Toshio Kawatani

Incidence of hepatitis virus infection and severe liver dysfunction in patients receiving chemotherapy for hematologic malignancies

Abstract: Hepatitis virus infection through virus reactivation has a high risk of mortality in patients with hematological malignancies receiving chemotherapy. We examined the incidence of both hepatitis B virus (HBV) and hepatitis C virus (HCV) infection and severe liver dysfunction (alanine aminotransferase >ten times the normal upper limit and total bilirubin >5 mg/dl) during chemotherapy in 268 patients with hematological malignancies. Eight patients (3.0%) were infected with HBV and 22 patients (8.2%) were infected with HCV. One patient (0.4%) was infected with both HBV and HCV. HBV‐ or HCV‐infected patients showed severe liver dysfunction at a significantly higher incidence than non‐infected patients (11/31 (35.5%) vs. 0/237 (0%), p<0.0001). Furthermore, the incidence of severe liver dysfunction in HBV‐infected patients was significantly higher than in HCV‐infected patients (6/8 (75.0%) vs. 4/22 (18.2%), p<0.01). Three of eight HBV‐infected patients were initially negative for hepatitis B surface antigen (HBsAg) by latex agglutination and became positive for HBsAg during chemotherapy. Furthermore, all three patients developed severe liver dysfunction and two developed fatal fulminant hepatitis. From an examination of the original stock of serum samples before chemotherapy, two patients were found to be positive for HBV‐DNA by polymerase chain reaction (PCR). Although post‐transfusion HBV infection was suspected in the one remaining patient, the cause of HBV infection could not be clarified due to the impossibility of examination in blood donors. Since HBV‐infected patients develop severe liver dysfunction at a higher incidence than either patients not infected with virus or HCV‐infected patients before chemotherapy for hematological malignancies, it is recommended that HBV‐DNA should be tested by PCR to detect HBV marker‐negative carriers and liver function tests should be carefully monitored.

Serum soluble c‐kit receptor and expression of c‐kit protein and mRNA in acute myeloid leukemia

Abstract: To investigate the clinical role of the soluble form of c‐kit receptor (s‐kit) in patients with acute myeloid leukemia (AML), we determined the levels of serum s‐kit and expression of c‐kit antigens and mRNA in leukemic cells. The serum s‐kit level was measured using ELISA assay in 30 AML patients and 20 normal controls. C‐kit antigens of leukemic blasts were stained immunohistologically, and c‐kit mRNA was detected by RT‐PCR. The serum s‐kit level in M1 and M2 were significantly increased (p<0.01) and that in M4 or M5 was significantly decreased (p<0.05) compared to that in the controls. In the comparisons among subtypes of FAB classification, M1 and M2 showed significantly higher levels than M4 or M5 (p<0.05 and p<0.01, respectively). Both expression of c‐kit antigens and mRNA were observed in M0 (1/4), M1 (2/4) and M2 (6/8), but neither was observed in M4 or M5. The serum s‐kit levels were correlated with the absolute number of AML blasts in peripheral blood (r=0.564, p<0.05). These results indicate that the serum s‐kit level is related to the stage of differentiation of AML blasts in accordance with the expression of c‐kit protein and mRNA in AML blasts, and is useful for assessment of leukemic cell burden.

Fulminant hepatitis type B after chemotherapy in a serologically negative hepatitis B virus carrier with acute myelogenous leukemia.

We report a case of a 41-year-old man with acute myelogenous leukemia who developed fulminant hepatitis from reactivation of trace hepatitis B virus (HBV) 2 months after complete remission.Although he became positive for HB surface antigen at the onset of fulminant hepatitis, he had been negative for HBV serum markers, and only HBV DNA was detected by polymerase chain reaction (PCR) amplification on admission. The original stocks of serum samples from all blood donors were tested again for HBV DNA by PCR, and all samples were negative. This case demonstrates that testing for HBV DNA by PCR is necessary before chemotherapy, because silent HBV carriers are rare and fulminant hepatitis may be induced by chemotherapy in patients with hematologic malignancies.

