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416
was determined by a quantitative method (Genostics Tm Abbott) and by PCR in S and C regions. Results are presented in Table 1. In anti-HCV(+) Ab patients, 4/19 (21%) were HCV-RNA-PCR positive and had an alcoholic cirrhosis. All four had C33-c- and C223-positive bands, but ClOO-3 and 5-l-l bands were present in one and two patients, respectively. The only HBV marker was anti-HBc Ab in two patients. In anti HCV(-) Ab patients, l/21 (4.8%) was HCV-RNA-PCR positive, with a post-hepatitic cirrhosis. No I-IBV serum marker was detected excepted for anti-HBc Ab. HBVDNA was negative by Genostics and PCR. Four of five HCV-RNA-PCR( +) patients were native from South European countries. In conclusion, our results suggest that the presence of HCV-RNA in the serum of Caucasian patients with HCC is uncommon. It is noteworthy that, as in an other study (6) C33c and C22-3 bands seemed to be related to infectivity. The geographic origin of patients (mainly from North European countries) and the etiology of cirrhosis (mainly alcoholic) could account for the low prevalence of the HCV genome. Another hypothesis for the low level of HCV in serum might be the absence of vire,mia at this stage of the disease. DNA
TABLE 1 Characteristics
TO THE EDITOR
of HCC patients acccnding to anti-HCV status Anti-HCV( +) (n = 19)
Riba II(+) ClOO-3 S-!-l cz-3 c33-c HCV-RNA-PCR(
+)
9 (47%) 10 (53%) 16 (84%) 15 (79%) 4 (21%)
Anti-HCV(-) (n = 21)
1 (4.8%)
cirrhosis (alcoholic, n = 24; post-hepatitic, n = 9; mixed, n = 6; indeterminate, n = 1). The mean age (67 and 69.25 year), geographic origin (six South Europeans119 (31.6%) and six South Europeans/21 (28.6%)) and sex distribution (male to female ratio = 14/5 (2.8:1) and 1615 (3.2:1), respectively, were not different in the two groups. HCV Ab were sought by ELISA II and RIBA II (Qrtho diagnostic systems, Raritan, NJ, U.S.A.) in which two HCV recombinant antigens were added: one in a non-structural region (C33-c) and the other one in an HCV-core associated antigen (C22-3). Sera reacting with two or more of the four RIBA antigens were considered positive, those reacting with one antigen were indeterminate, and sera that did not react with any antigen were negative. The presence of HCV-RNA in sera from patients was investigated using nested PCR. Two sets of nested oligonucleotide primers based on the Chiron sequence and located in the highly conserved 5’ non-coding region (5) were used. Sera were also tested for HBV markers by ELISA (Abbott, Chicago, U.S.A.). HBVReferences 1 Bruix J, Barrera JM, Calvet X, et al. Prevalence of antibodids to hepatitis C virus in Spanish patients with hepatocellular car& noma and hepatic cirrhosis. Lancet 1989; ii: 1004-6. 2 Simonetti RG, Cottone M, Craxi A, et al. Prevalence of antibodies to hepatitis C virus in hepatocellular carcinoma. Lancet 1990: ii:1338. 3 Nalpas B, Driss F, Pol S, Hamelin B, Brtchot C, Berthelot P.
M.A. Thelu’,*, J.P. Zarsk?*, and J.M. Seigneurin2
D. Dardelet’**, F. Lune13
‘Clinique d’H@pato-Gastroenthologie, ‘Laboratoire de Viroiogie, CHRU BP 217 38043, Grenoble and 3Lnboratoire de Virologie, Hbpital Salp&i&e, Paris, France
Association between HCV and HBV infection in hepatocellular carcinoma and alcoholic liver disease. J Hepatol 1991; 12: 60-j. Zarski JP, Lunel F, Dardelet D. et al. Hepatitis C virus a.nd hepatocellular carcinoma in France: detection of antibodies using two second generation assays. J Hepatol 1991; 13: 376. Garson JA, Tedder RS, Briggs M, et al. Detection of hepatitis C viral sequences in blood donations by ‘nested’ polymerase chain reaction and prediction of infectivity. Lancet 1990; 335: 1419-22. Alberti A. Diagnosis of hepat.iis C. J Hepatol 1991; 12: 279-82.
