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Distinct regulation of MHC class I in human cytomegalovirus infected epithelial cells of hepatobiliary and intestinal origin
April 2000
based on the data for serial three-years series over 12 years. The median of the age of male patients of HCC with HCV were 59, 60, 64, 66 years, and the that of female patients were 62.5, 63, 69, 68.5 years. The number of male patients over 70 years of age were 3 (10%), 11 (13%), 16 (23%), 24 (27%), and also that of female patients over 70 years of age were 3 (17%), 8 (26%), 16 (48%), 17 (47%). These findings identify senile patients of HCC with HCV are increasing. There was no difference between patients over 70 years of age and patients under 70 years of age about the tumor staging and the factors of background. Discussion/Conclusion: In all Americans, the highest prevalences ofHCV infection were noted in persons aged 30 to 50 years old, while these were noted in persons aged 50 to 70 years old in Japan. In this study, a major peak of HCC associated with HCV was obviously advanced without distinct of sex, and the number of HCC associated with HCV of patients over 70 years of age was roughly similar in both genders. There is difference of a major peak of HCV infection between all Americans and Japanese, and seroprevalence increases with advancing age. Thus, the Japanese experience might precede the American experience by 2-3 decades. While the rate of death due to HCC was generally far often among men, these results indicate that senile women associated with HCV might have a potential risk for HCC as well as men. Patients of HCV-related chronic liver disease should be closely followed up by various examinations regardless of sex.
263 DISTINCT REGULATION OF MHC CLASS I IN HUMAN CYTO· MEGALOVIRUS INFECTED EPITHELIAL CELLS OF HEPATOBILIARY AND INTESTINAL ORIGIN. Christine Benz, Uwe Reusch, Walter Muranyi, Hartmut Hengel, Univ of Medicine, Koeln, Germany; Max von Pettenkofer-Institut, Muenchen, Germany. Liver and intestinal epithelial cells are major targets of infection by human cytomegalovirus (HCMV) causing severe disease in affected organs of immunocompromised patients. HCMV downregulates major histocompatibility complex class I (MHC I) molecules in fibroblasts to prevent antigen presentation to CD8+ cytotoxic T lymphocytes. However, MHC class I expression in HCMV-infected hepatic tissue of patients was reported to be increased. As it is unclear at present whether HCMV affects MHC I in cells other than fibroblasts, we established new cell culture models for HCMV infection of differentiated hepatobiliary and colonic cell lines. HCMV gene expression was achieved in 60 to 95 % of cells but replication differed from prototypic infection of fibroblasts since HCMV progeny yielded significantly lower titers. HCMV infection resulted in a drastic and selective downregulation of MHC I which was uniformly observed in hepatic, biliary and colonic epithelial cells. All known MHC I downregulating HCMV factors, i.e. gpUS2, gpUS3, gpUS6 and gpUS1I were synthesized with similar kinetics and in comparable amounts as in fibroblasts. However, interferon-y (IFN-')') rescued MHC I expression in preinfected epithelial cells but not in fibroblasts. These results demonstrate that HCMV infected tissues show important immunological differences.
