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Non-alcoholic steatohepatitis (NASH) and hereditary hemochromatosis (HHC)
A1264 AASLD ABSTRACTS
• L0271 SUPPRESSION OF PBC-LIKE HEPATIC LESIONS IN MURINE GVHR M O D E L BY ANTIBODIES AGAINST ADHESION MOLECULES. S. Itoh, Y. Matsuzaki, T. Kimura, T. Ikegami, J. Shoda, N. Tanaka Tsukuba, Ibaraki, Japan. Background: We have previously demonstrated that Thl type cytokine
mRNA firstly increased and Th2 type cytokine mRNA elevated subsequently in liver infiltrating CD4 ÷ T cells of PBC-like hepatic lesion using murine GVHR (Gastroenterology, 110, A1219, 1996). A number of cell adhesion molecules are important for CD4 ÷ T ceils to infiltrate the liver. Moreover, we have also shown that antibodies against adhesion molecules such as LFA-1 and VLA-4 suppressed the formation of PBC-like lesions (Hepatology, 22 (4) pt.2, 1995, Gastroenterology 110 (4) A1235, 1996). However, the precise molecular mechanism of these suppressions by the administration of antibodies against adhesion molecules still remains unknown. Aim: To clarify whether Thl/Th2 balance is related to the suppression of PBC-like hepatic lesions by antibodies against adhesion molecules. Methods: Sex-matched 1 x 107 C57BL/6 (B6) T spleen cells were injected into (B6.C-H-2 bml2 x B6) F 1 mice intravenously. Each 50 lag of anti-mouse t~ chain of VLA-4 mAb (PS2.3), anti-mouse VCAM-1 mAb (MK/2) or normal rat IgG were administered intraperitoneally per mouse during cell transfer. All mice were sacrificed 2 weeks after the first cell transfer. 1 x 10-s liver infiltrating Thy 1.2+CD4÷ lymphocytes were sorted using FACS vantage and the RNA was isolated by AGPC method. A semiquantitative RT-PCR was carried out using primers specific for IL-2, IFN~, IL-4, IL-10. Immunohistochemical, HE staining, and ELISA for AMA were examined. Results: 1) H.E. staining showed the grade of portal cellular infiltration in the group administered both anti-VLA-4 mAbs and anti-VCAM-1 mAbs was significantly suppressed compared with the control administered normal rat IgG. The grade in the group administered just anti-VLA-4 mAbs was also suppressed compared with the control. 2) The induction of GVHR and the elevation of AMA titer were not affected in these groups by administering mAbs. 3) The expressions of IL-2, IFN'/, IL-4 and IL-10 mRNA were not changed in these groups. 4) Immunohistochemically, CD4, CDS, B220 or Mac-1 positive cells were detected in these groups. Conclusion: Although the administration against adhesion molecules did not suppress the induction of GVHR and the elevation of AMA titer, the portal cellular infiltration is reduced by just the administration of antibodies against VLA-4 in this model. However, cytokine profile of liver infiltrating CD4 ÷ T ceils and the presence of infiltrating ceils such as CD4, CD8, B220 or Mac-1 positive cells were not changed. In conclusion, it is suggested that the administration of antibodies against adhesion molecule could suppress PBC-like lesions without affecting Th 1/Th2 balance. L0272 IMPROVEMENT OF RESPONSE RATE TO IFN BY PATIENTSELECTION THROUGH TESTING GENOTYPES AND NS5A AA22092248 OF HCV RNA BEFORE THERAPY IN PATIENTS WITH CHRONIC HEPATITIS C. Izumi N, Enomoto N*, Watanabe H, Shimizu M, Noguchi O, Hoshino Y, Uchihara M, Miyake S, Sakai T, Sato C#. Division of Gastroenterology and Hepatology, Musashino Red-cross Hospital, *2nd Dept. of Int. Med., #Division of Health Science, Tokyo Medical and Dental University, Tokyo, Japan. We have reported that the response to IFN is closely associated with ISDR (NS5A aa2209-2248, Interferon Sensitivity Determining Region) type in genotype lb chronic hepatitis C. To improve response rate of IFN in patients with chronic hepatitis C, we conducted a prospective study by patient selection before IFN therapy. To the patients with chronic hepatitis C who refered to Musashino Red-Cross Hospital