HEPAT 01046
Prevalence of hepatitis C virus among non-A, nonliver disease in Egypt Until recently, hepatitis due to NANB agent (1) has been a diagr.cjsis of serological exclusior!. The development of a recombinant antigen-based immunoassay for HCV circulating antibodies (2) has provided a reliable
-related chronic
serological marker for chronic HCV infection. II: was our aim to report for the first time from Egypt the Yrevalence of anti-HCV in patients diagnosed by serological exclusion as non-A, non-B chronic liver dis-
LETTERS
TO THE EDITOR
417
TABLE 1 Age, se?:, anti-HCV prevalence related chronic liver disease
---~~ No.
_~_____
in healthy control ___Controls
Male (8)
76 100%
Mean age (yr) Anti-HCV +ve (%) Statistical significance
43p5.2, p < 0.6001
ani
NANB-
NANB-CLD 160 80.6% 41 107 (66.8)
TABLE 2 Risk factor of NANB hepatitis among anti-HCV-positive and antiHCSI-negative patients --~___~ --. Transmission Anti-HCV Anti-HCV p-value -ve +ve (n = 107) (n = 53) Transfusion-related Nosocomial-acquired Community-squired
28 (26.2%) 54 (50.4%) 2.5 (23.4%)
6 (11.3%) 29 (54.7%) 8 (34.0%)
< 0.05 N.S. N.S.
ease (NANB-CLD) and to assess the possible risk factors of infection among these patients. Scra of 160 Egyptian patients, 129 males and 31 females with a mean age of 41 years diagnosed by serological exclusion as NANB-CLD and sera of 76 volunteer blood donors (controls) were tested repeatedly for antiReferences 1 Esteban JI. Esteban R, Viladomin L, et al. Hepa,;ir.is C virus antibodies among risk groups in Spain. Lancet 1989; ii: 294-6.
HCV using Ortho-diagnostic system Inc. kits. Anti-HCV was reactive in 66.8% of 160 NANB-CLD patients compared to 5.2% of 76 controls @ < 0.001) as shown in Table 1. Epidemiological interview about modes of transmission among anti-HCV-positive patients (n = 107), revealed that 26.2% were transfusion-related (TR), 50.4% nosocomial-acquired (NA) and 23.4% presumed community-acquired (CA). Although transfusion was the only significant risk factor, it contributed to a small proportion (26.2%) of transmission of HCV compared to non-transfusion routes (Table 2). Anti-HBc was significantly higher in the anti-HCVpositive cases (60.7%). compared to the anti-HCVnegative (39.6%) group @ < 0.05). This is explained by a common route of transmission of both viruses. Histological examination of the biopsy specimens in 66 antiHCV-positive and 50 anti-HCV-negative patients showed that cirrhosis was significantly increased (24.5%) in the anti-HCV-positive patients (p K O.Ol), whereas chronic persistent hepatitis (CPH) was significantly high (36.0%) in the anti-HCV-negative patients @ < 0.01).
A. Ei-Zayadi, El-Haddad Fan&y
Osaima Selim, Mona Rafik and Salwa
of Medicine.
Ain-Shams
University, Cairo. Egypt
2 Kuo 6, Choo AL, Alter HJ. et al. An assay of circulating antibodies to a major ctiologic virus of hum; I non-A, non-B hepatitis. Science 1989; 244: 362.
HEPAT 01051
centrat~ons increase wit lications after liver trans Soluble class I antigens have been detected in the serum of patients after liver transplantation (1). The grafted liver delivers soluble class I antigens into the recipient’s circulation, although class 1 antigens are also derived from other sources (1). We used a recently developed enzyme-linked immunosorbent assay (2) to investigate serum concentrations of sHLA class I molecules during transplant-related complications following liver transplantation. Sixteen patients (12 male and four female) after orthotopic liver transplantation were studied (mean age, 42.5 years). For prophylactic immunosuppression cyclo-
spot-me A, azathioprine and steroids were used. Rejection episodes were treated with gram doses of methylprednisone. The serum samples tested were stored below --20°C for a maximum of 1 year. &Microglobulin (B,M) was assayed by a commercially available radioimmunoassay (Pharmacia Diagnostics). TNF-a and IFN-y were analyzed bv ELISAs (IRE Medgenix). Rejection and CMV disease were diagnosed as reported recently (3). Wilcoxon’s matched pairs signed-rank test and Pearson correlation test were used for statistical analysis. A total of 252 serum samples from sixteen liver transplant patients were analyzed. Six patients showed an