Natural killer cell activity and cytokine production as prognostic factors in adult acute leukemia.

We studied the natural killer (NK) cell activity and in vitro production of the cytokines which can enhance NK activity (interleukin-1 beta (IL-1 beta), interferon gamma (IFN gamma), and interleukin-2 (IL-2)) after stimulation in 44 patients with acute leukemia (AL) and 14 normal controls. We also studied the influence of these parameters on relapse and the relapse-free survival (RFS) (after the date of assay) of the AL patients. The NK activity and the production of cytokines in the peripheral blood mononuclear cells (PBMNC) from 16 patients at the untreated or relapsed stage as well as from 12 patients after consolidation were significantly lower than those from controls (both P<0.01), and those from the 16 patients at maintenance or off treatment were also significantly lower than those from the controls (P<0.01 or P<0.05). RFS after the date of assay of the patients in remission with NK activity above the median value was significantly longer than that of the patients below the median (P<0.05). The production of cytokines in the PBMNC from patients who showed continuous complete remission for at least 6 months was higher than that from the patients who relapsed early. These findings suggest that impaired NK cell function and cytokine production are associated with early relapse of AL regardless of remission status.

Clinical significance of serum soluble interleukin-2 receptor in chronic myeloproliferative disorders.

Serum soluble interleukin-2 receptor (sIL-2R) levels were determined in patients with chronic myeloproliferative disorders (CMPD): 18 with chronic myelogenous leukemia in chronic phase (CML in CP), seven with CML in accelerated phase (AP) or blastic crisis (BC), six with polycythemia vera (PV), eight with essential thrombocythemia (ET), one with primary myelofibrosis (PMF), and 50 controls. The mean (+/-S.E.M.) levels were higher in CMPD than in controls (CML in AP or BC, 2693 +/- 694 U/ml, P < 0.0001; CML in CP, 792 +/- 63 U/ml, P < 0.0001; PV 553 +/- 89 U/ml, P < 0.05; ET, 449 +/- 56 U/ml; PMF, 628 U/ml vs. controls, 395 +/- 25 U/ml). Patients with CML in CP had significantly higher serum sIL-2R levels than patients with ET (P < 0.005), and levels were markedly elevated in AP and BC (P < 0.001). Serum sIL-2R levels were positively correlated with WBC count and lactic dehydrogenase in CMPD, and in CML in CP. Serum sIL-2R levels in CMPD were negatively correlated with RBC and platelet counts. Serum sIL-2R levels were significantly lower in patients with CML in CP who showed a cytogenetic response after interferon (IFN) therapy than in those who showed no response (P < 0.05). These findings suggest that a high serum sIL-2R level reflects the leukocyte growth in CMPD and is useful both for differentiating CML from other CMPD and for predicting the response to IFN therapy in CML.

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.

Serum-soluble c-kit levels during mobilization of peripheral blood stem cells correlate with stem cell yield.

c-Kit is expressed in hematopoietic stem cells and plays an important role in hematopoiesis. In 16 patients with malignancies, serum-soluble c-Kit levels and the expressions of c-KIT messenger RNA (mRNA) and protein in peripheral blood mononuclear cells were analyzed serially during 26 courses of peripheral blood stem cell (PBSC) mobilization after granulocyte colony-stimulating factor administration following chemotherapy for PBSC harvest. Serum-soluble c-Kit levels were significantly lower in patients than in controls (179.7+/-17.7 arbitrary units [AU]/mL versus 274.5+/-18.9 AU/mL; P < .001), decreasing after chemotherapy (167.7+/-18.2 AU/mL), increasing from day 14, and peaking at day 19 (193.3+/-16.4 AU/mL). The numbers of both c-Kit+ cells and CD34+ cells and granulocyte-macrophage colony-forming units in peripheral blood peaked at day 17, following the peak of the expression of c-KIT mRNA. Serum-soluble c-Kit levels showed a significant positive correlation with the numbers of CD34+ cells in both peripheral blood and leukapheresis products (r = 0.553, P < .01, and r = 0.640, P < .001, respectively) and changed at higher levels in patients with large numbers of PBSCs versus patients with small numbers of PBSCs (P < .05). Serum-soluble c-Kit may reflect the capacity for hematopoiesis after chemotherapy and may be useful in predicting the number of PBSCs that can be mobilized and harvested after mobilization, as well as for monitoring the timing for PBSC harvest.

