Kaori Fujii

Determination of the nucleotide sequences of heat shock operon groESL and the citrate synthase gene (gltA) of Anaplasma (Ehrlichia) platys for phylogenetic and diagnostic studies.

ABSTRACT The 1,670-bp nucleotide sequence of the heat shock operon groESL and the 1,236-bp sequence of the citrate synthase gene (gltA) of Anaplasma (Ehrlichia) platys were determined. The topology of the groEL- and gltA-based phylogenetic tree was similar to that derived from 16S rRNA gene analyses with distances. Both groESL- and gltA-based PCRs specific to A. platys were also developed based upon the alignment data.

Demonstration of Anaplasma (Ehrlichia) platys inclusions in peripheral blood platelets of a dog in Japan.

A free-roaming dog in Okinawa island showed Anaplasma (Ehrlichia) platys-like inclusions within the platelets of peripheral blood samples. The inclusions were positive in indirect fluorescence test with anti-A. phagocytophila serum. The platelet count of the dog was 170,000 microl(-1). The sequence analysis of the 16S rRNA, citrate synthase and heat shock protein genes of DNA from the infected platelets confirmed that the inclusions were A. platys. This is the first detection of A. platys inclusions in dogs in Japan.

Hepatic Fas antigen expression before and after interferon therapy in patients with chronic hepatitis C.

To assess the relationship between hepatitis C virus infection and Fas antigen expression on hepatocytes, we examined changes in hepatic Fas antigen expression in the presence or absence of active hepatitis C virus infection. Twenty patients with chronic hepatitis C infection were treated with interferon and underwent pre- and posttreatment liver biopsies. Patients were classified according to the absence (group A; n=9) or the presence (group B; n=11) of hepatitis C virus RNA (HCV-RNA) in the liver after interferon therapy. An immunohistochemical assay showed Fas antigen staining in hepatocytes membranes and cytoplasm with expression concentrated mainly in periportal areas. The percentage of Fas-positive cells in the liver before treatment was not different between group A (39.5 ± 19.1%) and group B (32.5 ± 15.6%). Hepatic Fas expression was reduced significantly after treatment (24.3 ± 10.6%) compared with the pretreatment values in group A (p < 0.05) but not in group B (25.9 ± 16.9%). There was no significant difference between the two groups in the degree of histologic improvement. These results suggest that hepatic Fas expression is associated with persistent infection of hepatitis C virus.

Glycine-to-arginine substitution at codon 145 of HBsAg in two infants Born to hepatitis B e antigen-positive carrier

Two daughters born to an HBeAg-positive carrier became hepatitis B virus carriers despite immunoprophylaxis with hepatitis B immune globulin or hepatitis B vaccine. The elder daughter received hepatitis B immune globulin alone and the younger daughter received both hepatitis B immune globulin and hepatitis B vaccine. They acquired anti-HBs passively immediately after birth, and the younger daughter did not respond to active immunization. The nucleotide sequence of the S gene of hepatitis B virus DNA from the four carriers in the family was determined. A 531-bp fragment within the S gene, which includes a region encoding both the commona and type-specificd/y orr/w determinants, was amplified by polymerase chain reaction and sequenced. Antigenic subtypes of HBV were identified asadr in the father andadw in both the mother and two daughters. Amino acid residues 122–160 of HBsAg were identical between the daughters and their mother except for a glycine-to-arginine substitution at codon 145. A possibility that this escape mutant had selective advantage over wild-type hepatitis B virus under immune pressure is discussed.

Immunohistochemical study of Fas antigen in liver of patients with chronic hepatitis and autoimmune liver disease

Abstract To examine the presence of Fas antigen in the liver in various chronic liver diseases, we stained Fas antigen immunohistochemically in the livers of 12 patients who were diagnosed as chronic hepatitis C in five, chronic hepatitis B in three, primary biliary cirrhosis in three, and autoimmune hepatitis in one. Three specimens with non-specific reactive hepatitis (NSRH) were used as histological controls. We prepared two kinds of rabbit polyclonal antibodies (anti-C, anti-D) as the primary antibody, raised against synthetic polypeptides from the two regions of human Fas antigen (C region, aa 300–314; D region, aa 104–114). The degree of the presence of Fas antigen expressed as the mean labeling index was relatively higher in chronic liver disease than in NSRH. Fas antigen was stained in the cytoplasm regardless of differences in primary antibody, but more diffusely in antibody C. Fas antigen was randomly distributed in the lobule and mainly concentrated in the periportal area associated with piecemeal necrosis. These results suggest that Fas antigen is related to liver injury from chronic viral hepatitis and autoimmune liver disease.

Correlation Between Relative Number of Circulating Low-Density Hepatitis C Virus Particles and Disease Activity in Patients with Chronic Hepatitis C

Hepatitis C virus (HCV) circulates as particleshaving differing buoyant densities. Changes in therelative proportions of virus particles of differentdensities were examined in 19 patients with chronic hepatitis C: 6 without (group A) and 13 with(group B) abnormal serum alanine aminotransferase (ALT)levels. High- and low-density virus particles wereseparated by differential flotation centrifugation. The numbers of high-density particlesconsistently exceeded that of low-density particles inall patients in group A, whereas the titers of bothtypes of particles were the same at least once in 7 of10 patients sampled at two time points in group B.The ALT level significantly increased, <2 monthslater (P < 0.05) when the titers of both types ofparticles were the same in patients in group B. Thus, we found a correlation between the relativenumbers of circulating low-density HCV particles anddisease activity in chronic hepatitis Cpatients.

Clinical evaluation of chronic liver diseases induced by different hepatitis C virus genotype in Japan, focused on the difference between genotypes III/2a and IV/2b

It remains controversial if hepatitis C virus (HCV) genotypes in the same genetic group or type are different in the activity of viral replication and capacity to induce severe clinical disease. HCV genotypes and HCV RNA titers were determined in consecutive 264 patients with chronic liver disease in Yamaguchi, Japan. Genotype I/1a was detected in 3 (1%), II/1b in 192 (74%), III/2a in 46 (18%), IV/2b in 17 (6%) and co-infection with II/1b and III/2a in 3 (1%); HCV RNA titers in the remaining three patients were too low to be genotyped. The patients with genotype III/2a were significantly older (P<0.05) and had lower HCV RNA titers in serum than those with genotype IV/2b (P=0.0211; odds ratio 6.47 [95% confidence interval: 1.323–31.587]). These results indicate that, even though genotypes III/2a and IV/2b belong to the same genetic group (type) 2, they would need to be distinguished clinically because the patients’ age and HCV RNA titer in serum are both important factors in treating the patients with HCV infection.

Rebound phenomenon of hepatitis C viremia after interferon therapy in "relapsed" patients with chronic hepatitis C

The correlation between serum hepatitis C virus (HCV)-RNA and aminotransferase levels following completion of interferon therapy was evaluated in eight relapsed patients with chronic hepatitis C. Both HCV-RNA and aminotransferase levels were significantly increased in the relapsed patients 1 month after completion of therapy, compared to pretreatment values, despite aminotransferase levels being normal and HCV-RNA being undetectable by reverse transcription polymerase chain reaction assay at the end of therapy. The serum levels of HCV-RNA and aminotransferase were significantly elevated 1 and 2 months post-treatment. They then decreased to pre-treatment values 3–5 months after the completion of therapy. Thus, in relapsed patients after the completion of therapy, the changes in HCV-RNA levels preceded the elevation in aminotransferase levels. These findings suggest a correlation between serum HCV-RNA levels and aminotransferase levels in relapsed patients with chronic hepatitis C after the completion of interferon therapy.