/CD4+ T-cell reactivity were tested in cross-over experiments using DC from BL tested with autologous T-cells from BL or TW12. Conversely, DC from TW12 were tested with autologous CD4+ T-cells from BL or TW12. HCV core protein was quantitated in plasma and DC lysates (Ortho assay). Results: The frequency of IFN-~/producing T-cells at TW12 was significantly higher in comparison with baseline in Responders, particularly the reactivity to HCV core protein, while the changes were not significant in Non-responders. Similarly the proliferative response (MLR) improved significantly only in responders. HCV core protein was detectable in DC and intracellular concentration at TW12 showed an inverse correlation with the frequency of IFN-7 producing T-cells. In the cross-over experiments, DC from TW12 induced greater number of IFN-g producing cells both from T-cell BL and TW12, in comparison to DC at baseline with corresponding T-cells in the same patient (p =0.045). Conclusion: These results indicate that the enhanced T-cell reactivity after HCV viraemia clearance early on treatment is due to improved dendritic cell function. Vaccine immunotherapy would require clearance of HCV viraemia to improve dendritic cell functions in order to generate effective T-cell responses.
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INCREASED TGFIH SECRETION VIA DOWN-REGULATING NKG2D KILLER ACTIVATOR RECEPTOR EXPRESSION RESULTS IN IMPAIRED NATURAL KILLER CELL ACTIVITY IN PATIENTS WITH CHRONIC HCV HEPATITIS
G. Par 1, T. Berki 2, L. Palinkas 2, L. Szereday 3, M. Halasz 3, J. SzekeresBartho 3, A. Miseta 4, G. Hegedfis 5, Zs. Faust6, Gy. Mozsik 1, B. Hunyady 1, A. Par 1. 1First Department of Medicine, 2Department of Immunology and
Biotechnology, 3Department of Medical Microbiology and Immunology, 4Department of Laboratory Medicine, University of Pecs, Hungary," 5Department of Pathology, Baranya County Hospital, Hungary," 6Blood Transfusion Centre, P~cs, Hungary Background and Aims: Impaired natural killer (NK) cell activity has been proposed as a mechanism contributing to viral persistence in hepatitis C virus (HCV) infection. NKG2D as an important activator receptor of NK cells, has a pivotal position in both innate and adaptive immunity since it is expressed not only on NK cells, but also on CD8+ T cells. Recent studies demonstrated that TGF[fil secreted by different tumors is responsible for
05C. Viral Hepatitis'
(e) Hepatitis' C
poor NK lytic activity via down-regulating NKG2D. Earlier in HCV hepatitis increased plasma TGF[Jl levels has been also demonstrated, now we analyzed whether decreased NK activity corresponds to a dysregulated expression of NKG2D. As regulatory T (Treg) cells are an important source of TGF[Jl, correlation between the percentage of Treg cells, plasma TGF[Jl levels and NKG2D expression of NK and T cells was also studied. Methods: The percentages of peripheral CD4+CD25high+Treg cells, NKG2D+ NK and T cells were determined by flow cytometry in 43 patients with chronic hepatitis C, in 10 sustained virological responders previously treated with IFN and in 15 healthy controls. Plasma levels of TGF[Jl were measured by ELISA. Results: In patients with chronic HCV hepatitis the activating receptor NKG2D expression was significantly (p < 0.001) downregulated both on NK (7.9• vs. 20.9• and on T cells (18• vs. 26.3• compared to healthy controls. This impaired expression of NKG2D was associated with increased proportion of CD4+CD25high+ Treg cells (4.6• vs.3.1• and increased TGF[Jl levels (15• vs. 9• compared to control group. TGF[Jl levels inversely correlated with the surface expression of NKG2D on NK cells (r 0.566). In contrast, the percentage of Treg cells (1.7• TGF[Jl levels ( l l . 6 • and the expression of NKG2D (NK: 17.1• T: 20.9• in sustained virological responders was comparable to that of healthy controls. Conclusion: Our data present the first evidence that TGF[Jl secreted by regulatory T cells is responsible for impaired NK cell function via down-regulating NKG2D activating receptor in chronic HCV hepatitis. Thus, TGF[J antagonism or soluble NKG2D ligands may provide the basis of a novel antiviral or even cancer immunotherapy to improve the function of NK and T cells.