from July, 1996 to October, 1997, genotypes and ISDR types of HCVRNA were tested by the method described by Enomoto et ai. 199 patients were screened, and the distribution of the patients of genotypes la, lb, 2a, 2b and lb+2b were 1, 114, 31, 27 and 1, respectively. In the patients with genotype lb infection, ISDR type was examined, and they were divided into three groups; 55 with the wild type (no amino acid substitution in ISDR), 42 with the intermediate type (1-3 amino acid substitutions)and 15 with the mutant type (above 4 amino acid substitutions). Depending on these results, IFN therapy was recomended mainly to patients with genotype lb-ISDR mutant type, genotype 2a, and 2b infection. IFN therapy was performed to 63 patients after liver biopsy and obtaining infromed consent. Genotypes and ISDR types of the patients who received IFN were as follows; in genotype lb, wild type 6, intermediate type 14, mutant type 7, genotype 2a 20, and genotype 2b 15 patients. 10MU of natural IFN alpha (OIF) was administered everyday for 4 weeks followed by 3 times a week for 20 weeks to all the patients included in this study, and a total dose of IFN was 880MU. Follow-up period for 24 weeks after discontinuation of IFN was completed in 26 patients, and in 13 of them, HCVRNA was no more detectable during the observation period (complete response rate; 50%). Before patient selection system, complete response rate was as high as 29.5% in 431 patients with chronic hepatitis C who received IFN therapy (mean of total dose of IFN; 685MU) in our hospital. In conclusion, it is possible to improve response rate to IFN by patient selection
GASTROENTEROLOGY Vol. 114, No. 4
through testing genotypes and ISDR types of HCV RNA before the therapy in patients with chronic hepatitis C. L0273
EFFECT OF HEPATITIS C TYPE 1B ON PREGNANCY. T. Jabeen, B. Cannon, D. Jenkins, M.J. Whelton. Depts. of Gastroenterology and Gynecology, Cork University Hospital, Ireland. We have studied a unique cohort of Irish women with HCV RNA acquired from contaminated Anti-D immunoglobulin to see if the spontaneous abortion rate increased after infection. METHODS: We studied 36 caucasion primagravida who became infected in 1977. The diagnosis was confirmed by RIBA 3 testing. 28 were HCV RNA positive by PCR when the diagnosis was made in 1994. The group was compared to an age matched Rhesus positive control group who had their first pregnancy in 1977. RESULTS: In the 20 years post-infection the infected group of women had 132 pregnancies (mean parity 3.5). There were 12 spontaneous abortions only one of which occurred in the pregnancy immediately following infection. Birth weights of the 118 children bum at term ranged from 5.3 lbs to 10.5 lbs and did not differ significantly from the control group. Four premature births occurred in the infected group (gestation 32 - 36 weeks). CONCLUSION: In a controlled retrospective study of women with chronic HCV RNA due to infected Anti-D immunoglobulin we have shown no increase in spontaneous abortion. Birth weight of the infants of infected mothers did not differ significantly from those in the control group. L0274
NON-ALCOHOLIC STEATOHEPATITIS (NASH) AND HEREDITARY HEMOCItROMATOSIS (HHC). O Jawaid. P Leclair, K Tortorelli, J Cobb, R Lambrecht, H Bonkovsky. Univ. of Mass Med Ctr, Worcester, MA. NASH is increasingly recognized, and its pathogenesis is believed to involve oxidative stress. Elevated levels of serum ferritin and positive liver iron stains are often observed in patients with NASH, and the pathogenesis of iron related liver injury is also thought to involve oxidative stress. The aim of this study was to determine whether there is an association of NASH and HHC and whether there are differences among NASH patients who do or do not carry the common HFE mutation of HHC (C282Y). Methods: 29 Caucasian patients with NASH (27 biopsy proven) were genotyped by PCR