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Rare coexpression of tissue HBV and HCV antigens in chronic liver disease: Evidence for viral interference?
HEPATOLOGY Vol. 22, No. 4, Pt. 2, 1995
633
AASLD ABSTRACTS
RESTRICTION OF V BETA GENE USAGE OF LIVER-DERIVED L Y M P H O C Y T E S IN C H R O N I C H E P A T I T I S B A N D C. BN Pham,
634
JF Mosnier. A Sauvanet. S Erlinaer. JHM Cohen* and F Deaos. Service d'Hematologie et Immunologic, Service d'H~patologie, Service d'Anatomie-Pathologique, Service de Chirurgie Digestive, HSpital Beaujon, Clichy, *Laboratoire d'lmmunologie, Htpital Robert Debr~, Reims, France.
WHAT ARE EPIDEMIOLOGICAL, BIOCHEMICAL, VIROLOGICAL AND HISTOLOGIC CHARACTERISTICS OF CHRONIC HEPATITIS BC ? JP. Zarski 1,3, B.Bohn I , A.Bastie2, M.Baud 3, JM.Pawlotsky2, J.Tran Van Nhieu2~ C.Buffet 4, D.Dhumeaux z. Services HtpatoGastroenttrologie-. 1-CHU Grenoble-2-Crtteil-4-Kremlin-Bicetre-3Virologic Grenoble. FRANCE. The purpose of the study was to compare epidemiological, biochemical, virological and histologic characteristics of patients with chronic hepatitis ("CH") B and C by comparison to patients with "CH" C. Hundred eight patients anti-HCV (+), Elisa 3 and Riba 3, anti-HIV (-), anti-HDV (-) apparied on age, sex and mode of contamination were studied and separated in 2 groups according to viral status, [group 1 (N = 54) "CH" BC (HBs Ag (+), HBe Ag = 8, HBe Ab = 46) and group 2 (N = 54)] "CH" C. HBV DNA and HCV RNA were detected in the serum by bDNA and PCR and in the liver for 10 co-infected patients. The semtype was detected by Chirou Riba HCV serotyping SIA technic. Histologic characteristics included Knodeil and Metavir score. HCV + HBV
HCV
m±ISD N=54 N=54 Age (years) 40.6 38.2 NS Sex (M/F) 46 / 8 38 / 16 NS D.A./transfusion 26 / 16 27 / 20 NS ALT (N) 4.4 4.3 NS GGT (N) 2.27 1.4 p = 0.013 bDNA HCV (+) (%) 33 % 79 % p = 0.001 105 Eq/ml 48.5 48.5 NS HCV RNA PCR (%) 53 % 83 % NS Serotype 1 36 % 39 % NS Serotype 3 23 % 11% NS KnodeIl 9.1 6.7 p = 0.001 Piecemeal necrosis 2.14 1.49 p = 0.02 Fibrosis 2.3 1.1 p = 0.001 HBV DNA was detected in the serum of 2 patients by bDNA and 3 by PCR, but for all patients in the liver. These results suggest that in patients with "CH" B-C : 1°) the HCV replication is less frequent but not quantitatively different. 2 °) the HBV replication is very rare in the serum but ever present in the liver. 3 °) the serotype is not different. 4 °) the histologic lesions are more severe.
I N T E R F E R E N C E IN D O U B L E I N F E C T I O N OF H E P A T I T I S B AND C V I R U S . H Tsuii. H Shimomura. MWato. J Kondo, T Hasui, K Fujio, Y lshii, S Fujioka. T Doi. G Yamada. and T Tsuji First Department of Internal Medicine, Okayama University Medical School, Okayama, Japan. VIRAL
The possibility of interference in viral replication between hepatitis B virus (HBV) and hepatitis C virus grlCV) has been reported, but most of thereports were merely based on the qualitative assays for viral markers. Here we report the quantitative analysis of viral markers for HBV and HCV to clarify mutual viral influence. M E T H O D S : Twenty-seven chronic hepatitis patients positive for both HBsAg and anti-HCV were studied (M/F 20/7; age 55--+13). Six patients positive for HBeAg or high titer of DNA polymerase were grouped as Group BH and the others as Group BL. Twenty-one patients positive for HBsAg and anti-HBe and negative for anti-HCV were chosen to Group BC as control, whose age and gender matched to Group BL. Group CC, 116 patients with hepatitis C alone, were used as another control. HBsAg were quantified by ELISA, HBV-associated DNA poiymerase by RA, pre-S2 antigen by polymerized human albumin binding assay (ELISA), HBV-DNA by competitive PCR, and HCV-RNA by competitive RT-PCR ( log (copy / 0.5 ml) unit ). R E S U L T : HCV-RNA was detected in 2 (33%) of Group BH, and 17 (81%) of Group BL. "liter of HCV-RNA of Group BL (6.9-+0.7) tended to be lower than that of Group CC (7.4+-0.8). HBV-DNA was detected in 9 (42.9%) of Group BL and 11 (52.4%) of Group BC. Amount of pre-S2 were not different between Group BL and BC, but quantity of HBsAg tended to be decreased in Group BL than Group B C (7.77 -+ 6.62 vs. 10.44 -+ 6.84, respectively). CONCLUSION: It is suggested that replication of HCV was suppressed by HBV not only at HBeAg-positive phase, but at anti-HBepositive phase, in cases of double infection of HBV and HCV. On the other hand, HCV seemed to suppress the replication of HBV as seen in Group BL.
