We analyzed immunoglobulin (Ig) gene rearrangements in 69 patients with B-cell malignancies by Southern hybridization. We used 5 probes which covered J_H 5'Sμ and Sγ3 loci of the Ig heavy (IgH) chain gene, and Cκ and κde loci of the Ig light (IgL) chain κ gene, respectively. DNA rearrangements were observed in 68 out of the 69 patients using the J_H probe. In 97% (33/34) of patients with non-Hodgkin's lymphoma (NHL), 100% (5/5) of chronic lymphocytic leukemia (CLL), 42% (10/24) of non-T cell acute lymphoblastic leukemia (non-T ALL), and in 0% (0/4) of chronic myelogenous leukemia lymphoid crisis (CML-LBC), the rearrangements were detected by Cκ and/or κ de probes. Furthermore, the genotype of the light chain was defined by rearrangement patterns of these 2 probes.
The by using the 5'Sμ-probe, class switch recombinations were detected in 76% (25/33) of NHL, 20% (1/5) of CLL, 17% (4/24) of non-T ALL, and in 0% (0/4) of CML-LBC. Among them, 1 case of IgM NHL and 3 cases of double bearer NHL had rearranged on both IgH alleles by the constant region. The results of this study indicate that genotypes correspond well to phenotypes of B-cell malignancies and that the determination of genotype will be useful for making an exact diagnosis of B-cell malignancies.

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Preventive effects of hemorrhagic cystitis by the use of sodium 2-mercaptoethane sulfonate (MESNA) was evaluated in 40 children undergoing peripheral blood stem cell autografts (PBSCT) after marrow-ablative chemotherapy which included high-dose cyclophosphamide (CY, 50mg/kg×2). Fifteen patients received MESNA (group A) and 25 did not (group B), and all received concomitant hyperhydration (3,000ml/m²/day). No renal dysfunction or toxicity attributed to the use of MESNA was observed in either group of patients. Transient hemorrhagic cystitis developed in one of the 15 group A (6.7%) and 3 of 25 (12.0%) group B patients but there was no statistical significance. Although the results may suggest that MESNA is a contraversial agent in preventing hemorrhagic cystitis caused by CY when hyperhydration protocol is used, further observation with a larger number of patients is required to establish a firm conclusion.

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The expression of cytoplasmic antigens in 77 cases of acute leukemia were analyzed by flow cytometry using the following monoclonal antibodies: CD3, CD22, anti-myeloperoxidase (MPO-7) and anti-μ-heavy chain.
CD22 antigen was detected in the cytoplasm of all non-T-ALL patients excluding one not-tested patient. In two patients with unclassified ALL, surface CD22 antigen was not expressed but cytoplasmic CD22 antigen was strongly expressed.
Three out of 9 patients with common ALL were cytoplasmic μ-heavy chain-positive, so these patients were diagnosed as Pre-B ALL. In four out of 8 patioents with T-ALL, CD3 antigen was not expressed on the cell surface membrane. Howover all of T-ALL patients excluding one non-tested patient were cytoplasmic CD3-positive.
The cytoplasmic expression of myeloperoxidase antigen was detected in twenty out of 21 patients with acute non-lymphoblastic leukemia (ANLL). One megakaryocytic leukemia patient was MPO-negative. In two ANLL patients, the percentage of MPO for conventional cytochemical staining was undetectable or low, but MPO antigens were positive (77% and 70%) for flow cytometric analysis. All of 46 non-T ALL patients were cytoplasmic MPO-negative, however 4 out of 10 T-ALL patiens were cytoplasmic MPO-positive.
The study proved that the analysis of cytoplasmic CD3, CD22, μ-chain and MPO antigens were very useful to define the cell lineage of leukemia and to classify ALL and ANLL. It is nessesary to study further whether the expression of MPO in the cytoplasm of T-ALL was non-specific reaction or whether MPO precursors are expressed in the cytoplasm of T-ALL.

