Kohachiro Sugiyama

Transmission of hepatitis B virus from hepatitis B core antibody- positive donors in living related liver transplants

BACKGROUND In order to clarify the risk of hepatitis B virus (HBV) transmission from hepatitis B core antibody-positive (HBcAb(+)) donors and to evolve a new strategy to counter such a risk, we undertook a retrospective (1990-1995) and prospective (1995-1996) analysis of our experience with living related liver transplantation involving HBcAb(+) donors. METHODS Between June 15, 1990, and June 30, 1995, HBcAb(+) individuals were not excluded as donor candidates at our institutions. For 171 liver transplants, 16 donors were HBcAb(+). Between July 1, 1995, and June 30, 1996, HBcAb(+) individuals were generally excluded as donor candidates; however, three recipients were given liver grafts from HBcAb(+) donors because other donor candidates presented even higher risks. In the latter period, recipients with transplants from HBcAb(+) donors underwent prophylactic passive immunization with hyperimmune hepatitis B immunoglobulin (HBIG). The serum of 10 HBcAb(+) donors was examined by nested polymerase chain reaction for the presence of HBV-DNA, but it was not detected in any of them. However, the same examination of the liver tissue of five such donors yielded positive results in all cases. RESULTS In the first 5-year period, out of 16 recipients with HBcAb(+) donors, 15 became hepatitis B surface antigen-positive after transplant. The three recipients with HBcAb(+) donors during the second 1-year period, who were treated by prophylactic passive immunization with HBIG, remained hepatitis B surface antigen-negative and negative for serum HBV-DNA after transplant. CONCLUSIONS HBV exists in the liver of healthy HBcAb(+) individuals, but not in the blood. Therefore, HBV is thought to be transmitted to recipients by liver grafts from the HBcAb(+) donors at a significantly high rate. The prevention of viral activation and clinical disease development by means of passive immunization with HBIG seems promising, although the follow-up period in our study may be too short for any definitive conclusions.

A pediatric patient with classical citrullinemia who underwent living-related partial liver transplantation.

Patients with inborn errors of metabolism undergo liver transplantation, but the effect of transplanting the liver of healthy carriers of these conditions has not been fully clarified. A 6-year-old girl with classical citrullinemia, who repeatedly suffered from hyperammonemia, underwent living-related liver transplantation by using a segment of the liver of her mother, who was a heterozygote carrier for classical citrullinemia. Hyperammonemia alleviated in the patient after the transplantation, thereby dramatically improving her quality of life. Although the levels of plasma and urinary citrulline remained high postoperatively, there was no marked difference in the level of plasma citrulline up to 1 month after surgery when compared with that of previously reported orthotopic liver transplantation cases with classical citrullinemia.

Bile salt export pump gene mutations in two Japanese patients with progressive familial intrahepatic cholestasis.

BackgroundIn recent years, progressive familial intrahepatic cholestasis has been classified into at least three types by genetic analysis: PFIC1, PFIC2, and MDR3. Liver transplantation is effective for treating patients with this intractable syndrome. Confirming the correct diagnosis is of paramount importance because prognosis after transplantation differs with the genetic type of the disease. MethodsSynthesis of cDNA was accomplished using RNA extracted from liver tissue of two Japanese patients with progressive familial intrahepatic cholestasis. Polymerase chain reaction was performed using 13 primer sets designed for amplification of the bile salt export pump cDNA. Direct sequencing was undertaken, and identified sequences were compared with the sequence for bile salt export pump gene registered with GenBank. In addition, gene sequences for nonprogressive familial intrahepatic cholestasis patients were analyzed. ResultsGenetic analysis of patient 1 revealed that substitutions in bile salt export pump protein sequences, namely R575X and E636G, might be the cause of the disease. In patient 2, V330X and R487H might fulfill the same role. Results of gene analysis in parents and cholestatic controls supported these conclusions. ConclusionsAbsence or presence of bile salt export protein gene mutations was confirmed as representing a useful prognostic marker for clinical course after liver transplantation.

Route of TT virus infection in children

TT virus (TTV) is a novel viral agent, detected recently in non‐A to E hepatitis cases. Little is known about its natural history or routes of transmission in childhood. For the detection of serum TTV DNA, semi‐nested polymerase chain reaction (PCR) was carried out using TTV‐specific primers and TTV nucleotide sequences were determined by the dideoxy chain‐mediated termination method. Five of the 70 children studied (including 20 hepatitis B virus [HBV] carriers, 40 children born to HBV carrier mothers and 10 children born to hepatitis C virus [HCV] carrier mothers) had serum TTV DNA. Three of the 5 children had siblings (4 in total), so that a total of 9 children were studied to determine the time of initial serum TTV DNA detection. In the 8 seropositive children, the time of serum TTV DNA detection ranged from 6 to 14 months after birth, and TTV DNA persisted thereafter throughout the follow‐up period. The TTV DNA‐negative child was assessed most recently at 6 months of age. TTV DNA was detected in only 2 of the 4 mothers tested (families 2 and 3). When 271‐bp TTV DNA fragments from each of the 8 children were sequenced, the degree of homology between siblings in families 1–3 was 100%, 99.5%, and 92.3%, respectively. The degree of homology between child‐mother pairs of families 2 and 3 was 99.5–100% and 62.6–63.9%, respectively. The distribution of different TTV strains was consistent within families, except for family 3. None of the TTV‐infected children had elevated levels of alanine aminotransferase or clinical signs of liver disease. J. Med. Virol. 59:204–207, 1999.