An improved method for the detection of HIV antigen in the blood of carriers.

The measurement of HIV antigen levels in sera or plasma of HIV-infected individuals is critical for determining the existence of antigen or infectious virus before seroconversion and for prognosis. Pretreatment of sera or plasma of HIV carriers by heating at 70 degrees C for 10 min at an acidic pH enabled us to estimate antigens efficiently in immune complexes. This procedure will also be useful in determining antigen levels in HIV carriers more precisely.

[Improvement of thrombocytopenia by administration of M-CSF in a patient with relapsed Philadelphia chromosome-positive acute lymphoblastic leukemia].

A 55-year old male, diagnosed as Ph1 (+) ALL in July 1992, subsequently achieved complete remission. However, he was admitted again in November 1993 because of relapse. Although blasts in bone marrow decreased following chemotherapy and G-CSF administration, thromocytopenia remained, and he needed platelet transfusions every other day. After he began receiving M-CSF, the platelet count increased and platelet transfusions became unnecessary. M-CSF may be useful for thrombocytopenia after chemotherapy in Ph1 (+) ALL.

[Nonproducing myeloma without evident bone lesion].

A 76-year-old female was admitted to our hospital because of anemia. Complete blood count was as follows: RBC 2.37 X 10(6)/microliters, Hb 7.7 g/dl, WBC 2,600/microliters, Plt 105 X 10(3)/microliters. A bone marrow aspirate revealed 40.8% plasmacytoid cells showing the characteristics of plasma cells by electron microscopy. Total serum protein was 5.4 g/dl. Monoclonal protein was not observed by electrophoresis. On immunoelectrophoresis, M-bow was not observed in the serum or in 50-fold concentrated urine. The plasma cells were negative for cytoplasmic IgG, M, A, E, D, kappa or lambda by immunoperoxidase studies. Although radiologic studies of the bones did not reveal destructive or punched out lesions, we diagnosed this case as a nonproducing myeloma and the patient responded to MP therapy. This case was considered interesting as regards the pathological entity of myeloma.

[Secondary porphyrinuria].

The condition that the porphyrins are excreted into the urine due to impairment of bile excretion, is called as secondary porphyrinuria. The main porphyrin is coproporphyrin, there fore the condition is called as secondary coproporphyrinuria. Secondary porphyrinuria is associated with various disorders, such as hepatobiliary diseases, hereditary hyperbilirubinemia, intoxications, blood and metabolic diseases, but the etiology of secondary porphyrinuria is unclear. Coproporphyrin is divided into two isomers, one is coproporphyrin-I type and the other is coproporphyrin-III type. In normal human urine, coproporphyrin-III type is predominant, and the ratio of coproporphyrin-I to total coproporphyrin is 10-50% (% as isomer I), while in urines of hepatobiliary diseases, the ratio is 40-60%. In Dubin-Johnson syndrome, the ratio is 80-100%. Coproporphyrinuria in hepatocellular carcinoma and alcoholic liver disease may be different from that in hepatobiliary diseases, often associated with uroporphyrinuria as well as coproporphyrinuria. Coproporphyrinuria arising in blood and metabolic diseases must be taken into account of associated liver diseases.

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.

Prevalence and transmission of human immunodeficiency virus in Japan.

