Vol. 27, No. 5 Printed in Japan
Gastroenterologia Japonica Copyright 9 1992 by The Japanese Society of Gastroenterology
Hepatitis type C virus infection in patients with type B chronic liver disease Toshihiko DOI, Gotaro YAMADA, Hisashi ENDO, Hiroshi NISHIMOTO, Michiko TAKAHASHI, Rieko MIYAMOTO, Shigeatsu FUJIKI, Hiroyuki SHIMOMURA, Motowo MIZUNO, and Takao TSUJI First Department of Internal Medicine, Okayama University Medical School Okayama, Japan Summary: Anti-cl00-3 (Ortho) was determined in the sera of 152 patients with HBs antigen-positive chronic liver diseases to assess coinfection of hepatitis B virus (HBV) and hepatitis C virus (HCV). Eleven patients (7.2%) were positive for anti-cl00-3. Anti-CP-9 (Okamoto) and HCV-RNA (RT-PCR) were also examined in these 11 patients. Anti-CP-9 was detected in 7 patients and HCV-RNA was detected in all 11 patients. Four of the 11 anti-cl00-3-positive patients were positive for HBe antigen (HBeAg) and others were negative. In 8 of the 11 patients, HCV was suspected to be superinfected by blood transfusion. In HBeAgpositive patients, serum glutamic pyruvic transaminase (SGPT) was elevated in relation to active replication of HBV shown by DNA-polymerase activity. The histological findings showed chronic active hepatitis, with or without cirrhosis. On the other hand, in HBeAg-negative patients, SGPT fluctuated without evidence of active replication of HBV. Active inflammation in the liver was observed in 3 of 5 HBeAg-negative patients by liver biopsy. These findings suggest that HBV might play an important role in chronic active inflammation in HBeAg-positive patients coinfected with HCV, and that HCV might be responsible for continuous inflammation in HBeAg-negative patients coinfected with HCV. Gastroenterol Jpn 1992;27:617-623. Key words:
anti-CP-9; anti-HCV; coinfection of HBV and HCV; H B V carrier; HCV-RNA.
Introduction Development of hepatitis C virus (HCV) infection diagnosis kits revealed that the major cause of non-A, non-B (NANB) hepatitis is H C V infection ~,2. In NANB hepatitis, 70% of patients react to anti-cl00-3, which acts against proteins encoded by non-structured proteins (NS3 and NS4 region) 1'2, but anti-cl00-3 was also detected in 10% of patients with type B chronic liver disease 3-9. By detecting anti-cl00-3 in type B chronic liver disease, Fattovich et al8 reported that H C V might play a leading role in causing liver diseases with antibody to HBe antigen (anti-HBe)-positive among HBV-DNA-polymerase (DNA-P)-negative patients. Fong et al9 reported H C V appeared to suppress hepatitis B virus (HBV) replication and
to cause severe liver diseases in patients with anticl00-3-positive type B chronic liver disease. However, in these papers, past infection of H C V was not completely studied because HCV-RNA was not estimated in anti-cl00-3 positive patients. To identify present coinfection with HBV and HCV, we estimated both HCV-RNA by reverse transcription polymerase chain reaction (RTPCR) 1~ and antibody against a core protein of H C V (anti-CP-9) by enzyme immunoassay (EIA) 12,13 in anti-cl00-3 positive type B chronic liver disease. Furthermore, to assess the difference between coinfected liver disease and type B chronic liver disease or type C chronic liver disease, we evaluated the viral markers of HBV and HCV, clinical data, and histological features in these patients.
Received November 27, 1991. Accepted May 22, 1992. Address for correspondence: Toshihiko Doi, M.D., The First Department of Internal Medicine, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700, Japan.
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T. Doi et aL
Vol. 27, No. 5
Table 1. Positivity of anti-c100-3 antibody in 152 patients with HBsAg-positive chronic liver disease categorized according to clinical diagnosis of liver disease Clinical diagnosis
No. of patients
No. of anti-cl00-3-positive (%)
Chronic hepatitis Liver cirrhosis without HCC Liver cirrhosis with HCC
95 37 20
4(4.2) 5(13.5) 2(10.0)
Total
152
11 (7.2)
HCC: hepatocellular carcinoma.