T lymphocytes have been reported to be the predominant inflammatory cells in the liver of patients with chronic viral hepatitis. Their presence may reflect either non-specific inflammation related to the persistence of the virus in the liver cells or a virus-specific immune response. A restricted usage of T cell receptor (TCR) V gene segments is currently interpreted as indicating the specificity of a T ceil response. To assess the repertoire of intra-hepatic T cells, we investigated the TCR V beta gene usage of T cells in 6 patients with chronic hepatitis B and 9 with chronic hepatitis C. Liver-derived lymphocytes and peripheral blood lymphocytes were analyzed by flow cytometry using a panel of 6 V~ gene-specific anti-TCR monoclonal antibodies. In patients with chronic hepatitis B, 2 out of 6 patients were found to have an accumulation of V~ 6.7 T cells in the liver. In patients with chronic hepatitis C, 2 out of 9 patients were found to.have an accumulation of VI3 5.1 T cells orVl3 8 and VI3 5.2 and 5.3 T cells in the liver respectively. In the liver of the other patients, we found no accumulation of T cells using the analyzed VI3. Despite a limited screening of V beta subfamilies, this study indicates that, in patients with chronic hepatitis B as well as in patients with chronic hepatitis C, T cells using a certain V beta gene may accumulate in the liver. This suggests that intra-hepatic T cells are oligoclonal and possibly virus-specific. Our results argue against the role of a superantigen in perpetuating liver disease. In addition, this study supports a role for T lymphocytes in the pathogenesis of chronic hepatitis C.
635
265A
636
RARE COEXPRESStON OF TISSUE HBV AND HCV ANTIGENS IN CHRONIC LIVER DISEASE: EVIDENCE FOR VIRAL INTERFERENCE? G Ballardini~ P Croft, F Gioatra v R Franceeconi~ #A Manzin~ *R Miniero, S Gheftiv A Grassi~ D Zauli~ FB Bianchi. Semeiotica Medica II, *Laboratorio Centr. Pol. S.Orsola, University of Bologna; #1stituto di Virologia, University of Ancona, Italy Both increased HBsAg clearance after HCV infection and absence of HCV viremia in HBeAg positive patients have been reported. To further evaluate the presence of viral interference we retrospectively studied liver HCV antigens expression in 11 HI3sAg+, anti-HCV+, RIBA3+ (of which 5 HBeAg+, 6 anti-HBe+ and 3 anti-Delta+) and 100 HBsAg- (of which 30 antiHBs+ and 70 anti-HBs-) anti-HCV+ patients with chronic liver disease. HCV RNA was positive in SO out of the 84 HBsAg- and in none of the 3 HBsAg+ cases tested. Tissue HBsAg and core antigens were present in 9111 and 4/11 HBsAg+ cases by immunohistochemiatry, respectively. Hepatocellular HCV antigens were searched for on frozen liver sections using FITC-conjugated IgG from high titer anti-HCV positive sera, followed by mouse anti-FITC, rabbit anti-mouse-PX and swine anti-rabbit-PX. Positivity was visually scored on the basis of % positive hepatocytes from 0 to 3. The reliability of the score system was confirmed by significant correlation (r=0.734) with the morphometric evaluation performed in 22 positive cases. patients n. tissue HCV+ mean score SD HCV+ HBsAg+ 11 1 (9%) HCV+ anti-HBs+ 30 20 (67%) 1.20 1.06 HCV+ anti-NSs- 70 56 (83%) 1.80 1.23 Significant differences were found between HBsAg+ and the other two groups (I)<6.002 and p<0.0000, respectively). The difference between anti-HBs+ and anti-HRs- was not significant in terms of prevalence of positivity. However in the latter group the number of positive hepatocytes (mean score) was higher (p
633
AASLD ABSTRACTS
RESTRICTION OF V BETA GENE USAGE OF LIVER-DERIVED L Y M P H O C Y T E S IN C H R O N I C H E P A T I T I S B A N D C. BN Pham,
634
JF Mosnier. A Sauvanet. S Erlinaer. JHM Cohen* and F Deaos. Service d'Hematologie et Immunologic, Service d'H~patologie, Service d'Anatomie-Pathologique, Service de Chirurgie Digestive, HSpital Beaujon, Clichy, *Laboratoire d'lmmunologie, Htpital Robert Debr~, Reims, France.