of DNA from peripheral blood by RFLP analysis using SnaB1. Clinical and laboratory data were obtained from medical records and patients interviewed when possible. Histological features of liver biopsies were graded by semiquantitative scales. Data were analyzed by both parameteric and non-parametric methods with similar results. Results: 6 patients (21%) were hh (n=l) or Hh (n=5) for C282Y compared with 6.7% (N Engl J Med 1988; 318:1355-62) of Caucasians (p<0.002). Sex (63-67% male) and age at diagnosis did not differ between the groups. As shown in the table, NASH patients with the mutant (h) gene had significantly higher serum irons and TF saturations, and a tendency toward more stainable hepatic iron. Other features were similar between the two groups (Table). Sex (M/F) (n) Age at diagnosis (y) Transferrin Saturation Serum iron (ug/dL) Ferritin (ng/mL) AST (U/L) ALT (U/L) AP'ase (U/L) Bx iron (0-4+) Bx fat (0-4+) Bx inflammation Bx fibrosis (0-4+)
HH
Hh/hh
p value
15/8 48.0 0.275 85.5 386. 56.7 74 94 0.9 2.4 1.3 0.9
4/2 48.1 0.444 129. 435 54 84 94 1.4 2.6 1 0.8
0.97 0.03 0.02 0.82 0.89 0.65 0.98 0.27 0.68 0.67 0.84
We conclude that there is an increased prevalence of the C282Y mutation of HFE in patients with NASH. We speculate that iron plays a role in pathogenesis of NASH but does not appreciably influence the hepatic histopatho!ogy or lab measures of hepatic injury. • L0275 THE USE OF RECOMBINANT FACTOR VIIA (rFVIIA) IN LAPAROSCOPIC LIVER BIOPSY (LB): A PILOT TRIAL. LJ Jeffers 1, DE Bemstein2, E Erhardsten 3, W Acebo 1, KR Reddy 1, RM Bech 3, ER SchiffI. University of Miami School of Medicine-VA Medical Center, Miami, FL2, Winthrop-University Hospital, Mineola, NY2; Novo Nordisk AS, Copenhagen, DK3. A single intravenous dose of rFVIIa has been shown to correct the prolonged prothrombin time (PT) in non-bleeding, volunteer cirrhotic patients. In this trial, we evaluated the use of a single 5 ug/kg dose of rFVIIa in cirrhotic patients with prolonged prothrombin times undergoing laparoscopic liver
• L0271 SUPPRESSION OF PBC-LIKE HEPATIC LESIONS IN MURINE GVHR M O D E L BY ANTIBODIES AGAINST ADHESION MOLECULES. S. Itoh, Y. Matsuzaki, T. Kimura, T. Ikegami, J. Shoda, N. Tanaka Tsukuba, Ibaraki, Japan. Background: We have previously demonstrated that Thl type cytokine
mRNA firstly increased and Th2 type cytokine mRNA elevated subsequently in liver infiltrating CD4 ÷ T cells of PBC-like hepatic lesion using murine GVHR (Gastroenterology, 110, A1219, 1996). A number of cell adhesion molecules are important for CD4 ÷ T ceils to infiltrate the liver. Moreover, we have also shown that antibodies against adhesion molecules such as LFA-1 and VLA-4 suppressed the formation of PBC-like lesions (Hepatology, 22 (4) pt.2, 1995, Gastroenterology 110 (4) A1235, 1996). However, the precise molecular mechanism of these suppressions by the administration of antibodies against adhesion molecules still remains unknown. Aim: To clarify whether Thl/Th2 balance is related to the suppression of PBC-like hepatic lesions by antibodies against adhesion molecules. Methods: Sex-matched 1 x 107 C57BL/6 (B6) T spleen cells were injected into (B6.C-H-2 bml2 x B6) F 1 mice intravenously. Each 50 lag of anti-mouse t~ chain of VLA-4 mAb (PS2.3), anti-mouse VCAM-1 mAb (MK/2) or normal rat IgG were administered intraperitoneally per mouse during cell transfer. All mice were sacrificed 2 weeks after the first cell transfer. 1 x 10-s liver infiltrating Thy 1.2+CD4÷ lymphocytes were sorted using FACS vantage and the RNA was isolated by AGPC method. A semiquantitative RT-PCR was carried out using primers specific for IL-2, IFN~, IL-4, IL-10. Immunohistochemical, HE staining, and ELISA for AMA were examined. Results: 1) H.E. staining showed the grade of portal cellular infiltration in the group administered both anti-VLA-4 mAbs and anti-VCAM-1 mAbs was significantly suppressed compared with the control administered normal rat IgG. The grade in the group administered just anti-VLA-4 mAbs was also suppressed compared with the control. 