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INFLUENCE OF INTERLEUKIN- 10 P R O M O T E R GENE POLYMORPHISM ON THE O C C U R R E N C E OF NON HODGKIN LYMPHOMA (NHL) IN SUBJECTS INFECTED WITH HEPATITIS C VIRUS (HCV)
M. Persico 1, M. Capasso 2, E. Persico 1, V. La Mura 1, M. Masarone 1, M. Svelto 1, A. Iolascon2. 1Internal Medicine and Hepatology Unit,
Second Universitiy of Naples, Italy," 2Biochemistry and Medical bioteehnologies departement, CEINGE Advanced Bioteehnologies, Federico H University of Naples, Italy HCV along with chronic liver disease is also considered a causative agent of other clinical pathological conditions which testify the possible direct pathogenic role of the virus in several different cell types including hepatocytes and leukocytes. Prevalence of HCV is significantly higher also in patients suffering with NHL and, all around the world, it was confirmed except for patients studied in North Europe and some areas of North America. In Italy, different groups showed prevalence ranging from 15 to 30%. The goal of this paper is to establish if a polymorphic gene encoding for cytokine could be a predisposing factor for this condition. To do this, we analyzed the distribution of the polymorphism of IL-10 1082 G/A in 126 patients, not infected with HCV, with Non Hodgkin Lymphoma (NHL/HCV-), in 100 patients, infected with HCV, with Non Hodgkin Lymphoma, (NHL/HCV+) and compared them to 80 patients with HCV related chronic hepatitis (CH/HCV) and 109 age, sex matched healthy blood donors. In this study, for the first time we show that irregardless of age, sex, virus genotype and/ or severity of chronic liver disease a significant prevalence of IL-10 1082 GG genoptype seems to influence the occurrence of NHL in HCV infected patients. In fact the distribution of the IL-10 1082 G/A polymorphism was different between NHL/HCV+ and NHL/HCV- patients (P 0.028), compared with GG homozygotes, the ORs of NHL/HCV+ patients for AG heterozygotes and AA homozygotes were 0.33 (95% confidence interval, 0.14 0.83) and 0.32 (95% confidence interval, 0.12 0.89) respectively. The frequency of the IL-10 1082 G allele (P 0.019) and the frequency of the IL-10 -1082 GG genotype
Experimental (immunology)
$165
against overall genotypes (IL-10 1082 GA/AA) were significantly higher in NHL/HCV+ patients as compared with NHL/HCV- patients (P 0.014). In conclusion the interleukin-10 promoter gene polymorphism influences the occurrence of NHL in subjects infected with HCV.
[•
TNF-ALPHA AND IL-10 GENETIC POLYMORPHISMS INFLUENCE THE NATURAL HISTORY AND THE RESPONSE TO ANTIVIRAL T H E R A P Y IN HCV RELATED CHRONIC HEPATITIS
M. Persico 1, M. Capasso 2, E. Persico 1, M. Svelto 1, R. Russo 2, M. Masarone 1, A. Iolascon 2. 1Internal Medicine and Hepatology
Unit, Second University of Naples, Italy, 2Biochemistry and Medical bioteehnologies departement, CEINGE Advanced Biotechnologies, Federico H University of Naples, Italy Chronic hepatitis C (CH-C) develops in 33 84% of patients with acute hepatitis. About one-third of patients develop liver cirrhosis within 15 25 years with an annual risk of hepatocellular carcinoma in 1 4%. HCV positive patients with persistent normal ALT (PNAL) represent the 20 30% of the HCV population and the natural history of the disease in these subjects is benign. Antiviral therapy of CHC yields HCV eradication rates from 54% to 56%. Here, we verify the potential role of cytokine in inflammatory damage of CHC and in IFN therapy resistance. So, polymorphisms in IL-10 and TNF-a ( 1082 G/A and 308 G/A, respectively) were analyzed in: 156 CHC subjects; 45 with cirrhosis; 34 CHC responder t (CHC R) and 46 CHC non-responder (CHC NR); 22 PNAL patients, TNFa308 GG homozygous individuals were classified as "low-TNF producers"; TNFa308 A carrier individuals were classified as "high-TNF producers"; IL-10 1082 A carrier individuals were classified as "low-IL-10 producers"; IL-10 1082 GG homozygous individuals were classified as high-IL-10 producers". A significant decrease of the lowTNF-alpha?/high-IL-10 producer genotype vs all the other genotypes was observed in Cirrhotic group compared with CHC subjects (P 0.011). This is supported by studies demonstrating that IL-10 reduces necroinflammatory activity and progression of fibrosis in patients with chronic hepatitis. In CHC NR patients the high-TNF/high-IL-10 producer genotype versus the other genotypes increased significantly as compared with CHC R patients (P 0.048). This seems in agreement with the demonstration that increased IL-10 and TNF-a production is associated with a poor response to INE Finally, the arranged genotype distribution did not significantly differ between PNAL and CHC groups. Instead, the frequency of TNFA allele (high producer) is significantly increased (P 0.048) in PNAL subjects. Our results suggest that: 1. PNAL patients seems characterized by TNF-a high producer. 2. The TNF-alpha high producer/IL-101ow producer seems characterizing cirrothic patients. 3. The TNF-alpha high producer/IL-10high producer seems influencing the negative response to antiviral treatment.