Table 2. Positivity of anti-c100-3 antibody in subgroups of chronic hepatitis B HBV markers
No. of patients
HBeAg-positive HBeAg-negative HBV-DNA-P positive HBV-DNA-P negative Not determined
93 59 6 44 9
Subjects and Methods One hundred fifty-two patients with HBs antigen (HBsAg)-positive chronic liver disease were studied. Ninety-five patients were diagnosed as chronic hepatitis by liver histology and 57 patients as cirrhosis, with or without hepatocellular carcinoma (HCC), by liver histology and/or by imaging examinations such as ultrasonography, computed tomography, and angiography (Table 1). Sera of patients stored at -80~ were examined. As for HCV-associated antibodies, the antibody against NS3 & 4 region proteins (anti-cl00-3) was tested by EIA (Ortho Co., NJ, USA) u and the antibody against the core protein of HCV (antiCP-9) was tested by EIA (Okamoto et al) 12']3. The results of anti-cl00-3 and anti-CP-9 were expressed as optical density (OD) (OD positive >0.450). HCV-RNA was tested using RT-PCR 1~ As its primer, 5'non-coding region of HCV-RNA was applied ~. HBe antigen (HBeAg) and anti-HBe were measured by radioimmunoassay (Abbott laboratories, IL, USA), and HBV-DNA-P was determined by Kaplan's method ]4. Liver histology of 9 coinfected patients was evaluated using both Mallory-Azan and H&E stains. Periportal piecemeal necrosis, intralobular degen-
No. of anti-cl00-3-positive (%) 4(4.3) 7(11.9) 0 7 0
eration and portal inflammation were scored according to Knodell's HAI score ]5.
Statistical analysis Statistical analysis was carried out using the chisquare test or the two-tailed Student t-test when samples were of sufficient size. A two-tailed P value of
Gastroenterologia Japonica Copyright 9 1992 by The Japanese Society of Gastroenterology
Hepatitis type C virus infection in patients with type B chronic liver disease Toshihiko DOI, Gotaro YAMADA, Hisashi ENDO, Hiroshi NISHIMOTO, Michiko TAKAHASHI, Rieko MIYAMOTO, Shigeatsu FUJIKI, Hiroyuki SHIMOMURA, Motowo MIZUNO, and Takao TSUJI First Department of Internal Medicine, Okayama University Medical School Okayama, Japan Summary: Anti-cl00-3 (Ortho) was determined in the sera of 152 patients with HBs antigen-positive chronic liver diseases to assess coinfection of hepatitis B virus (HBV) and hepatitis C virus (HCV). Eleven patients (7.2%) were positive for anti-cl00-3. Anti-CP-9 (Okamoto) and HCV-RNA (RT-PCR) were also examined in these 11 patients. Anti-CP-9 was detected in 7 patients and HCV-RNA was detected in all 11 patients. Four of the 11 anti-cl00-3-positive patients were positive for HBe antigen (HBeAg) and others were negative. In 8 of the 11 patients, HCV was suspected to be superinfected by blood transfusion. In HBeAgpositive patients, serum glutamic pyruvic transaminase (SGPT) was elevated in relation to active replication of HBV shown by DNA-polymerase activity. The histological findings showed chronic active hepatitis, with or without cirrhosis. On the other hand, in HBeAg-negative patients, SGPT fluctuated without evidence of active replication of HBV. Active inflammation in the liver was observed in 3 of 5 HBeAg-negative patients by liver biopsy. These findings suggest that HBV might play an important role in chronic active inflammation in HBeAg-positive patients coinfected with HCV, and that HCV might be responsible for continuous inflammation in HBeAg-negative patients coinfected with HCV. Gastroenterol Jpn 1992;27:617-623. Key words:
anti-CP-9; anti-HCV; coinfection of HBV and HCV; H B V carrier; HCV-RNA.