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This study was designed to investigate the pathogenic relevance of platelet-associated autoantibodies in chronic ITP, since the titer of platelet-associated autoantibodies does not appear to correlate with the severity of the disease. Employing a direct immunoprecipitation procedure, we examined platelets from three ITP patients with platelet-associated autoantibodies against GPIIb-IIIa and an unidentified 56kD protein before and after splenectomy. In two patients, platelet-associated autoantibodies disappeared after splenectomy, and these two patients attained complete remission. In one patient, however, the amount of platelet-associated autoantibodies did not decrease after splenectomy. Although this patient's platelet count transiently increased to 500×10³/μl after splenectomy, it decreased to 55×10³/μl within a short time. These findings suggest that platelet-associated autoantibodies play a key role in platelet destruction in chronic ITP.

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We described two cases having erythroid hypoplasia and pancytopenia, respectively, caused by human parvovirus B19 (PVB19) infection on chemotherapy. The first patient was a seven-year-old boy with Non-Hodgkin's lymphoma. He has obtainod complete remission with LSA₂L₂ protocol, but, immediately after this remission, he suddenly developed high fever, erythema on cheeks and severe anemia without reticulocytes. We concluded that the cause of anemia and other symptoms were due to PVB19 infection because PVB19 DNA in his serum was detected by dot blot hybridization and polymerase chain reaction, although specific antibodies to PVB19 remained absent. He received gamma-globulin intravenously (200mg/kg/day for 5 days). The fever and erythema were improved promptly, but viremia and anemia lasted a few weeks and the specific antibodies to PVB19 were negative for about two months thereafter. The second patient was a nine-year-old boy with common ALL in remission. Pancytopenia suddenly occurred under maintenance therapy. Because IgM to PVB19 was detected in his serum, the patient was diagnosed to have temporary pancytopenia due to PVB19 infection. In conclusion, it is important to confirm specific antibodies and PVB19 DNA in serum when anemia or pancytopenia of unknown cause occurs in immunocompromised patients receiving chemotherapy.

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A 33-year-old woman with AML (M4) resistant to chemotherapy received syngeneic marrow graft from her identical twin following high dose busulfan and etoposide. However, the relapse was confirmed on the 60th day after the procedure. Since she failed to achieve remission despite intensive chemotherapy, a second BMT from the same donor was performed following total body irradiation and high dose etoposide on the 126th day after the initial BMT. At this time, cyclosporine (1 mg/kg/day) was administered to induce graft-versus-host disease (GVHD). Skin rash appeared on the 18th day after the 2nd BMT, and biopsy from the rash on the 23rd day showed a typical picture of cutaneous GVHD (grade 2) and there was no evidence of viral infection. On the 36th day after the 2nd BMT, the patient died of veno-occlusive disease. Although graft-versus-leukemia effect in this patient could not be evaluated because of early death, the induction of GVHD with cyclosporine might be effective to reduce the relapse rate after syngeneic or autologous BMT. Further studies are required to confirm this effect.

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A 47 year-old man with selective IgA deficiency (SIgAD) consulted us in November 1981, with complaints of leg edema and common cold-like symptoms and was diagnosed as SIgAD based on data of his serum protein (IgG 2,160 mg/dl, IgM 65 mg/dl, no detectable IgA). Later in July 1989, he was admitted with edema and ascites. Laboratory examinations showed; total protein 4.6 g/dl, albumin 1.26 g/dl, IgG 2,375 mg/dl, IgM 38 mg/dl, no detectable IgA, C3 22 mg/dl, C4 6 mg/dl, antinuclear antibody 80X, anti dsDNA antibody 4.5 U/ml, anti IgA antibody 258%, and lymphocytopenia. Co-culture of lymphocytes from the patient and normal subject revealed deficiency of IgA synthesis in his B cell populations. Systemic lupus erythematosus was suggested based on the findings of skin biopsy, renal damage, oral ulcer, decreased complements, autoantibody and lymphocytopenia. We could not give him conventional products of albumin and frozen plasma because he had anti IgA antibody. Instead, we administered concentrated autogenous ascitic fluid and prednisolone. His ascitic fluid disappeared and complements and albumin in his serum normalized. He has continved in good condition and is beeing treated as an outpatient.