WHAT ARE EPIDEMIOLOGICAL, BIOCHEMICAL, VIROLOGICAL AND HISTOLOGIC CHARACTERISTICS OF CHRONIC HEPATITIS BC ? JP. Zarski 1,3, B.Bohn I , A.Bastie2, M.Baud 3, JM.Pawlotsky2, J.Tran Van Nhieu2~ C.Buffet 4, D.Dhumeaux z. Services HtpatoGastroenttrologie-. 1-CHU Grenoble-2-Crtteil-4-Kremlin-Bicetre-3Virologic Grenoble. FRANCE. The purpose of the study was to compare epidemiological, biochemical, virological and histologic characteristics of patients with chronic hepatitis ("CH") B and C by comparison to patients with "CH" C. Hundred eight patients anti-HCV (+), Elisa 3 and Riba 3, anti-HIV (-), anti-HDV (-) apparied on age, sex and mode of contamination were studied and separated in 2 groups according to viral status, [group 1 (N = 54) "CH" BC (HBs Ag (+), HBe Ag = 8, HBe Ab = 46) and group 2 (N = 54)] "CH" C. HBV DNA and HCV RNA were detected in the serum by bDNA and PCR and in the liver for 10 co-infected patients. The semtype was detected by Chirou Riba HCV serotyping SIA technic. Histologic characteristics included Knodeil and Metavir score. HCV + HBV
HCV
m±ISD N=54 N=54 Age (years) 40.6 38.2 NS Sex (M/F) 46 / 8 38 / 16 NS D.A./transfusion 26 / 16 27 / 20 NS ALT (N) 4.4 4.3 NS GGT (N) 2.27 1.4 p = 0.013 bDNA HCV (+) (%) 33 % 79 % p = 0.001 105 Eq/ml 48.5 48.5 NS HCV RNA PCR (%) 53 % 83 % NS Serotype 1 36 % 39 % NS Serotype 3 23 % 11% NS KnodeIl 9.1 6.7 p = 0.001 Piecemeal necrosis 2.14 1.49 p = 0.02 Fibrosis 2.3 1.1 p = 0.001 HBV DNA was detected in the serum of 2 patients by bDNA and 3 by PCR, but for all patients in the liver. These results suggest that in patients with "CH" B-C : 1°) the HCV replication is less frequent but not quantitatively different. 2 °) the HBV replication is very rare in the serum but ever present in the liver. 3 °) the serotype is not different. 4 °) the histologic lesions are more severe.
I N T E R F E R E N C E IN D O U B L E I N F E C T I O N OF H E P A T I T I S B AND C V I R U S . H Tsuii. H Shimomura. MWato. J Kondo, T Hasui, K Fujio, Y lshii, S Fujioka. T Doi. G Yamada. and T Tsuji First Department of Internal Medicine, Okayama University Medical School, Okayama, Japan. VIRAL
The possibility of interference in viral replication between hepatitis B virus (HBV) and hepatitis C virus grlCV) has been reported, but most of thereports were merely based on the qualitative assays for viral markers. Here we report the quantitative analysis of viral markers for HBV and HCV to clarify mutual viral influence. M E T H O D S : Twenty-seven chronic hepatitis patients positive for both HBsAg and anti-HCV were studied (M/F 20/7; age 55--+13). Six patients positive for HBeAg or high titer of DNA polymerase were grouped as Group BH and the others as Group BL. Twenty-one patients positive for HBsAg and anti-HBe and negative for anti-HCV were chosen to Group BC as control, whose age and gender matched to Group BL. Group CC, 116 patients with hepatitis C alone, were used as another control. HBsAg were quantified by ELISA, HBV-associated DNA poiymerase by RA, pre-S2 antigen by polymerized human albumin binding assay (ELISA), HBV-DNA by competitive PCR, and HCV-RNA by competitive RT-PCR ( log (copy / 0.5 ml) unit ). R E S U L T : HCV-RNA was detected in 2 (33%) of Group BH, and 17 (81%) of Group BL. "liter of HCV-RNA of Group BL (6.9-+0.7) tended to be lower than that of Group CC (7.4+-0.8). HBV-DNA was detected in 9 (42.9%) of Group BL and 11 (52.4%) of Group BC. Amount of pre-S2 were not different between Group BL and BC, but quantity of HBsAg tended to be decreased in Group BL than Group B C (7.77 -+ 6.62 vs. 10.44 -+ 6.84, respectively). CONCLUSION: It is suggested that replication of HCV was suppressed by HBV not only at HBeAg-positive phase, but at anti-HBepositive phase, in cases of double infection of HBV and HCV. On the other hand, HCV seemed to suppress the replication of HBV as seen in Group BL.