264 VIRAL HEPATITIS IS AN INFREQUENT CAUSE OF ELEVATED ALANINE AMINOTRANSFERSE LEVELS IN HEMODIALYSIS PATIENTS. Sammy Saab, Paul Martin, David Ly, Deston Langfield, Maria Brezina, Gary Gitnick, Hal F. Vee, UCLA Sch of Medicine, Los Angeles, CA. Introduction Patients are often screened for viral hepatitis using serum biochemical markers such as alanine aminotransferase (ALT). The sensitivity of acute ALT elevation for de novo viral infection is uncertain. We hypothesized that de novo viral hepatitis is an infrequent cause of ALT elevation in hemodialysis (HD) patients. Our aim was to determine how often acute ALT elevations in HD patients results from de novo hepatitis A, B, and C infection. Methods This prospective study was performed on 2624 HD patients screened by the UCLA Hepatitis Screening Program. Baseline ALT was obtained during 1998 on all patients. ALT was measured monthly and anti-HCV, HBsAg, and anti-HBsAb were checked yearly. Assays for anti-HCV (ELISA-III), HBsAg (radioimmunoassay), anti-HBcAb IgM (ELISA), and anti-HAV IgM (ELISA) were performed if ALT was elevated (>44IU/L). Positive results from an ELISA III antiHCV were confirmed with RIBA III. Results of all HBsAg were confirmed with an enzyme immunoassay. Results. Between January 1, 1998 and November 30, 1999,490 of 2624 patients tested by our program had at least one acute elevated ALT. No patient was found to have de novo hepatitis A or B infection. In addition, no patients underwent HCV seroconversion at the time of acute ALT elevation. Repeat HCV testing performed at least three months after the acute ALT elevation identified 4 patients who developed anti-HCV. No patient subsequently developed HBV infection when followed for at least three months after the ALT elevation. Conclusion Our results indicate that viral hepatitis is a relatively uncommon cause of ALT elevation among HD patients, and consequently bring into question the cost-effectiveness of ALT surveillance in HD patients. HCV testing should be performed at least three months after an acutely elevated ALT because acute ALT elevation is not associated immediately with HCV seroconversion. Future studies should address alternative causes of ALT elevation other than viral hepatitis.
AASLDA957
265 IMMUNOLOGIC ALTERATIONS IN PREGNANT WOMEN WITH HEPATITIS E INFECTION. Rekha PaJ, Subhash R. Naik, Rakesh Aggarwal, Siddharth Das, Vineeta Das, Sita Naik, Sanjay Gandhi Postgraduate Institute of Med Sci, Lucknow, India; KIng George's Med ColI, Lucknow, India. Hepatitis E virus (HEV) infection is more often associated with fulminant liver failure and death in pregnant than in non-pregnant women. The exact cause for this is unclear. We therefore studied non-specific cytotoxic mechanisms of liver cell injury in 10 pregnant women with hepatitis E (P-HEV), 8 pregnant controls (P-C) and 8 non-pregnant healthy controls (NP-C). HEV infection was diagnosed by the presence of IgM anti-HEY antibodies in serum. Median (range) % lysis for natural killer cell (NK) function against K562 targets in the three groups was as follows: P-HEV 28.4 (5.1-74.4), P-C 27.3 (9.1-70.6), and NP-C 52.1 (27.1-69). Percent lysis for antibody-dependent cell cytotoxicity (ADCC) against chicken RBC target was: P-HEV 41.3 (15.1-62.8), P-C 44.2 (0-75.9) and NP-C 5.6 (0-39.2). NK cell activity was significantly lower in P-HEV group (p=0.025; Mann-Whitney U test) and P-C (p=0.008) than NP-C; on the other hand, ADCC was significantly increased in P-HEV (p=0.OO4) and P-C (p=O.OOI) than in NP-C. Thus, NK cell activity was reduced and ADCC activity was enhanced among pregnant women, irrespective of HEV infection. In a set of separate experiments, lymphocyte transformation tests and antigen stimulated IL-4 levels by enzyme immunoassay were done in 13 subjects with P-HEV, 23 with P-C and 8 non-pregnant women with hepatitis E (NP-HEV). A significantly lower lymphoproliferative response to phytohemagglutinin (PHA) was seen in P-HEV group [9.4 (4-460)] than in NP-HEV [49.8 (7.5-313); p=.045] or in P-C [66.1 (7.4367); p= .007]. In response to a mixture of peptides derived from all three open reading frames of HEV, P-HEV had lymphoproliferation [1.45 (04.6)] which was similar to that in NP-HEV [0.88 (0.1-35.5); p=0.12], but higher than that in P-C [0 (0.0-3.7); p=0.042]. Antigen-stimulated IL-4 response was significantly higher in P-HEV [26 (10-310)] than in P-C [0 (0-30)]; p