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We report a case of intermediate lymphocytic lymphoma (ILL) with multiple lymphomatous polyposis. A 56-year-old man presented with general fatigue and bloody stool. Physical examination showed cervical and axillary lymphadenopathy, bilateral tonsillar hypertrophy, and moderate splenomegaly. Leukocyte count was 9,570/μl with 11% abnormal cells, infiltration of which was observed in the bone marrow too. Examinations of the gastrointestinal tract revealed diffuse small polypoid lesions throughout the stomach and the entire large bowel. The biopsied specimens from both the stomach and large bowel showed diffuse infiltration of medium-sized lymphoid cells in the submucosa and the lamina propria. Lymph node biopsies showed ILL (mantle zone lymphoma). The phenotype of lymphoma cells was CD5 (+) CD10 (-) CD19 (+) CD20 (+) CD21 (+), and sIgμ δ-λ. The patient was initially given the multiple agent chemotherapy, which did not improve the peripheral blood findings and was switched to the regimen that comprised of etoposide and prednisolone. The patient's lymphoma is well controlled by this regimen 35 months after diagnosis.

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Two cases of acute leukemia with a t(6;9)(p23;q34) chromosome abnormality are reported. The first case was a 34-year-old female who was hospitalized in October 1989. A diagnosis of FAB-M1 was made. Chromosomal analysis of the bone marrow cells showed a 46, XX, t(6;9)(p23;q34). Complete remission was achieved after two courses of BHAC-DMP therapy. In September 1991, at the time of relapse, chromosomal analysis revealed two abnormal clones consisting of a 46, XX, t(6;9)(p23;q34), -12, -17, +der (12) t(12;17)(p11.2;q11.2) with a residual normal clone. She died in February 1992. The second case was a 42-year-old male who was hospitalized in January 1990. He was diagnosed as having RAEB. Chromosomal analysis of the bone marrow cells showed 46, XY, t(6;9)(p23;q34). Three months later, the disease progressed to acute leukemia accompanied by leg ulceration with leukemic cell infiltration. Small-dose ara-C therapy was given, but with no effect. After two subsequent courses of therapy with low-dose etoposide, complete remission was achieved. Four months later, relapse occurred, and the patient died of sepsis in February 1991. In the literature, 31 cases of myeloproliferative disorders with t(6;9) have been reported.

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This report describes 4 cases of T-cell-associated CD7-positive acute myeloid leukemia (AML). Myelo-peroxidase staining of blasts was negative in 2 cases but became positive during their courses. In all cases, the myeloid determinants CD13 and/or CD33 were associated with CD7 expression. Other B-lymphoid (CD10, CD19) or T-lymphoid (CD2) markers were negative. In three cases, dual fluorescence analyses showed co-expression of CD7 and CD13 (CD33). Clinically, compared with CD7^- AML, these CD7⁺ AML patients presented higher leukocyte and blast counts in peripheral blood. All patients achieved complete remission with chemotherapeutic regimens for AML, but 3 relapsed within a short time. Systemic lymphadenopathy was found in 2 cases, and interestingly, the surface markers of the lymph-node in one case were CD7⁺CD33^-. These cases of CD7⁺ AML may represent a distinct subgroup that arises from particular, less different myeloid precursors, and may have poor prognosis.

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A 25-year-old man with acute lymphocytic leukemia (ALL) had received bone marrow transplantation (BMT). Testicular relapse occured 6 months after BMT as the first relapse, and a couple of weeks later, bone marrow relapse developed. At that time, karyotypic analysis showed Ph-chromosome which was not observed before the first remission and DNA analysis revealed rearrangements of minor BCR, IgL, IgH, TCR and TCR genes. Testicular relapse often develops in ALL of children after BMT. However, some reports, including this report, indicate that testicular relapse following BMT can also occur in adults, so that the incidence of testicular relapse could be reduced either by total body irradiation (TBI) or testicular irradiation before transplantation. Incidence of Ph-positive chromosome following BMT is a key to illuminate the mechanism of the Philadelphia chromosome. Rearrangements of Ig and TCR gene can be considered as an abnormal gene rearrangement occuring at an early stage of B cell proliferation.