T lymphocytes have been reported to be the predominant inflammatory cells in the liver of patients with chronic viral hepatitis. Their presence may reflect either non-specific inflammation related to the persistence of the virus in the liver cells or a virus-specific immune response. A restricted usage of T cell receptor (TCR) V gene segments is currently interpreted as indicating the specificity of a T ceil response. To assess the repertoire of intra-hepatic T cells, we investigated the TCR V beta gene usage of T cells in 6 patients with chronic hepatitis B and 9 with chronic hepatitis C. Liver-derived lymphocytes and peripheral blood lymphocytes were analyzed by flow cytometry using a panel of 6 V~ gene-specific anti-TCR monoclonal antibodies. In patients with chronic hepatitis B, 2 out of 6 patients were found to have an accumulation of V~ 6.7 T cells in the liver. In patients with chronic hepatitis C, 2 out of 9 patients were found to.have an accumulation of VI3 5.1 T cells orVl3 8 and VI3 5.2 and 5.3 T cells in the liver respectively. In the liver of the other patients, we found no accumulation of T cells using the analyzed VI3. Despite a limited screening of V beta subfamilies, this study indicates that, in patients with chronic hepatitis B as well as in patients with chronic hepatitis C, T cells using a certain V beta gene may accumulate in the liver. This suggests that intra-hepatic T cells are oligoclonal and possibly virus-specific. Our results argue against the role of a superantigen in perpetuating liver disease. In addition, this study supports a role for T lymphocytes in the pathogenesis of chronic hepatitis C.
635
265A
636
RARE COEXPRESStON OF TISSUE HBV AND HCV ANTIGENS IN CHRONIC LIVER DISEASE: EVIDENCE FOR VIRAL INTERFERENCE? G Ballardini~ P Croft, F Gioatra v R Franceeconi~ #A Manzin~ *R Miniero, S Gheftiv A Grassi~ D Zauli~ FB Bianchi. Semeiotica Medica II, *Laboratorio Centr. Pol. S.Orsola, University of Bologna; #1stituto di Virologia, University of Ancona, Italy Both increased HBsAg clearance after HCV infection and absence of HCV viremia in HBeAg positive patients have been reported. To further evaluate the presence of viral interference we retrospectively studied liver HCV antigens expression in 11 HI3sAg+, anti-HCV+, RIBA3+ (of which 5 HBeAg+, 6 anti-HBe+ and 3 anti-Delta+) and 100 HBsAg- (of which 30 antiHBs+ and 70 anti-HBs-) anti-HCV+ patients with chronic liver disease. HCV RNA was positive in SO out of the 84 HBsAg- and in none of the 3 HBsAg+ cases tested. Tissue HBsAg and core antigens were present in 9111 and 4/11 HBsAg+ cases by immunohistochemiatry, respectively. Hepatocellular HCV antigens were searched for on frozen liver sections using FITC-conjugated IgG from high titer anti-HCV positive sera, followed by mouse anti-FITC, rabbit anti-mouse-PX and swine anti-rabbit-PX. Positivity was visually scored on the basis of % positive hepatocytes from 0 to 3. The reliability of the score system was confirmed by significant correlation (r=0.734) with the morphometric evaluation performed in 22 positive cases. patients n. tissue HCV+ mean score SD HCV+ HBsAg+ 11 1 (9%) HCV+ anti-HBs+ 30 20 (67%) 1.20 1.06 HCV+ anti-NSs- 70 56 (83%) 1.80 1.23 Significant differences were found between HBsAg+ and the other two groups (I)<6.002 and p<0.0000, respectively). The difference between anti-HBs+ and anti-HRs- was not significant in terms of prevalence of positivity. However in the latter group the number of positive hepatocytes (mean score) was higher (p