The routes of transmission of human immunodeficiency virus (HIV) can be divided into two categories, 1. sexual transmission (male-to-male, male-to-female and female-to-male) and 2. parenteral transmission by blood or blood products. Among 488 Japanese hemophiliacs, 165 (33.8%) were seropositive and were infected by the injection of factor VIII or factor IX manufactured from American sources. The rates of infection among hemophiliacs were 21.8% (19/87) and 36.8% (43/117), respectively for 1984 and 1985. Sera of 10,272 volunteer blood donors were all negative for anti-HIV. There have been 16 confirmed AIDS cases in Japan consisting of 8 male homosexuals and 8 hemophiliacs, so it can be concluded HIV infections have exclusively occurred among male homosexuals and hemophiliacs. But there are chances to transmit HIV by blood transfusion if the screening of whole blood units is not implemented in the near future.

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.

Antibodies to human immuno-deficiency virus and human T-cell leukemia virus type I in Japanese patients with hematologic malignancies.

One thousand six hundred and seventy-four blood samples drawn between January 1980 and April 1986 from 1454 Japanese, including 251 leukemia, 409 lymphoma, 76 adult T-cell leukemia and 25 benign lymphadenitis patients, were tested for antibodies to HIV and HTLV-I. No patient with lymphadenitis or lymphoma associated with HIV infection was found. In 87 patients with acute and chronic leukemias who had received multiple transfusions, 8 were positive for anti-HTLV-I antibody after blood transfusions amounting to 115 units, on average, while no patient was positive for anti-HIV antibody. Overall, no sample was positive for anti-HIV antibody, whereas 153 (10.5%) were positive for anti-HTLV-I antibody. These results indicate that the transmission of HIV by blood transfusions is far less prevalent than that of HTLV-I in Tokyo at present.

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.

Liver dysfunction in patients with acute myelogenous leukemia: studies on patients not infected with hepatitis C virus during intense therapy.

Liver dysfunction often occurs during chemotherapy for AML, but the etiologies are many and varied. To determine liver dysfunction that is not related to HCV, liver function during intense therapy for one week after complete remission was studied in eight patients not infected with HCV (38 courses) and six HCV-infected patients (19 courses) with AML. There were remarkable differences in changes of ALT levels among HCV-infected patients. ALT level changes among patients not infected with HCV were similar. Changes in mean serum ALT levels in HCV-infected patients occurred at higher serum levels as compared with those in patients not infected with HCV. The mean serum ALT levels in patients not infected with HCV significantly increased at one week (45 +/- 5 IU/l) and further increased at two (58 +/- 8 IU/l) and three weeks (57 +/- 5 IU/l) as compared with pretreatment levels (24 +/- 21 IU/l) (p < 0.001, p < 0.001, p < 0.0001, respectively). ALT levels returned to normal at four weeks. During 31 of 38 courses (81.6%) in patients not infected with HCV, febrile episodes occurred at three weeks. The mean serum ALT levels in patients with febrile episodes were significantly higher than those in patients without febrile episodes at three weeks, and serum ALT levels at three weeks showed a significant positive correlation with CRP levels at three weeks. These findings indicate that liver dysfunction during chemotherapy for AML is due to hepatocellular injury, and infection or inflammatory cytokine induced by infection results in the worsening of the liver dysfunction.

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

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.

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; patient's 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.

Expression and phosphorylation of BiP/GRP78, a molecular chaperone in the endoplasmic reticulum, during the differentiation of a mouse myeloblastic cell line.

To determine the functional significance of endoplasmic reticulum chaperones in hematopoietic cells, we analyzed the expression and post-translational modification of BiP/GRP78 and GRP94 as well as the cytoplasmic chaperones HSP70 and HSC70 during the differentiation of a mouse myeloid leukemia cell line, M1. The amounts of BiP/GRP78 and GRP94 increased several-fold when M1 cells were induced to differentiate into macrophage-like cells by treatment with interleukin-6 (IL-6). Synthesis began to increase at 4 hr after IL-6 treatment. The phosphorylated form of BiP/GRP78 increased during the later stages of differentiation. These data suggested that the chaperone activity of BiP/GRP78 and GRP94 may be needed for differentiated macrophage-like cells or for the differentiation event itself, and that functionally different BiP/GRP78 accumulate during the differentiation of M1 cells.