[•
HEPATITIS C VIRUS PROMOTES ANGIOPOIETIN-2 EXPRESSION IN LIVER TISSUE OF PATIENTS WITH CHRONIC HEPATITIS C
P. Sanz-Camenol., S. Mart{n-Vilchez 1, M.J. Borque 2, J.A. MorenoMonteagudo 1, L. Garcla-Buey 1, M. Ldpez-Cabrera 2, R. Moreno-Otero 1.
1Liver Unit, 2Molecular Biology Unit/Hospital Universitario de la Princesa, Madrid, Spain Background: Previous studies performed by our group described the intrahepatic expression of iNOS (Majano, JCI 1998), nitrotyrosine accumulation (Sanz-Cameno, J. Hepatol. 2002) and VEGF synthesis (Medina, J Hepatol 2003) in patients with chronic liver disease, molecules directly implicated in neoangiogenic events. We have also observed in primary billiary cirrhosis the expression of angiopoietin-2 (Ang-2) (Sanz-Cameno, J. Hepatol. 2005), a relevant protein in vascular development, integrity
S164 5O 46 4e m 36
p
•
T
3e 25
F
{ 20 15 "~ 10 5 0
: NKTTNF-a
lgKTIFN-y NKTILl0 Cyto]dne~tP-.NAe~,l~ressionbyNKTIHL
IiT
i NKTTGF-p
Conclusion: Thl cytokine profile was predominant in NKT and Th2 was predominant in T-reg IHL in end-stage HCV-related chronic liver disease, suggesting that these cells might be responsible for HCV-related liver injury and fibrosis.
~HCV E21CD81 INTERACTIONS ALTER MIGRATION OF DENDRITIC CELLS IN CHRONIC HEPATITIS C H. Zimmermann, J. Nattermann, M. von Lilienfeld-Toal, L. Leifeld, T. Sauerbruch, U. Spengler. For the Kompetenznetz Hepatitis'. Department
of Internal Medicine I, University of Bonn, Bonn, Germany B a c k g r o u n d : Defects in the priming of antiviral immune responses have been suggested to contribute to the failure of viral clearance in HCV infection. Correct function of dendritic cells (DC) includes recruitment of DC to sites of inflammation where they take up antigens which after subsequent trafficking to the lymph nodes is presented to T-cells. Here, we studied compartmentalization of DC in chronic hepatitis C as well as HCV-induced changes in DC migration. Methods: Liver samples from HCV(+) patients (n 15) and HCV( ) controls (n 15) were immunohistochemically stained using anti-BDCA1and anti-BDCA2, which specifically stain DC. Circulating DC from 20 HCV(+) and 12 HCV( ) subjects were flowcytometrically studied. Monocyte-derived DC (MODC) and isolated CD8+ T-cells were stimulated with HCV E2 protein. DC migration was analyzed using a nitrocellulose filter micro chamber system. Results: We found a significantly lower proportion of circulating BDCA-1 and BDCA-2 expressing cells in HCV-infected patients than in healthy subjects. However, there was a significantly enhanced number of BDCA1(+) and BDCA-2(+) DC in livers from HCV(+) patients as compared to healthy controls (p <0.001). Triggering of CD81 on CD8+ T-cells with HCV E2 resulted in release of RANTES, the natural ligand of CCR5. As circulating DC express CCR5, we tested whether this affects DC migration. Indeed, we found that markedly more DC were attracted by the supernatant of HCV-E2-stimulated CD8+ T cells. Furthermore, incubation of DC with this supernatant resulted in down-regulation of CCR5 expression. Finally, we found that cross-linking of CD81 on MODC dramatically impaired the migration towards CCL21, a chemokine importantly involved in DC trafficking to lymphoid tissues. This effect was neither mediated by dysregulated maturation/differentiation nor by altering surface expression of chemokine receptors and integrins. Conclusion: We found an altered compartmentalization of DC in chronic hepatitis C. We propose HCV E2-mediated RANTES release to be involved in intrahepatic accumulation of DC. More importantly, our data suggest that cross-linking of CD81 on intrahepatic DC dramatically reduces the migratory response to CCL21. We suggest that this leads to an inefficient recruitment of DC to lymphoid tissues and therewith impaired T cell priming in chronic hepatitis C.