266 DOPPLER MEASUREMENTS IN PATIENTS WITH CHRONIC VI· RAL HEPATITIS - USEFUL TOOL FOR TREATMENT DECISION? Thomas Bernatik, Deike Strobel, Eckhard G. Hahn, Dirk Becker, Dept of Medicine I, Univ Erlangen-Nurernberg, Erlangen, Germany. Background/Aims: Patients with liver cirrhosis show reduced blood flow in the portal vein, which can be estimated by Doppler measurement of the portal vein blood flow velocity. In patients with chonic viral hepatitis (CVH) it would be of much greater clinical impact to assess the stage of liver fibrosis and grade of inflammatory changes by Doppler imaging. However the potential value of Doppler measurement of all liver vessels in therapeutic decision in patients with CVH has not yet been established. The study focused on the potential correlation between Doppler measurements and histologically proven liver fibrosis and histologically proven inflammation. Materials and Methods: 43 consecutive patients with CVH (79% chronic Hepatitis C) who underwent liver biopsy for grading of inflammatory and fibrotic changes were enrolled in the study. At the time of liver biopsy two independent investigators measured maximum blood flow velocity, mean velocity, resistance index (RI), diameter of the vessel and the blood flow volume in portal vein, hepatic artery and liver veins. All measurements were done by each investigator in triplicate. The mean values were correlated with the degree of liver fibrosis and inflammation graded using the Ludwig-score (Ludwig J: The nomenclature of chronic active hepatitis. Gastroenterology 1993;105:274). Results: 67% of all patients had none or only mild liver fibrosis (LS stage I-II). Between all stages of liver fibrosis and inflammation grades there was a large overlap of all measured doppler parameters. No significant correlation could be found even comparing patients with mild liver fibrosis (LS stage I-II) to patients with severe liver fibrosis (LS stage III-IV). Comparing patients with minimal histologic changes (LS sum 1-2) to patients with futher progressed liver disease (LS sum >2) there was a significant difference in RI in the hepatic artery (p=0,017). Although 92% (12/13) of patients with RI above 0,66 presented further progressed liver disease (LS sum >2), the sensitivity of RI (cutoff 0,66) for indication of severe histological changes (LS sum >2) is only 39% (specifity being 91%). Conclusion: In patients with CVH Doppler measurements neither of the portal vein nor the hepatic artery nor the liver veins are a valid surrogate marker of liver fibrosis or inflammation. Nevertheless increased RI does indicate further progressed histological changes and therefore might indicate need of therapy.
based on the data for serial three-years series over 12 years. The median of the age of male patients of HCC with HCV were 59, 60, 64, 66 years, and the that of female patients were 62.5, 63, 69, 68.5 years. The number of male patients over 70 years of age were 3 (10%), 11 (13%), 16 (23%), 24 (27%), and also that of female patients over 70 years of age were 3 (17%), 8 (26%), 16 (48%), 17 (47%). These findings identify senile patients of HCC with HCV are increasing. There was no difference between patients over 70 years of age and patients under 70 years of age about the tumor staging and the factors of background. Discussion/Conclusion: In all Americans, the highest prevalences ofHCV infection were noted in persons aged 30 to 50 years old, while these were noted in persons aged 50 to 70 years old in Japan. In this study, a major peak of HCC associated with HCV was obviously advanced without distinct of sex, and the number of HCC associated with HCV of patients over 70 years of age was roughly similar in both genders. There is difference of a major peak of HCV infection between all Americans and Japanese, and seroprevalence increases with advancing age. Thus, the Japanese experience might precede the American experience by 2-3 decades. While the rate of death due to HCC was generally far often among men, these results indicate that senile women associated with HCV might have a potential risk for HCC as well as men. Patients of HCV-related chronic liver disease should be closely followed up by various examinations regardless of sex.