2) The induction of GVHR and the elevation of AMA titer were not affected in these groups by administering mAbs. 3) The expressions of IL-2, IFN'/, IL-4 and IL-10 mRNA were not changed in these groups. 4) Immunohistochemically, CD4, CDS, B220 or Mac-1 positive cells were detected in these groups. Conclusion: Although the administration against adhesion molecules did not suppress the induction of GVHR and the elevation of AMA titer, the portal cellular infiltration is reduced by just the administration of antibodies against VLA-4 in this model. However, cytokine profile of liver infiltrating CD4 ÷ T ceils and the presence of infiltrating ceils such as CD4, CD8, B220 or Mac-1 positive cells were not changed. In conclusion, it is suggested that the administration of antibodies against adhesion molecule could suppress PBC-like lesions without affecting Th 1/Th2 balance. L0272 IMPROVEMENT OF RESPONSE RATE TO IFN BY PATIENTSELECTION THROUGH TESTING GENOTYPES AND NS5A AA22092248 OF HCV RNA BEFORE THERAPY IN PATIENTS WITH CHRONIC HEPATITIS C. Izumi N, Enomoto N*, Watanabe H, Shimizu M, Noguchi O, Hoshino Y, Uchihara M, Miyake S, Sakai T, Sato C#. Division of Gastroenterology and Hepatology, Musashino Red-cross Hospital, *2nd Dept. of Int. Med., #Division of Health Science, Tokyo Medical and Dental University, Tokyo, Japan. We have reported that the response to IFN is closely associated with ISDR (NS5A aa2209-2248, Interferon Sensitivity Determining Region) type in genotype lb chronic hepatitis C. To improve response rate of IFN in patients with chronic hepatitis C, we conducted a prospective study by patient selection before IFN therapy. To the patients with chronic hepatitis C who refered to Musashino Red-Cross Hospital from July, 1996 to October, 1997, genotypes and ISDR types of HCVRNA were tested by the method described by Enomoto et ai. 199 patients were screened, and the distribution of the patients of genotypes la, lb, 2a, 2b and lb+2b were 1, 114, 31, 27 and 1, respectively. In the patients with genotype lb infection, ISDR type was examined, and they were divided into three groups; 55 with the wild type (no amino acid substitution in ISDR), 42 with the intermediate type (1-3 amino acid substitutions)and 15 with the mutant type (above 4 amino acid substitutions). Depending on these results, IFN therapy was recomended mainly to patients with genotype lb-ISDR mutant type, genotype 2a, and 2b infection. IFN therapy was performed to 63 patients after liver biopsy and obtaining infromed consent. Genotypes and ISDR types of the patients who received IFN were as follows; in genotype lb, wild type 6, intermediate type 14, mutant type 7, genotype 2a 20, and genotype 2b 15 patients. 10MU of natural IFN alpha (OIF) was administered everyday for 4 weeks followed by 3 times a week for 20 weeks to all the patients included in this study, and a total dose of IFN was 880MU. Follow-up period for 24 weeks after discontinuation of IFN was completed in 26 patients, and in 13 of them, HCVRNA was no more detectable during the observation period (complete response rate; 50%). Before patient selection system, complete response rate was as high as 29.5% in 431 patients with chronic hepatitis C who received IFN therapy (mean of total dose of IFN; 685MU) in our hospital. In conclusion, it is possible to improve response rate to IFN by patient selection
GASTROENTEROLOGY Vol. 114, No. 4
through testing genotypes and ISDR types of HCV RNA before the therapy in patients with chronic hepatitis C. L0273