HEPATITIS C VIRAEMIA INHIBITS DENDRITIC CELL FUNCTIONS AND THEIR ABILITY TO INDUCE ADAPTIVE IMMUNE R E S P O N S E TO HCV
I. Pachiadakis 1, S. Chokshi 1, H. Cooksley 1, C. Sarazzin 2, S. Zeuzem 2, N.V. Naoumov1. 1The UCL Institute of Hepatology, University College
London, UK," 2Medizinische Klinik und Poliklinilc Innere Medizin II, Universitiitskliniken des Saarlandes, Homburg/Saar, Germany B a c k g r o u n d : Chronic infection with hepatitis C virus (HCV) is associated with impaired antiviral T-cell reactivity, which facilitates viral persistence. Whether the inefficient adaptive immune response is due to T-cell exhaustion/tolerance and/or to poor T-cell activation from the dendritic cells (DC) is not well understood. Study design/Methods: To determine whether early clearance of HCV viraemia during antiviral therapy improves primarily DC or T-cell functions, we studied paired samples of peripheral blood mononuclear cells (PBMC) from 22 patients with chronic hepatitis C: baseline (BL) and treatment week 12 (TW12) of treatment with peg-interferon-alpha 2b plus ribavirin. At TW12 serum HCV RNA was undetectable (TaqMan PCR) in 17/22 patients (responders), while 5/22 remained viraemic (non-responders). Monocyte-derived DC and purified CD4+ T-cells were obtained using standard protocols and analysed using flow cytometry (expression of CDla, CD40, CD80, CD83, CD86 and HLA-DR); IFN-7 response to recombinant HCV antigens (Elispot assay), mixed lymphocyte reaction (MLR) and interleukin-12 production. Furthermore, DC/CD4+ T-cell reactivity were tested in cross-over experiments using DC from BL tested with autologous T-cells from BL or TW12. Conversely, DC from TW12 were tested with autologous CD4+ T-cells from BL or TW12. HCV core protein was quantitated in plasma and DC lysates (Ortho assay). Results: The frequency of IFN-~/producing T-cells at TW12 was significantly higher in comparison with baseline in Responders, particularly the reactivity to HCV core protein, while the changes were not significant in Non-responders. Similarly the proliferative response (MLR) improved significantly only in responders. HCV core protein was detectable in DC and intracellular concentration at TW12 showed an inverse correlation with the frequency of IFN-7 producing T-cells. In the cross-over experiments, DC from TW12 induced greater number of IFN-g producing cells both from T-cell BL and TW12, in comparison to DC at baseline with corresponding T-cells in the same patient (p =0.045). Conclusion: These results indicate that the enhanced T-cell reactivity after HCV viraemia clearance early on treatment is due to improved dendritic cell function. Vaccine immunotherapy would require clearance of HCV viraemia to improve dendritic cell functions in order to generate effective T-cell responses.