263 DISTINCT REGULATION OF MHC CLASS I IN HUMAN CYTO· MEGALOVIRUS INFECTED EPITHELIAL CELLS OF HEPATOBILIARY AND INTESTINAL ORIGIN. Christine Benz, Uwe Reusch, Walter Muranyi, Hartmut Hengel, Univ of Medicine, Koeln, Germany; Max von Pettenkofer-Institut, Muenchen, Germany. Liver and intestinal epithelial cells are major targets of infection by human cytomegalovirus (HCMV) causing severe disease in affected organs of immunocompromised patients. HCMV downregulates major histocompatibility complex class I (MHC I) molecules in fibroblasts to prevent antigen presentation to CD8+ cytotoxic T lymphocytes. However, MHC class I expression in HCMV-infected hepatic tissue of patients was reported to be increased. As it is unclear at present whether HCMV affects MHC I in cells other than fibroblasts, we established new cell culture models for HCMV infection of differentiated hepatobiliary and colonic cell lines. HCMV gene expression was achieved in 60 to 95 % of cells but replication differed from prototypic infection of fibroblasts since HCMV progeny yielded significantly lower titers. HCMV infection resulted in a drastic and selective downregulation of MHC I which was uniformly observed in hepatic, biliary and colonic epithelial cells. All known MHC I downregulating HCMV factors, i.e. gpUS2, gpUS3, gpUS6 and gpUS1I were synthesized with similar kinetics and in comparable amounts as in fibroblasts. However, interferon-y (IFN-')') rescued MHC I expression in preinfected epithelial cells but not in fibroblasts. These results demonstrate that HCMV infected tissues show important immunological differences.
264 VIRAL HEPATITIS IS AN INFREQUENT CAUSE OF ELEVATED ALANINE AMINOTRANSFERSE LEVELS IN HEMODIALYSIS PATIENTS. Sammy Saab, Paul Martin, David Ly, Deston Langfield, Maria Brezina, Gary Gitnick, Hal F. Vee, UCLA Sch of Medicine, Los Angeles, CA. Introduction Patients are often screened for viral hepatitis using serum biochemical markers such as alanine aminotransferase (ALT). The sensitivity of acute ALT elevation for de novo viral infection is uncertain. We hypothesized that de novo viral hepatitis is an infrequent cause of ALT elevation in hemodialysis (HD) patients. Our aim was to determine how often acute ALT elevations in HD patients results from de novo hepatitis A, B, and C infection. Methods This prospective study was performed on 2624 HD patients screened by the UCLA Hepatitis Screening Program. Baseline ALT was obtained during 1998 on all patients. ALT was measured monthly and anti-HCV, HBsAg, and anti-HBsAb were checked yearly. Assays for anti-HCV (ELISA-III), HBsAg (radioimmunoassay), anti-HBcAb IgM (ELISA), and anti-HAV IgM (ELISA) were performed if ALT was elevated (>44IU/L). Positive results from an ELISA III antiHCV were confirmed with RIBA III. Results of all HBsAg were confirmed with an enzyme immunoassay. Results. Between January 1, 1998 and November 30, 1999,490 of 2624 patients tested by our program had at least one acute elevated ALT. No patient was found to have de novo hepatitis A or B infection. In addition, no patients underwent HCV seroconversion at the time of acute ALT elevation. Repeat HCV testing performed at least three months after the acute ALT elevation identified 4 patients who developed anti-HCV. No patient subsequently developed HBV infection when followed for at least three months after the ALT elevation. Conclusion Our results indicate that viral hepatitis is a relatively uncommon cause of ALT elevation among HD patients, and consequently bring into question the cost-effectiveness of ALT surveillance in HD patients. HCV testing should be performed at least three months after an acutely elevated ALT because acute ALT elevation is not associated immediately with HCV seroconversion. Future studies should address alternative causes of ALT elevation other than viral hepatitis.