Introduction Development of hepatitis C virus (HCV) infection diagnosis kits revealed that the major cause of non-A, non-B (NANB) hepatitis is H C V infection ~,2. In NANB hepatitis, 70% of patients react to anti-cl00-3, which acts against proteins encoded by non-structured proteins (NS3 and NS4 region) 1'2, but anti-cl00-3 was also detected in 10% of patients with type B chronic liver disease 3-9. By detecting anti-cl00-3 in type B chronic liver disease, Fattovich et al8 reported that H C V might play a leading role in causing liver diseases with antibody to HBe antigen (anti-HBe)-positive among HBV-DNA-polymerase (DNA-P)-negative patients. Fong et al9 reported H C V appeared to suppress hepatitis B virus (HBV) replication and
to cause severe liver diseases in patients with anticl00-3-positive type B chronic liver disease. However, in these papers, past infection of H C V was not completely studied because HCV-RNA was not estimated in anti-cl00-3 positive patients. To identify present coinfection with HBV and HCV, we estimated both HCV-RNA by reverse transcription polymerase chain reaction (RTPCR) 1~ and antibody against a core protein of H C V (anti-CP-9) by enzyme immunoassay (EIA) 12,13 in anti-cl00-3 positive type B chronic liver disease. Furthermore, to assess the difference between coinfected liver disease and type B chronic liver disease or type C chronic liver disease, we evaluated the viral markers of HBV and HCV, clinical data, and histological features in these patients.
Received November 27, 1991. Accepted May 22, 1992. Address for correspondence: Toshihiko Doi, M.D., The First Department of Internal Medicine, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700, Japan.
618
T. Doi et aL
Vol. 27, No. 5
Table 1. Positivity of anti-c100-3 antibody in 152 patients with HBsAg-positive chronic liver disease categorized according to clinical diagnosis of liver disease Clinical diagnosis
No. of patients
No. of anti-cl00-3-positive (%)
Chronic hepatitis Liver cirrhosis without HCC Liver cirrhosis with HCC
95 37 20
4(4.2) 5(13.5) 2(10.0)
Total
152
11 (7.2)
HCC: hepatocellular carcinoma.
Table 2. Positivity of anti-c100-3 antibody in subgroups of chronic hepatitis B HBV markers
No. of patients
HBeAg-positive HBeAg-negative HBV-DNA-P positive HBV-DNA-P negative Not determined
93 59 6 44 9
Subjects and Methods One hundred fifty-two patients with HBs antigen (HBsAg)-positive chronic liver disease were studied. Ninety-five patients were diagnosed as chronic hepatitis by liver histology and 57 patients as cirrhosis, with or without hepatocellular carcinoma (HCC), by liver histology and/or by imaging examinations such as ultrasonography, computed tomography, and angiography (Table 1). Sera of patients stored at -80~ were examined. As for HCV-associated antibodies, the antibody against NS3 & 4 region proteins (anti-cl00-3) was tested by EIA (Ortho Co., NJ, USA) u and the antibody against the core protein of HCV (antiCP-9) was tested by EIA (Okamoto et al) 12']3. The results of anti-cl00-3 and anti-CP-9 were expressed as optical density (OD) (OD positive >0.450). HCV-RNA was tested using RT-PCR 1~ As its primer, 5'non-coding region of HCV-RNA was applied ~. HBe antigen (HBeAg) and anti-HBe were measured by radioimmunoassay (Abbott laboratories, IL, USA), and HBV-DNA-P was determined by Kaplan's method ]4. Liver histology of 9 coinfected patients was evaluated using both Mallory-Azan and H&E stains. Periportal piecemeal necrosis, intralobular degen-
No. of anti-cl00-3-positive (%) 4(4.3) 7(11.9) 0 7 0
eration and portal inflammation were scored according to Knodell's HAI score ]5.
Statistical analysis Statistical analysis was carried out using the chisquare test or the two-tailed Student t-test when samples were of sufficient size. A two-tailed P value of