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A rare case of dermatomyositis and thrombotic thrombocytopenic pupura (TTP), which responded dramatically to high-dose gamma globulin and vincristine is presented. A 42-year old man was admitted for evaluation of polymyalgia and skin change of the face and fingers. Findings of muscle biopsy was consistent with the diagnosis of dermatomyositis. During the course of his hospital stay, he had diffuse purpura, hematuria, high fever, and his consciousness became disturbed. The hemoglobin level and the platelet count decreased. Based on microangiopathic hemolytie anemia, thrombocytopenia and neurological abnormality, a clinical diagnosis of TTP was made. Therapy included high-dose gamma globulin, vincristine, corticosteroids and dextran. One week later, his consciousness became clear, hematological findings improved, and prolonged remission has been maintained for more than 19 months at the time of this report. This case suggests that gamma globulin and vincristine are effective in some with TTP cases.
Case reports that have appeared in the Japanese literature are summarized and reviewed in terms of treatment.

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An adult T-cell leukemia (ATL) accompanied with eosinophilia is described. A 75-year-old female was admitted to our hospital because of lymphadenopathy. Her leukocyte count was 73,300/μl, with 35.5% abnormal lymphocytes and 19% eosinophils. A majority of lymphocytes expressed CD4+CD8-. Acute ATL was diagnosed, since anti-HTLV-I antibody in her serum and monoclonal integration of HTLV-I proviral DNA in her peripheral mononuclear cells were detected. She was treated with THP-adriamycin, cyclophosphamide (CPA), and vincristine (VCR). Abnormal lymphocyte and eosinophil counts decreased and there was improvement in the lymphadenopathy. However she then complained of lymphadenopathy again. Her leukocyte count rose to 76,300/μl, with 89% abnormal lymphocytes. Combination therapy with CPA, VCR, and doxorubicin was started and there was a temporal regression in lymphadenopathy, but her lymphadenopathy recurred and she died. The activity of granulocyte-macrophage colony-stimulating factor (GM-CSF) was detected in the supernatant of the cultured ATL cells, although interleukin-3, interleukin-5, and GM-CSF activities were not detected in her serum. It seems likely that the secretion of GM-CSF by ATL cells are responsible for the eosinophilia.

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A 64-year-old man had been found to have primary splenic lymphoma (stage III) seven years after the diagnosis of rheumatoid arthritis (RA). Histological diagnosis of the lymphoma was diffuse, medium sized cell type (LSG) or intermediate lymphocytic lymphoma (ILL). Splenectomy and ten courses of CHOP regimen produced continuing remission. After tow years, he suffered from peripheral neuropathy due to vasculitis of polyarteritis nodosa (PN) type. He was treated with prednisolone (PSL) and cyclophosphamide (CPM) for malignant rheumatoid arthritis. One year later, evaluation for intermittent hematuria revealed bladder cancer and he underwent total cystectomy. He has been treated with small doses of PSL under observation. The high incidence of ILL in lymphomas developing in patients with autoimmune diseases of the thyroid and salivary glands has been reported. This case suggests an association between antecedent RA and splenic lymphoma, the influence of splenectomy and chemotherapy on occurrence of rheumatoid vasculitis, and a causal relationship between CPM and bladder cancer.

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A 79-year-old man who had been diagnosed as having sarcoidosis when he was 63 year old, was admitted to our hospital because of marked thrombocytosis and leukocytosis in July 1991. The low neutrophil alkaline phosphatase (NAP) score, presence of Philadelphia (Ph¹) chromosome in the bone marrow cells, and M-BCR rearrangement by Southern blot hybridization were observed. He was diagnosed as having chronic myelogenous leukemia complicated with sarcoidosis. The coexistence of sarcoidosis and leukemia has rarely been reported. It is difficult to discuss that there is not causal association between of them.

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