AASLDA957
265 IMMUNOLOGIC ALTERATIONS IN PREGNANT WOMEN WITH HEPATITIS E INFECTION. Rekha PaJ, Subhash R. Naik, Rakesh Aggarwal, Siddharth Das, Vineeta Das, Sita Naik, Sanjay Gandhi Postgraduate Institute of Med Sci, Lucknow, India; KIng George's Med ColI, Lucknow, India. Hepatitis E virus (HEV) infection is more often associated with fulminant liver failure and death in pregnant than in non-pregnant women. The exact cause for this is unclear. We therefore studied non-specific cytotoxic mechanisms of liver cell injury in 10 pregnant women with hepatitis E (P-HEV), 8 pregnant controls (P-C) and 8 non-pregnant healthy controls (NP-C). HEV infection was diagnosed by the presence of IgM anti-HEY antibodies in serum. Median (range) % lysis for natural killer cell (NK) function against K562 targets in the three groups was as follows: P-HEV 28.4 (5.1-74.4), P-C 27.3 (9.1-70.6), and NP-C 52.1 (27.1-69). Percent lysis for antibody-dependent cell cytotoxicity (ADCC) against chicken RBC target was: P-HEV 41.3 (15.1-62.8), P-C 44.2 (0-75.9) and NP-C 5.6 (0-39.2). NK cell activity was significantly lower in P-HEV group (p=0.025; Mann-Whitney U test) and P-C (p=0.008) than NP-C; on the other hand, ADCC was significantly increased in P-HEV (p=0.OO4) and P-C (p=O.OOI) than in NP-C. Thus, NK cell activity was reduced and ADCC activity was enhanced among pregnant women, irrespective of HEV infection. In a set of separate experiments, lymphocyte transformation tests and antigen stimulated IL-4 levels by enzyme immunoassay were done in 13 subjects with P-HEV, 23 with P-C and 8 non-pregnant women with hepatitis E (NP-HEV). A significantly lower lymphoproliferative response to phytohemagglutinin (PHA) was seen in P-HEV group [9.4 (4-460)] than in NP-HEV [49.8 (7.5-313); p=.045] or in P-C [66.1 (7.4367); p= .007]. In response to a mixture of peptides derived from all three open reading frames of HEV, P-HEV had lymphoproliferation [1.45 (04.6)] which was similar to that in NP-HEV [0.88 (0.1-35.5); p=0.12], but higher than that in P-C [0 (0.0-3.7); p=0.042]. Antigen-stimulated IL-4 response was significantly higher in P-HEV [26 (10-310)] than in P-C [0 (0-30)]; p
266 DOPPLER MEASUREMENTS IN PATIENTS WITH CHRONIC VI· RAL HEPATITIS - USEFUL TOOL FOR TREATMENT DECISION? Thomas Bernatik, Deike Strobel, Eckhard G. Hahn, Dirk Becker, Dept of Medicine I, Univ Erlangen-Nurernberg, Erlangen, Germany. Background/Aims: Patients with liver cirrhosis show reduced blood flow in the portal vein, which can be estimated by Doppler measurement of the portal vein blood flow velocity. In patients with chonic viral hepatitis (CVH) it would be of much greater clinical impact to assess the stage of liver fibrosis and grade of inflammatory changes by Doppler imaging. However the potential value of Doppler measurement of all liver vessels in therapeutic decision in patients with CVH has not yet been established. The study focused on the potential correlation between Doppler measurements and histologically proven liver fibrosis and histologically proven inflammation. Materials and Methods: 43 consecutive patients with CVH (79% chronic Hepatitis C) who underwent liver biopsy for grading of inflammatory and fibrotic changes were enrolled in the study. At the time of liver biopsy two independent investigators measured maximum blood flow velocity, mean velocity, resistance index (RI), diameter of the vessel and the blood flow volume in portal vein, hepatic artery and liver veins. All measurements were done by each investigator in triplicate. The mean values were correlated with the degree of liver fibrosis and inflammation graded using the Ludwig-score (Ludwig J: The nomenclature of chronic active hepatitis. Gastroenterology 1993;105:274). Results: 67% of all patients had none or only mild liver fibrosis (LS stage I-II). Between all stages of liver fibrosis and inflammation grades there was a large overlap of all measured doppler parameters. No significant correlation could be found even comparing patients with mild liver fibrosis (LS stage I-II) to patients with severe liver fibrosis (LS stage III-IV). Comparing patients with minimal histologic changes (LS sum 1-2) to patients with futher progressed liver disease (LS sum >2) there was a significant difference in RI in the hepatic artery (p=0,017). Although 92% (12/13) of patients with RI above 0,66 presented further progressed liver disease (LS sum >2), the sensitivity of RI (cutoff 0,66) for indication of severe histological changes (LS sum >2) is only 39% (specifity being 91%). Conclusion: In patients with CVH Doppler measurements neither of the portal vein nor the hepatic artery nor the liver veins are a valid surrogate marker of liver fibrosis or inflammation. Nevertheless increased RI does indicate further progressed histological changes and therefore might indicate need of therapy.