Due to the advance of chemotheapy and bone marrow transplantation, adult acute leukemia has become a curable disease. In acute myeloid leukemia, the JALSG AML89 study resulted in 77% complete remission (CR) rate in 326 adults and 38% 4.5-year disease-free survival (DFS) in CR cases. However, the result of acute lymphoblastic leukemia in the JALSG ALL87 study is not satisfactory; 84% CR in 116 adults and only 24% 6-year DFS. For acute promyelocytic leukemia (APL), all-trans retinoic acid works remarkably, and the JALSG AML92 study for newly diagnosed APL resulted in 89% CR in 109 adults and 81% 2.5-year DFS.

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When one hundred of c-Kit+Sca-1+Lin- cells obtained from B6. Ly 5.1 mouse were injected into lethally irradiated B6. Ly 5.2, 100% of the mice injected survived. Sequential analyses of recipient's peripheral blood leukocytes revealed presence of Ly 5.1+ myeloid and lymphoid cells even 12 months after the injection. We also examined the fate of these cells in the recipient mouse after transplantation. Time course analyses of the mouse injected with 500 FACS-purified c-Kit+Sca-1+L- cells revealed that spleen is the primary site for expansion of the injected c-Kit+Sca-1+Lin- cells. By day 28 s/p transplantation, more than 50,000 donor type c-Kit+Sca-1+Lin- cells were found in the spleen and over 15,000 cells in the bone marrow. By day 60, number of c-Kit+Sca-1+Lin- cells decreased in the spleen and returned to normal level in the bone marrow. Stem cell activity of the donor derived c-Kit+Sca-1+Lin- cells in the primary recipient was confirmed by in vitro and in vivo colony formation as well as transplantation to the secondary recipient. These results provide evidence that c-Kit+Sca-1+Lin- cells in the bone marrow have capability of self-renew as well as multilineage differentiation potential and that spleen is the primary site of stem cell expansion after bone marrow transplantation in mouse.

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Temporally orderly activation of the cyclin-dependent kinases (cdks) governs the progression and the transitions of the cell cycle in mammalian cells. Macrophages require a specific growth factor, M-CSF/CSF-1, for their proliferation throughout the G1 phase of the cell cycle. Once cells enter S phase, macrophages complete mitosis in the absence of M-CSF/CSF-1. During the G1 phase, cyclin D1 is induced by M-CSF/CSF-1 stimulation and forms enzymatically active complex with cdk 4. The enzymatic activity of the cyclin D1 and cdk 4 complex could be negatively regulated by recently reported inhibitory proteins to determine the timing for entry into S phase in macrophages.

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Inability to distinguish between normal and diseased cells by chemotherapeutic agents causes systemic toxic effects. This problem may be solved by the direct genetic approach using antisense oligodeoxynucleotide (AS ODN) based on the specificity of Watson-Crick base pair formation, if an appropriate disease-specific target can be identified. Since the pioneering works by Zamecnik and Stephenson to inhibit gene expression using AS ODN, recent progress in cloning of pathogenic genes and technical advance in the synthesis of ODN analogues have spurred a research effort dedicated to the development of AS therapy for cancer and viral disease. Chemically-modified ODNs such as phosphorothioate analogues, which are nuclease resistant and considered to be suitable for clinical use, can effectively inhibit the expression of activated oncogenes or the viral replication and lead to the growth suppression of cancer cells and viral genomes in vitro and in experiments using animal models as well. Phase I clinical trials, designed to evaluate the toxicity of these compounds in leukemia or AIDS patients, have already commenced. In these trials, which are now in its infancy, a considerable number of problems will be encountered. However, these hurdles may not be insurmountable.

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Clinical courses and several hematological and serological markers were evaluated in 34 patients with HIV infection (32 with hemophilia and 2 with homosexuality) who had been followed up at Shizuoka Prefecture Children's Hospital for 3 to 13 years, with the reference to the results observed in 1,082 patients with HIV infection registered with the Ministry of Health and Welfare Study Group, Natural History Committee. The results indicated as significant follow-up markers as (1) CD4- and CD8-positive lymphocyte count, (2) IgA, IgG and beta-2 microglobulin, (3) HIVp24 antigen and HIV culture isolation, and (4) serum HIV-RNA. Of these markers, the quantification of serum HIV-RNA seemed particularly useful for prognosis estimation, determination of therapy onset, and efficacy assessment of a given anti-HIV agent. In addition, we encountered 2 HIV-infected patients who seemed to meet the definition of long-term survivors. Factor analysis in these patients will be an important subject of investigation from a clinical point of view as well.