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INCREASED TGFIH SECRETION VIA DOWN-REGULATING NKG2D KILLER ACTIVATOR RECEPTOR EXPRESSION RESULTS IN IMPAIRED NATURAL KILLER CELL ACTIVITY IN PATIENTS WITH CHRONIC HCV HEPATITIS
G. Par 1, T. Berki 2, L. Palinkas 2, L. Szereday 3, M. Halasz 3, J. SzekeresBartho 3, A. Miseta 4, G. Hegedfis 5, Zs. Faust6, Gy. Mozsik 1, B. Hunyady 1, A. Par 1. 1First Department of Medicine, 2Department of Immunology and
Biotechnology, 3Department of Medical Microbiology and Immunology, 4Department of Laboratory Medicine, University of Pecs, Hungary," 5Department of Pathology, Baranya County Hospital, Hungary," 6Blood Transfusion Centre, P~cs, Hungary Background and Aims: Impaired natural killer (NK) cell activity has been proposed as a mechanism contributing to viral persistence in hepatitis C virus (HCV) infection. NKG2D as an important activator receptor of NK cells, has a pivotal position in both innate and adaptive immunity since it is expressed not only on NK cells, but also on CD8+ T cells. Recent studies demonstrated that TGF[fil secreted by different tumors is responsible for
05C. Viral Hepatitis'
(e) Hepatitis' C
poor NK lytic activity via down-regulating NKG2D. Earlier in HCV hepatitis increased plasma TGF[Jl levels has been also demonstrated, now we analyzed whether decreased NK activity corresponds to a dysregulated expression of NKG2D. As regulatory T (Treg) cells are an important source of TGF[Jl, correlation between the percentage of Treg cells, plasma TGF[Jl levels and NKG2D expression of NK and T cells was also studied. Methods: The percentages of peripheral CD4+CD25high+Treg cells, NKG2D+ NK and T cells were determined by flow cytometry in 43 patients with chronic hepatitis C, in 10 sustained virological responders previously treated with IFN and in 15 healthy controls. Plasma levels of TGF[Jl were measured by ELISA. Results: In patients with chronic HCV hepatitis the activating receptor NKG2D expression was significantly (p < 0.001) downregulated both on NK (7.9• vs. 20.9• and on T cells (18• vs. 26.3• compared to healthy controls. This impaired expression of NKG2D was associated with increased proportion of CD4+CD25high+ Treg cells (4.6• vs.3.1• and increased TGF[Jl levels (15• vs. 9• compared to control group. TGF[Jl levels inversely correlated with the surface expression of NKG2D on NK cells (r 0.566). In contrast, the percentage of Treg cells (1.7• TGF[Jl levels ( l l . 6 • and the expression of NKG2D (NK: 17.1• T: 20.9• in sustained virological responders was comparable to that of healthy controls. Conclusion: Our data present the first evidence that TGF[Jl secreted by regulatory T cells is responsible for impaired NK cell function via down-regulating NKG2D activating receptor in chronic HCV hepatitis. Thus, TGF[J antagonism or soluble NKG2D ligands may provide the basis of a novel antiviral or even cancer immunotherapy to improve the function of NK and T cells.
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INFLUENCE OF INTERLEUKIN- 10 P R O M O T E R GENE POLYMORPHISM ON THE O C C U R R E N C E OF NON HODGKIN LYMPHOMA (NHL) IN SUBJECTS INFECTED WITH HEPATITIS C VIRUS (HCV)
M. Persico 1, M. Capasso 2, E. Persico 1, V. La Mura 1, M. Masarone 1, M. Svelto 1, A. Iolascon2. 1Internal Medicine and Hepatology Unit,
Second Universitiy of Naples, Italy," 2Biochemistry and Medical bioteehnologies departement, CEINGE Advanced Bioteehnologies, Federico H University of Naples, Italy HCV along with chronic liver disease is also considered a causative agent of other clinical pathological conditions which testify the possible direct pathogenic role of the virus in several different cell types including hepatocytes and leukocytes. Prevalence of HCV is significantly higher also in patients suffering with NHL and, all around the world, it was confirmed except for patients studied in North Europe and some areas of North America. In Italy, different groups showed prevalence ranging from 15 to 30%. The goal of this paper is to establish if a polymorphic gene encoding for cytokine could be a predisposing factor for this condition. To do this, we analyzed the distribution of the polymorphism of IL-10 1082 G/A in 126 patients, not infected with HCV, with Non Hodgkin Lymphoma (NHL/HCV-), in 100 patients, infected with HCV, with Non Hodgkin Lymphoma, (NHL/HCV+) and compared them to 80 patients with HCV related chronic hepatitis (CH/HCV) and 109 age, sex matched healthy blood donors. In this study, for the first time we show that irregardless of age, sex, virus genotype and/ or severity of chronic liver disease a significant prevalence of IL-10 1082 GG genoptype seems to influence the occurrence of NHL in HCV infected patients. In fact the distribution of the IL-10 1082 G/A polymorphism was different between NHL/HCV+ and NHL/HCV- patients (P 0.028), compared with GG homozygotes, the ORs of NHL/HCV+ patients for AG heterozygotes and AA homozygotes were 0.33 (95% confidence interval, 0.14 0.83) and 0.32 (95% confidence interval, 0.12 0.89) respectively. The frequency of the IL-10 1082 G allele (P 0.019) and the frequency of the IL-10 -1082 GG genotype
Experimental (immunology)
$165
against overall genotypes (IL-10 1082 GA/AA) were significantly higher in NHL/HCV+ patients as compared with NHL/HCV- patients (P 0.014). In conclusion the interleukin-10 promoter gene polymorphism influences the occurrence of NHL in subjects infected with HCV.
