Toshiharu Tamaki

Demonstration of high-affinity interleukin-2 receptors on B-chronic lymphocytic leukemia cells: functional and structural characterization.

SummaryFunctional and structural characteristics of interleukin 2 (IL-2) receptors on B-cell chronic lymphocytic leukemia (B-CLL) cells were analyzed by a proliferation assay, IL-2 binding assay and cross-linking study. In the3H-thymidine incorporation assay, purified B-CLL cells from four out of sixteen cases, in which the percentage of Tac antigen (Tac Ag) positive cells in peripheral blood lymphocytes ranged from 0 to 48.8%, responded to IL-2 (100 U/ml) after both 3- and 6-day incubation. No relationship was found between the responsiveness to IL-2 and the percentage of Tac Ag positive cells. In the radiolabeled IL-2 binding assay, however, B-CLL cells from all seven cases examined, including three cases with mitogenic response to IL-2 and four cases without mitogenic response, were shown to have both high- and low-affinity receptors. The number of high- and low-affinity receptors per cell ranged from 29–186 and from 420 to 1,800, respectively. Furthermore, with the affinity cross-linking method p55 (Tac Ag) and p70/75 were found even in cases without mitogenic response in their B-CLL cells. In conclusion, the B-CLL cells so far examined possessed high-affinity IL-2 receptors consisting of p55 and p70/75; nevertheless, this was not sufficient to respond to the mitogenic signal of IL-2.

Surface expression of human myeloma cells: an analysis using a panel of monoclonal antibodies

Neoplastic cells of 18 patients with multiple myeloma were studied using a panel of 6 monoclonal antibodies to B cells and monospecific antisera against the light chain types of immunoglobulin. OKT10 bound to the myeloma cells of all the patients, although only a small percentage of the cells reacted in 3 instances. Monoclonal sIg was present in 5 patients. HLA-DR antigen detected with OKIa-1 was found in 5 patients. B1 bound to a small percentage of the myeloma cells only in 2 patients who had received treatment. The B1+ cells were always sIg+. In 3 patients, BA-2 reacted with the myeloma cells. BA-1 and BA-3 invariably did not react with the cells. Myeloma cells with B cell markers were found more frequently in treated patients than in untreated patients. The RPMI8226 line was also studied and found to react with OKT10 and BA-2. The results of this study show the presence of phenotypic variety in myeloma cells.

Successful treatment of refractory subcutaneous panniculitis-like T-cell lymphoma with allogeneic peripheral blood stem cell transplantation from HLA-mismatched sibling donor.

Subcutaneous panniculitis-like T-cell lymphoma (SPTCL) is an uncommon type of peripheral T-cell lymphoma (PTCL) usually presenting with erythematous, subcutaneous nodules and fever [1]. It is considered as a cytotoxic T-cell neoplasm with cyotoxic granules and can be cytochemically identified using granzyme B, perforin or TIA-1 [2]. Some cases of SPTCL might be indolent for months or years, but SPTL frequently has an aggressive course, and is often complicated by fatal hemophagocytic syndrome (HPS) [3]. Among the cases of SPTCL, resistance to anthracyclin-based therapy generally means a poor prognosis [4]. The use of high-dose therapy (HDT) with autologous stem cell transplantation (SCT) in patients with SPTCL who are refractory to anthracyclins has yielded good results [4]. However, a cure can only be expected if the disease is at least sensitive to HDT [5]. Allogeneic SCT, however, might yield a cure even in chemo-refractory lymphoma patients via a graft-vs.-lymphoma (GVL) effect [6]. We report a case of successful use of an allogeneic peripheral blood stem cell transplantation (allo-PBSCT) from an HLA-mismatched sibling donor in a patient with SPTCL who was refractory to conventional chemotherapy. In August 2000, a 17-year-old woman presented to our hospital because of an intermittent fever and a painful swelling of her hip and right thigh. A biopsy specimen of cutaneous tissue from her right hip was diagnosed as cytophagic histiocytic panniculitis. She was treated with oral prednisolone, which resolved the skin lesions. Three months later, she once again presented with persistent high fever and multiple subcutaneous indurations. Oral prednisolone and etoposide were administered, but only a temporary effect on the skin lesions and fever was seen. A second incisional biopsy of the subcutaneous tissue of her right thigh was performed on 19 March 2001. The biopsy specimen showed massive neoplastic lymphoid infiltration of the subcutaneous fat compatible with a diagnosis of SPTCL (Figure 1A). Immunohistochemically, the neoplastic cells were positive for CD3 and CD45RO (UCHL-1), positive for granzyme B, partially positive for CD4 and CD56 and negative for CD20. A test for Epstein – Barr virus using EBER-1 in situ hybridization was negative. A full blood count revealed mild pancytopenia (white blood cell of 2.4610/l, hemoglobin of 111 g/l and platelets of 74610/l). Her serum lactate dehydrogenase level was 1304 IU/l (normal range 150 – 460 IU/l). Bone marrow aspiration revealed marked hemophagocytosis by activated macrophages without involvement of lymphoma cells (Figure 1B). Two cycles of CHOP-E (cyclophosphamide, adriamycin, vincristine, prednisolone and etoposide) resolved the skin lesions, fever, and HPS. However, 2 weeks after the second course of CHOP-E chemotherapy, the skin lesion and fever recurred. Several treatments, such as high-dose etoposide (500 mg/m on days 1 – 3), L-asparaginase (6000 U/m on days 1 – 7), L-asparaginase (6000 U/m on day 2) plus cytarabine (Ara-C; 3 g/m every 12 h on days 1 – 2), and high-dose Ara-C (1.5 g/m every 12 h on days