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We heve been isolated HIV strains from blood specimens of HIV infected individuals in Japan for these 6 years. The number of specimens tested reached approximately 1,700 that ninety percent of them were from hemophiliacs repeatedly injected blood products from the United States. More than 300 of field HIV were successfully isolated from the samples. The isolation rates has dicreased to 30 percect in 1993 from 40 percent in 1992, suggestting that treatment with anti-HIV drugs such as AZT and/or ddI may be effective to HIV-infected individuals. Further, both of the viral and genomic sequences of HIV were classified to be clade B virus. The clinical isolates that expressed IHIGPGRAFY sequence at the center of the HIV-V3 domain were found to be neutralized by an anti-clade B-V3 monoclonal antibody, μ5.5. By individual levels, when asymptomatic seropositives have progressed to disease-states, neutralization core motif of GPGR in approximately 6% of the viruses has changed to GPGG and hydrophilic amino acid changed to hydrophobic amino acid, correlating the loss of binding activity to PND-peptide of Japanese Consensus virus. Further, rapid progressors to HIV-induced diseases ahowed decreased activity of the binding antibody.
By using the Japanese consensus sequence of HIV-1, we successfully constructed chimeric protein secretion vectors by selecting an appropriate insertion site of a carrier protein, and established the PND-peptide secretion system in BCG. The recombinant BCG inoculated guinea pigs were inilially screened by delayed-type hypersensitivity (DTH) skin reactions to the PND peptide followed by passive transfer of the DTH by the systemic route. Further, immunization of mice with the rBCG resulted in induction of CTL. The guinea pig immune antisera showed elevated ELISA titer to the PND peptide and neutralized HIVMN, and administration of serum IgG from the vaccinated guinea pigs were effective in completely blocking the HIV infection in Thy/Liv SCID/hu or SCID/PBL mice. In addition, the immune serum IgG neutralized primary field isolates of HIV that match neutralizing sequence motif by peripheral blood mononuclear cell (PBMC)-based virus neutralization assay. The data strongly support that the antigen secreting recombinant BCG system can be used as a tool for development of HIV vaccines.

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Anti-HIV treatment is the major strategy against HIV infection and AIDS. Nucleoside reverse transcriptase inhibitors have been studied extensively, and some of them have been approved for clinical use. Efficacy of non-nucleoside reverse transcriptase inhibitors and HIV-protease inhibitors are also being confirmed. It became clear, however, that all of these agent, allow emergence of drug-resistant HIV mutants when used as monotherapy. Therefore, combination therapy or alternating therapy using these and other new agents may become the main mode of treatment in the future. Clinical trials for HIV vaccines are now being conducted in U.S.A. and other countries. In many of them, immunological responses were confirmed, although clinical benefit was not known yet. Vaccines which induce cellular immunity against broad spectrum of epitopes are desired to overcome viral mutations. Gene therapy is very attractive, and extensive studies are being conducted in many laboratories including ours. Methodology for early and correct diagnosis of opportunistic infections are now developing especially by using molecular technology. Steady improvement in the clinical management of opportunistic infections is achived.

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We investigated the present health care situation in medical institutes of medical doctors who are the members of human immunodeficiency virus type-1 (HIV) -research group headed by Prof. Yamada through mailing-questionaire. It was revealed that there are many HIV-treatment-related problems in medical institutes, included a refusal to accept a newly-diagnosed HIV-infected patients or to conduct surgical treatment on them. Furthermore, it was also clarified that there are quite a many HIV-infected patients who were not informed their HIV-status yet, thus causing secondary-HIV-infection to their sexual partners. From these results, urgent drastic measures to solve these problems are needed for the caring patients living with HIV.

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To establish diagnostic criteria for hypocellular acute leukemia (HL) 32 cases (mean age 67) with 40% or less bone marrow celluarity were analysed and compared with 40 cases of MDS, 27 cases of AML in the elderly (60≥) and 39 cases of AML in the young (60<). The mean bone marrow cellularity was 30% in HL, 85% in MDS, 87% in eldely AML and 95% in young AML, respectively. Thus hypocellularity was evident in HL. Blast % in bone marrow of HL patients was 17-70% in all nucleated cells including lymphocytes (ANC), 36-93% in non-lymphocytic cells (NLC) and 50% or more in all cases in non-erythroid/non-lymphocytic cells (NENLC). Thus maturation arrest of blast cells was evident in HL, which corresponds to that of overt AML. Out of 20 cases treated with low-dose ara-C 13 cases (65%) achieved complete remission, but most of them relapsed early by manifesting hypocellular bone marrow again. In conclusion HL is a distinct clinical subtype of AML in the elderly, which can be clearly defined by 40% or less cellularity and 30% or more blasts in bone marrow.

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Low-grade lymphomas are rare in Japan, comprising about 10% of non-Hodgkin's lymphomas. They are indolent in natural history with median survival of 7∼10 years. Despite a good initial response to radiotherapy or chemotherapy, most patients (Pts), being incurable, ultimately die of their disease. Pts with clinical stage (CS) I/II disease are generally treated with locoreginal radiotherapy. Long-term survival of 60% to 70% and disease-free survival (DFS) of 50% to 55% have been reported. The more precise the staging procedures, the better the results. Chemotherapy used as a primary therapy or as an adjuvant to radiotherapy may improve DFS of pts with CS II with many sites of lymphonode involvement or bulky disease. For CS III/IV disease there is no accepted standard therapy. There is no survival advantage for any of the different first-line treatment methods ranging from initial deferral of treatment albeit in highly selected pts, an alkylating agent ± steroid hormone, to combination chemotherapy (C-MOPP, CHOP, ProMace-MOPP etc) ± irradiation. The international index for prognostication of aggressive lymphoma has also been shown to be applicable to stratification of low-grade lymphomas. In future studies effectiveness of treatment strategies should be investigated with long-term DFS and improved quality of life being set as primary endpoints.