Detection rates of TT virus among children who visited a general hospital in Japan

Recently, genomic DNA of the novel TT virus (TTV) was isolated from patients suffering from posttransfusion hepatitis of unknown etiology. We examined sera from 197 children who visited the Department of Pediatrics at Toyohashi National Hospital. Sera were tested for TTV DNA by seminested polymerase chain reaction (PCR) using a set of primers synthesized according to the published TTV sequence. Ten children were found to be positive for TTV (5.1%). All positive PCR products were directly sequenced in both directions using a fluorescent dye terminator cycle sequencing system. The sequences were compared by a multiple sequence alignment and a phylogenetic tree was constructed. The phylogenetic tree showed that two of the TTV isolates found in the present experiment did not belong to any of the phylogenetic groups previously reported. J. Med. Virol. 57:405–407, 1999.

Antley-Bixler syndrome in a sister and brother

SummaryA sister and brother both with the clinical and radiographic features of Antley-Bixler syndrome are reported. Their parents were first cousins. Analysis of these siblings and seven other patients reported in the literature indicates autosomal recessive inheritance for the syndrome.

Prevalence of TTV DNA among children with a history of transfusion or liver disease.

The prevalence rates of serum TT virus (TTV) DNA among children with or without a history of transfusion or liver disease were studied by polymerase chain reaction (PCR) using either the Okamoto primer set or the Takahashi primer set developed more recently. Using Okamoto and Takahashi primer sets, the prevalence rates were 31.6% (12/38) and 78.9% (30/38), respectively, for children with a history of blood transfusion (including malignant and non‐malignant groups) and 6.7% (2/30) and 60% (18/30), respectively, for children without a history of blood transfusion. Among pregnant women, these rates were 12.9% (4/31) and 61.3% (19/31), respectively. On the other hand, the prevalence rates were 0% (0/16) and 50% (8/16), respectively, in hepatitis B patients, 21.4% (3/14) and 71.4% (10/14), respectively, for hepatitis C patients, and 20.0% (9/45) and 57.8% (26/45), respectively, for non‐A to C hepatitis patients (including 27 acute hepatitis patients, 5 fulminant patients and 13 chronic hepatitis patients). In this study, the prevalence rates determined by the Takahashi primer set tended to be 2–9 times higher than those determined using the Okamoto primer set. These results suggest that TTV infection is widespread among Japanese children. Furthermore, blood transfusion does not appear to be the major route of infection. The similar prevalence rates between control children and children with various types of hepatitis using the Takahashi primer system suggest that TTV infection does not play a direct causative role in the development of liver disease in children. J. Med. Virol. 60:172–176, 2000.

The first case of the Antley-Bixler syndrome with a consanguinity in Japan

SummaryAntley-Bixler syndrome has the specific features as brachycephaly, craniosynostosis, midface hypoplasia, depressed nasal bridge, proptosis, dysplastic ears, upper respiratory obstruction, radiohumeral synostosis, joint contructures, arachnodactyly, and femoral bowing and fractures. It is a very rare disease and only five cases have been reported. Its etiology and heredity are unknown. All reported cases were sporadic but present case has the cousinship marriage parents, suggesting genetic disorder of autosomal recessive type.

Highly diverse TTV population in infants and their mothers.

Infants born to serum HCV-positive 12 mothers were enrolled in the study. Nucleotide sequences amplified by primers deduced from a noncoding region were compared between mothers and their infants. The rates for detection of serum TTV in 12 mothers and their infants were 10/12 (83%) and 9/12 (75%), respectively. Serum TTV DNA was not detected in any infant at 1 month of age, but was detected for the first time between 1.5 and 8 months after birth. Positivity persisted thereafter throughout the follow-up period. In seven randomly selected mother-infant pairs, intrahost TTV heterogeneity was lower in infants than in mothers. Furthermore, one of seven mother-infant pairs showed a high degree of similarity (98.7-100%) in all clones, while in four infants, all nucleotide sequences differed by >10% from those of their mothers. However, the degree of homology in the two mother-infant pairs was 89-98.7% in family 2 and 88.1-99.4% in family 5. In the present study, with only one exception, it was shown that TTV from infants is not identical to TTV from mothers. The mechanism is discussed briefly in this paper.

Prevalence of hepatitis E virus infection in Japanese children

Background: Recently, sporadic cases of acute hepatitis and fulminant hepatitis caused by hepatitis E virus (HEV) have been reported in Japan. However, few reports have addressed the issue of HEV infection during childhood. Methods: This study included 5 patients with fulminant hepatitis, 30 patients with acute hepatitis, and 309 patients without history of hepatic dysfunction or hepatitis in childhood as control. RNA was extracted from each serum sample, and HEV specific reverse-transcriptase polymerase chain reaction was performed. Anti-HEV immunoglobulin (Ig)M and IgG were measured by enzyme-linked immunoadsorbent assay. Results: HEV RNA, anti-HEV IgM, and anti-HEV IgG were not detected in the sera of any of the five patients with fulminant hepatitis. In the 30 patients with acute hepatitis, only one (3.3%) was positive for anti-HEV IgG, and all were negative for anti-HEV IgM and HEV RNA. Of the 309 control patients, 8 (2.6%) were positive for anti-HEV IgG, and 2 (0.6%) were positive for anti-HEV IgM, respectively. Conclusion: The result of patients with fulminant hepatitis suggests that HEV is an unlikely cause of fulminant hepatitis in children. However, the detection rate of anti-HEV IgG shows that a history of HEV infection is not so rare among children in Japan.