Budding Process and Fine Structure of Lymphadenopathy-associated Virus (LAV)

The budding process and fine structure of lymphadenopathy‐associated virus (LAV), were studied by indirect immunofluorescence (IF) and electron microscopy (EM). By IF, LAV antigen was seen to be distributed focally within infected CCRF‐CEM cells. Consistent with this finding, electron micrographs showed that LAV particles occurred in a focally aggregated state in a restricted area of the surface of the infected cells. LAV particles possessed bar‐shaped, dense and central or eccentric cores. In addition, two or more cores were occasionally observed in one virus particle, or the cores were sometimes absent when thin sections were examined. The envelope of the virus particles had an irregular structure, although LAV particles were approximately spherical.

Distribution of the Level of Antibody to AIDS-Associated Virus (LAV) in Sera from AIDS or AIDS Related Complex and Japanese Hemophiliacs Infected with AIDS-Associated Virus

The level of antibody to AIDS‐associated virus (LAV) in sera from patients with AIDS or AIDS related diseases (AIDS related complex; ARC) and Japanese hemophiliacs was studied using indirect immunofluorescence. Titer of anti‐LAV antibody in sera from 89 patients with AIDS or ARC ranged between 10 and 40,960 (median: 1,280) and that of 83 Japanese hemophiliacs ranged from 20 to 20,480 (median: 640). The distribution of the level of anti‐LAV antibody in hemophiliacs was similar to that of patients with AIDS or ARC, and no correlation between the titer of antibody and the presence of symptoms of AIDS was observed. Our results suggested that hemophiliacs in Japan might have been infected with live LAV rather than been immunized with inactivated LAV by injection of factor VIII or IX concentrate, and more hemophiliacs in Japan might show symptoms of AIDS in the future as in the United States.

Serum soluble IL-6 receptor levels during the mobilization of stem cells to peripheral blood.

Serum soluble interleukin-6 receptors (sIL-6R) have been demonstrated to play an important role in hematopoiesis. We report here that serum sIL-6R levels reflect proliferative kinetics of the progenitors after stimulation by chemotherapy plus granulocyte colony-stimulating factor. Serum sIL-6R were serially evaluated in 26 courses of peripheral blood (PB) stem cell collections in 16 patients using enzyme-linked immunosorbent assay. Expressions of IL-6R and CD34 on PB mononuclear cells were examined by flow cytometric analysis and expressions of IL-6R mRNA were examined by reverse transcriptase polymerase chain reaction. There were no significant differences between the serum sIL-6R levels on day 0 in patients (27.8 ± 2.1   ng/ml, mean ± SEM) and those in controls (27.5 ± 1.5   ng/ml). Following chemotherapy the serum sIL-6R levels were significantly decreased, reaching a minimal level on day 14 (22.3 ± 1.2   ng/ml, p<0.01) and then significantly increased to above the baseline levels on day 21 (32.0 ± 2.1   ng/ml, p<0.01). Similar oscillations in the number of white blood cells, IL6R + cells, CD34 + cells and colony-forming unit-granulocyte/macrophage (CFU-GM) in PB could be observed and the peak expression of mRNA was compatible with the expression of antigen. Serum sIL-6R levels on day 17 and 19 were positively correlated with the number of CD34 + cells, IL-6R + cells, CFU-GM in PB and the number of collected CD34 + cells in leukapheresis products. In addition, when comparing the 2 groups divided by the number of prior chemotherapies, the status of disease or dose of the mobilizing regimen, the serum sIL-6R levels were significantly increased after day 17 in the group that received fewer courses of prior chemotherapy, the group in complete remission and the group of high-dose chemotherapy. These findings indicated that sIL-6R levels do not reflect the hematopoietic ability in the steady state, or the capability of the hematopoiesis after stimulation. Thus, sIL-6R levels may be a marker for the timing of PBSC collection or the prediction of the number of collected CD34 + cells.