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The problem of diagnosis, prognostic factors and the efficacy of therapies were investigated in 330 patients with multiple myeloma (MM) and 51 patients with benign monoclonal gammopathy (BMG)/monoclonal gammopathy of undetermined significance (MGUS).
Seven out of 51 patients with BMG/MGUS were transformed into MM. The mean time to the transformation was 61.6 months. M protein level in these patients had been gradually and constantly increasing until the transformation in contrast with stable level in non-transformed patients. In MM there was one year difference between median survival from the time of diagnosis and start of chemotherapies. It depended on the deferral of treatment in patients with stage I myeloma.
No difference of survival time was found between initial and diferred therapy for stage I myeloma. Earier therapy is not advantageous in this stage. Stages and immunoglobulin classes of MM were prognostic factors. Stage I or IgG myeloma had the longest survival and stage III or BJP myeloma had the shortest one. The new protocol, DMVM plus natural interferon alpha therapy induced high complete remission rate of 37.1% in initial treatment patients. The survival rate at three years from the treatment was 70%.

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Essential thrombocythemia (ET) is one of the chronic myeloproliferative disorders (MPD), clonal disorders of hematopoietic stem cell. The differential diagnosis from the other MPD and reactive thrombocytosis was described. The treatment of this disease was then stated with emphasis on the importance of control of platelet count. For the control, drugs to suppress the megakaryocytic proliferation are used, including busulfan, hydroxyurea, interferon-α and Anagrelide. The therapy for asymptomatic patients and pregnant women as well as patients with acute hemorrhage and thrombosis was also discussed.

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A 69-year-old woman was admitted to our hospital because of leucocytosis and systemic lymphadenopathy. On admission, white blood cell count was 163,000/μl, most of which consisted of flower-like cells with convoluted nuclei in the peripheral blood. In the abnormal lymphocyte cells surface-marker test by flow cytometry showed that they expressed CD2, CD3, CD4, CD29, CD45RA, and CD38, but not CD8, CD16, and CD25. Serum anti-Human T-lymphotrophic virus type-I (HTLV-I) antibody was negative in particle agglutination, enzyme-linked immunosorbent assay (ELISA) and western-blotting assay. HTLV-I proviral DNA in the abnormal lymphocyte cells was not detected by southern blotting hybridization technique. Moreover, HTLV-I provirus was not detected using a polymerase-chain-reaction (PCR). A monoclonal rearrangement of the TCR-β chain gene was evident by using DNA probe in southern blot hybridization. Because of the rapid progress of the disease, chemotherapy was started immediately after admission. Though, this patient became refractory, and she died about 1 year after admission.

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A 62-year-old man visited our hospital in July 1993, because of a right thigh mass which had grown gradually since two years previously. Physical examination revealed that the mass at the right thigh region, was elastic soft and about 15×10 cm in diameter, without regional lymph node swelling. An ultrasound study showed a hypoechogenic and mesh patterned mass. MRI revealed that the tumor was well defined from subcutaneous adipose tissue and skeletal muscle, indicating that it arose in connective tissue. Angiography demonstrated diffuse hypervascularization of the tumor, and Gallium scintigraphy showed remarkable accumulation at the tumor. Serum IgM was increased, which was proven to be an monoclonal hypergammopathy (IgM, λ). Histological examination of a biopsied specimen obtained from the thigh mass revealed B cell lymphoma, lymphoplasmacytic cell type. The patient achieved a complete remission after surgical treatment following radiation and combination chemotherapy.

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A 20-year-old female was admitted because of high fever, hepatosplenomegaly, severe hepatic dysfunction and coagulopathy. Peripheral blood showed pancytopenia and granular lymphocytes bearing the natural killer cell phenotype (CD2+CD3-CD16+CD56+CD57-TCRαβ-TCRγδ-) constituted 97% of leucocytes. Southern blot analysis of DNA obtained from peripheral blood mononuclear cells showed germ-line configuration of TCRβ, γ and δ chain genes. EBV-DNA was detected in a single episomal form by using EBV-terminal repeat probe. Bone marrow findings were consistent with hemophagocytic syndrome and administration of VP-16 was effective transiently. After ten months she died from massive gastrointestinal bleeding. An in situ hybridization study identified EBV-RNA (EBER-1) in atypical lymphocytes infiltrating bone marrow, spleen and lymph nodes. Sections of liver showed steatosis and infiltration of Tcells (CD3+and EBER-1-negative) in the portal areas and few atypical lymphocytes in sinusoids. The patient developed an EBV-associated clonal proliferation of natural killer (NK) cells, but the clinical features were suggestive of chronic active EBV infection or virus-associated hemophagocytic syndrome (VAHS) rather than leukemia. Bone marrow transplantation for NK cell leukemia is an issue to be discussed.

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