[•
TNF-ALPHA AND IL-10 GENETIC POLYMORPHISMS INFLUENCE THE NATURAL HISTORY AND THE RESPONSE TO ANTIVIRAL T H E R A P Y IN HCV RELATED CHRONIC HEPATITIS
M. Persico 1, M. Capasso 2, E. Persico 1, M. Svelto 1, R. Russo 2, M. Masarone 1, A. Iolascon 2. 1Internal Medicine and Hepatology
Unit, Second University of Naples, Italy, 2Biochemistry and Medical bioteehnologies departement, CEINGE Advanced Biotechnologies, Federico H University of Naples, Italy Chronic hepatitis C (CH-C) develops in 33 84% of patients with acute hepatitis. About one-third of patients develop liver cirrhosis within 15 25 years with an annual risk of hepatocellular carcinoma in 1 4%. HCV positive patients with persistent normal ALT (PNAL) represent the 20 30% of the HCV population and the natural history of the disease in these subjects is benign. Antiviral therapy of CHC yields HCV eradication rates from 54% to 56%. Here, we verify the potential role of cytokine in inflammatory damage of CHC and in IFN therapy resistance. So, polymorphisms in IL-10 and TNF-a ( 1082 G/A and 308 G/A, respectively) were analyzed in: 156 CHC subjects; 45 with cirrhosis; 34 CHC responder t (CHC R) and 46 CHC non-responder (CHC NR); 22 PNAL patients, TNFa308 GG homozygous individuals were classified as "low-TNF producers"; TNFa308 A carrier individuals were classified as "high-TNF producers"; IL-10 1082 A carrier individuals were classified as "low-IL-10 producers"; IL-10 1082 GG homozygous individuals were classified as high-IL-10 producers". A significant decrease of the lowTNF-alpha?/high-IL-10 producer genotype vs all the other genotypes was observed in Cirrhotic group compared with CHC subjects (P 0.011). This is supported by studies demonstrating that IL-10 reduces necroinflammatory activity and progression of fibrosis in patients with chronic hepatitis. In CHC NR patients the high-TNF/high-IL-10 producer genotype versus the other genotypes increased significantly as compared with CHC R patients (P 0.048). This seems in agreement with the demonstration that increased IL-10 and TNF-a production is associated with a poor response to INE Finally, the arranged genotype distribution did not significantly differ between PNAL and CHC groups. Instead, the frequency of TNFA allele (high producer) is significantly increased (P 0.048) in PNAL subjects. Our results suggest that: 1. PNAL patients seems characterized by TNF-a high producer. 2. The TNF-alpha high producer/IL-101ow producer seems characterizing cirrothic patients. 3. The TNF-alpha high producer/IL-10high producer seems influencing the negative response to antiviral treatment.
[•
HEPATITIS C VIRUS PROMOTES ANGIOPOIETIN-2 EXPRESSION IN LIVER TISSUE OF PATIENTS WITH CHRONIC HEPATITIS C
P. Sanz-Camenol., S. Mart{n-Vilchez 1, M.J. Borque 2, J.A. MorenoMonteagudo 1, L. Garcla-Buey 1, M. Ldpez-Cabrera 2, R. Moreno-Otero 1.
1Liver Unit, 2Molecular Biology Unit/Hospital Universitario de la Princesa, Madrid, Spain Background: Previous studies performed by our group described the intrahepatic expression of iNOS (Majano, JCI 1998), nitrotyrosine accumulation (Sanz-Cameno, J. Hepatol. 2002) and VEGF synthesis (Medina, J Hepatol 2003) in patients with chronic liver disease, molecules directly implicated in neoangiogenic events. We have also observed in primary billiary cirrhosis the expression of angiopoietin-2 (Ang-2) (Sanz-Cameno, J. Hepatol. 2005), a relevant protein in vascular development, integrity