A case of T‐cell chronic lymphocytic leukemia (T‐CLL) expressing a peculiar phenotype (E+, OKM1+, leu 1+, OKT3− and IgG EA−)

A case of chronic leukemia is reported. Malignant cells had the morphology of lymphocytes and an affinity for skin. Cytochemically, they were peroxidase negative, and NaF‐resistant α‐naphthyl esterase positive. The peripheral mononuclear cells formed E‐rosettes and reacted with Leu 1 and OKM1. They were unreactive with OKT3, OKT4, OKT6, OKT8, and OKIa1. The cells did not possess surface IG, C3b receptors, or IgG Fc receptors (EAIgG). They responded weakly to phytohemagglutinin, and did not respond to concanavalin A. The cells did not adhere to Petri dishes or phagocytose latex beads. It is concluded that it was a case of OKT3−, OKM1+, EAIgG− T‐cell chronic lymphocytic leukemia. This suggests that OKT3 does not recognize all peripheral T‐cells, and that OKM1 is not specific for the monocyte—myeloid lineage. These cells may be useful in the characterization of E+, OKM1+ subset of peripheral mononuclear cells.

Detection of 14q32.33 translocation and t(11;14) in interphase nuclei of chronic B-cell leukemia/lymphomas by in situ hybridization.

Abnormalities of chromosome 14 involving band q32.33 are among the most commonly observed cytogenetic alterations in B‐cell malignancies. To assess the incidence and pathogenetic implications of 14q32.33 translocation in chronic B‐cell leukemia/lymphomas, we performed fluorescence in situ hybridization (FISH) analysis with variable region (VH) and gamma constant region (Cγ) gene probes in 37 patients with these disorders. Chromosome 14q32.33 translocation was detected in 2 of 18 patients with chronic lymphocytic leukemia (CLL), 1 of 2 with CLL of mixed cell types (CLL/PL), 1 of 2 with pro‐lymphocytic leukemia (PLL), 5 of 6 with leukemic mantle‐cell lymphoma (MCL), 2 of 7 with splenic B‐cell leukemia/lymphoma of possible marginal zone origin (SBLL) and 2 with leukemic follicular lymphoma (FL). To further characterize 14q32.33 translocations in these patients, we developed a new procedure using double‐color FISH with PRAD1, BCL2, VH and Cγ gene probes. Chromosome t(11;14) was detected in 1 patient with CLL/PL, 1 with PLL and 5 with MCL. Chromosome t(14;18) was detected in 2 patients with FL. In a PLL patient with t(11;14), the cosmid CPP29 containing the PRAD1 gene and its 5′‐flanking region split and co‐localized with both Cγ and VH gene probes, thus spanning the breakpoint. In CLL and SBLL patients, donor chromosomes were other than chromosomes 2, 11, 18 and 19, suggesting the involvement of a novel oncogene(s) in the pathogenesis of these diseases. Interphase FISH rapidly detected 14q32.33 translocation, t(11;14) and t(14;18) in B‐cell malignancies with low mitotic activity at the single‐cell level, facilitating the correlation of the molecular features of these translocations with clinical characteristics. Int. J. Cancer 72:31–38, 1997.