High prevalence of hepatitis C virus genotype la in Japanese hemophiliacs

Abstract Japanese hemophiliacs have received a large number of imported coagulation factors from the USA as well as domestic ones. We investigated the prevalence of hepatitis C virus (HCV) genotypes in 25 hemophiliacs and in 461 patients with chronic hepatitis C in the San-in area, west Japan. The prevalence of HCV genotype la was significantly higher in hemophiliacs (36%, P 〈 0.001) than patients with chronic hepatitis C (0.2%). Furthermore, the mixed infection of different genotypes was significantly more frequent in hemophiliacs (20%, P 〈 0.0001) compared to chronic hepatitis C patients (1%). These results suggest that Japanese hemophiliacs could be infected with HCV genotype la by numerous and repeated infusion of imported coagulation factors from the USA.

Effect of screening of donors for antibodies to hepatitis C virus by second-generation assay on incidence of HCV infection in acute leukemia

Abstract We examined the incidence of hepatitis C virus (HCV) infection in 42 patients with acute leukemia before and after blood donor screening for the HCV Ab using the first-generation assay and the second-generation assay. Although 14 (93%) of the 15 patients before screening and 6 (50%) of the 12 patients after screening by the first-generation assay became seropositive for HCV RNA, none of the 15 patients became seropositive after screening by the second-generation assay. These findings indicate that the second-generation assay is effective in preventing HCV infection in multitransfused patients with acute leukemia.

Effect of Interferon-α on Patients with Previously Untreated Chronic Myelogenous Leukemia in the Early Chronic Phase: Comparison between Interferon-α Continued Patients and Interferon-α Discontinued Patients.

In order to confirm the effect of interferon-α (IFN-α) in inducing a prolonged duration of the chronic phase (CP) on patients with chronic myelogenous leukemia (CML), we retrospectively compared the duration of CP between patients who continued on IFN-α and the patients in whom IFN-α was discontinued before the blast phase. Of the 32 patients not pretreated for CML in the early CP who received IFN-α therapy, 25 continued on IFN-α while seven discontinued the therapy (side effects, 5; resistance, 1; patients refusal, 1). Only four of the 25 patients in whom IFN-α was continued (16.0%) progressed to the blast phase or accelerated phase, but six of the seven patients who discontinued IFN-α (85.7%) progressed to the blast phase or accelerated phase. Fourteen of the 25 patients who continued on IFN-α therapy showed cytogenetic response (complete cytogenetic response, 3; minimal cytogenetic response, 11) whereas 11 patients showed no cytogenetic response. However, non-responders showed a longer duration of CP than the patients whom IFN-α was discontinued. Although elderly patients showed a high incidence of side effects, and some patients progressed early after the beginning of IFN-α therapy, our data clearly demonstrated that in accordance with previous large multi-centric randomized studies the continuation of IFN-α, even in low doses, prevents disease progression.

Replication of Human Immunodeficiency Virus in Two T-Cell Lines and Their Application as Antigens for Immunofluorescence

Two T‐cell lines, TALL‐1 and CCRF‐CEM, were infected with human immunodeficiency virus (HIV), strain LAV, to explore the time course of the appearance of various virus specific antigens, and to establish an antibody assay system by indirect immunofluorescence (IF). These cells were infected with LAV at two different input multiplicity of infection (MOI). Antigens were tested by Western blot analysis (WB) and IF. Antigens for WB were extracted from the infected cells at various times after infection, but pooled sera of American HIV carriers could not recognize gp41 or gp160. Antigen expression was highest in CCRF‐CEM, but, as the antigen for IF, TALL‐1 infected at the MOI of 8.0 was the most suitable 7 days after infection, because it includes a fairly large number of uninfected cells, which served as the internal control.