EFFECT OF HEPATITIS C TYPE 1B ON PREGNANCY. T. Jabeen, B. Cannon, D. Jenkins, M.J. Whelton. Depts. of Gastroenterology and Gynecology, Cork University Hospital, Ireland. We have studied a unique cohort of Irish women with HCV RNA acquired from contaminated Anti-D immunoglobulin to see if the spontaneous abortion rate increased after infection. METHODS: We studied 36 caucasion primagravida who became infected in 1977. The diagnosis was confirmed by RIBA 3 testing. 28 were HCV RNA positive by PCR when the diagnosis was made in 1994. The group was compared to an age matched Rhesus positive control group who had their first pregnancy in 1977. RESULTS: In the 20 years post-infection the infected group of women had 132 pregnancies (mean parity 3.5). There were 12 spontaneous abortions only one of which occurred in the pregnancy immediately following infection. Birth weights of the 118 children bum at term ranged from 5.3 lbs to 10.5 lbs and did not differ significantly from the control group. Four premature births occurred in the infected group (gestation 32 - 36 weeks). CONCLUSION: In a controlled retrospective study of women with chronic HCV RNA due to infected Anti-D immunoglobulin we have shown no increase in spontaneous abortion. Birth weight of the infants of infected mothers did not differ significantly from those in the control group. L0274
NON-ALCOHOLIC STEATOHEPATITIS (NASH) AND HEREDITARY HEMOCItROMATOSIS (HHC). O Jawaid. P Leclair, K Tortorelli, J Cobb, R Lambrecht, H Bonkovsky. Univ. of Mass Med Ctr, Worcester, MA. NASH is increasingly recognized, and its pathogenesis is believed to involve oxidative stress. Elevated levels of serum ferritin and positive liver iron stains are often observed in patients with NASH, and the pathogenesis of iron related liver injury is also thought to involve oxidative stress. The aim of this study was to determine whether there is an association of NASH and HHC and whether there are differences among NASH patients who do or do not carry the common HFE mutation of HHC (C282Y). Methods: 29 Caucasian patients with NASH (27 biopsy proven) were genotyped by PCR of DNA from peripheral blood by RFLP analysis using SnaB1. Clinical and laboratory data were obtained from medical records and patients interviewed when possible. Histological features of liver biopsies were graded by semiquantitative scales. Data were analyzed by both parameteric and non-parametric methods with similar results. Results: 6 patients (21%) were hh (n=l) or Hh (n=5) for C282Y compared with 6.7% (N Engl J Med 1988; 318:1355-62) of Caucasians (p<0.002). Sex (63-67% male) and age at diagnosis did not differ between the groups. As shown in the table, NASH patients with the mutant (h) gene had significantly higher serum irons and TF saturations, and a tendency toward more stainable hepatic iron. Other features were similar between the two groups (Table). Sex (M/F) (n) Age at diagnosis (y) Transferrin Saturation Serum iron (ug/dL) Ferritin (ng/mL) AST (U/L) ALT (U/L) AP'ase (U/L) Bx iron (0-4+) Bx fat (0-4+) Bx inflammation Bx fibrosis (0-4+)
HH
Hh/hh
p value
15/8 48.0 0.275 85.5 386. 56.7 74 94 0.9 2.4 1.3 0.9
4/2 48.1 0.444 129. 435 54 84 94 1.4 2.6 1 0.8
0.97 0.03 0.02 0.82 0.89 0.65 0.98 0.27 0.68 0.67 0.84
We conclude that there is an increased prevalence of the C282Y mutation of HFE in patients with NASH. We speculate that iron plays a role in pathogenesis of NASH but does not appreciably influence the hepatic histopatho!ogy or lab measures of hepatic injury. • L0275 THE USE OF RECOMBINANT FACTOR VIIA (rFVIIA) IN LAPAROSCOPIC LIVER BIOPSY (LB): A PILOT TRIAL. LJ Jeffers 1, DE Bemstein2, E Erhardsten 3, W Acebo 1, KR Reddy 1, RM Bech 3, ER SchiffI. University of Miami School of Medicine-VA Medical Center, Miami, FL2, Winthrop-University Hospital, Mineola, NY2; Novo Nordisk AS, Copenhagen, DK3. A single intravenous dose of rFVIIa has been shown to correct the prolonged prothrombin time (PT) in non-bleeding, volunteer cirrhotic patients. In this trial, we evaluated the use of a single 5 ug/kg dose of rFVIIa in cirrhotic patients with prolonged prothrombin times undergoing laparoscopic liver