Analysis of surface antigen expression of human immunoglobulin‐secreting cells: phenotypic heterogeneity in normal counterparts of myeloma cells

Human myeloma cells are malignant counterparts of plasma cells which represent the most differentiated B cells. Myeloma cells are, however, heterogeneous in their surface antigen expression (Katagiri et al, 1984, 1985), which may reflect that normal plasma cells have a spectrum of differentiation. To test this hypothesis, immunoglobulinsecreting cells (ISC) of non‐neoplastic nature were studied with regard to their surface antigen expression by using a combination of reverse haemolytic plaque assay and complement‐dependent cytolysis. Non‐neoplastic ISC were found to have a broad spectrum of differentiation stages from the immature type of CD20+, HLA‐DR+, CD38+ in the peripheral blood to the mature type of CD20–, HLA‐DR–, CD38+ in the bone marrow. In patients with polyclonal B cell activation (PBA), ISC showed a more immature antigen expression in comparison with ISC in normal controls or patients without PBA. The surface antigen development of ISC was clearly demonstrated throughout the stages in the analysis of mitogen‐induced ISC in vitro. No significant difference in the surface phenotype of ISC was found among heavy chain classes. Thus, non‐neoplastic ISC show a spectrum of differentiation similar to that of myeloma cells, depending on the site where ISC are located, and on the degree of PBA in vivo.

Formation of Tubuloreticular Inclusions in Mitogen-Stimulated Human Lymphocyte Cultures by Endogenous or Exogenous Alpha Interferon

Tubuloreticular inclusions (TRI) were induced in normal blood lymphocytes after incubation with Staphylococcus aureus Cowan 1 (STA), but they were not induced by pokeweed mitogen (PWM), as we reported previously. TRI were also induced in Raji cells when grown in the medium of STA culture. Alpha-interferon (alpha IFN) was detected only in the medium of STA culture and not in PWM culture. The cells of PWM cultures formed TRI when exposed to various concentrations of human leukocyte alpha IFN. The incidences of TRI-positive cells in the presence of 50-500 IU/ml of alpha IFN were 3-5% on day 2 and increased to 10% on day 7. On days 5-7 of the PWM cultures, plasmacytoid cells containing TRI were seen not infrequently. In the presence of a high concentration of alpha IFN (10,000 IU/ml), which was sufficient to inhibit cell growth and differentiation, the growth of the TRI region was not altered and the incidence of TRI-positive cells was 9% on day 2 and increased to 15% on day 7. Our observations suggest that the TRI formation in STA culture is attributable to the alpha IFN produced endogenously by STA-stimulated cells and that some relationship might exist between the incidences of TRI-positive cells in these mitogen-stimulated cultures and the biologic functions of IFN.

Acute promyelocytic leukemia with an intracerebral mass and meningeal involvement after treatment of non‐Hodgkin's lymphoma

After a 3‐month leukopenic phase, a patient developed hematologic pictures of acute promyelocytic leukemia (APL). Treatment with chemotherapy and radiotherapy had been given 10 months previously for a non‐Hodgkins lymphoma (NHL) of diffuse large cell type arising in the Waldeyers ring. A computed tomographic scan demonstrated a high density mass in the left frontal cerebrum, which was enhanced uniformly by the contrast material. Cytocentrifuge examination of cerebrospinal fluid also showed an excess of promyelocytes. From these observations, the mass was considered to be an infiltration of leukemic cells. The intracerebral mass and meningeal involvement were resolved concomitantly with a hematologic remission of APL after intrathecal injection of cytosine arabinoside and systemic combination chemotherapy. This is a particular case of APL previously undescribed, representing an unusual presentation with intracerebral APL mass as well as a rare posttherapeutic APL following NHL. Cancer 59:94–98, 1987.

Helper T-cell lymphoma with marked plasmacytosis and polyclonal hypergammaglobulinemia a case report

A case of malignant lymphoma with a helper activity of neoplastic cells is reported. On admission, a significant number of plasma cells of polyclonal nature were seen in the peripheral blood, and polyclonal hypergammaglobulinemia was seen. The biopsied lymph node showed poorly differentiated lymphocytic lymphoma with marked proliferation of plasma cells. At the terminal stage, the patient became leukemic in contrast with the disappearance of plasma cells from the peripheral blood. Although the leukemic cells failed to form sheep erythrocyte rosettes, they were considered to be of T‐cell origin morphologically. Cytochemically, they had a “dot”‐like pattern of α‐naphtyl acetate esterase and acid phosphatase activity. Ultrastructurally, they had highly convoluted nuclei, and cytoplasmic clustered dense bodies. They showed marked helper activity on pokeweed mitogen‐induced B‐cell differentiation in vitro. This case may provide a novel view concerning the cause of hypergammaglobulinemia induced by lymphoproliferative disorders.

Cefozopran, meropenem, or imipenem–cilastatin compared with cefepime as empirical therapy in febrile neutropenic adult patients: A multicenter prospective randomized trial

We conducted an open-label, randomized study to evaluate the clinical efficacy of cefozopran, meropenem or imipenem-cilastatin using cefepime as a control in febrile neutropenia (FN) patients. Three hundred and seventy-six patients received cefepime, cefozopran, meropenem or imipenem-cilastatinas initial therapy for FN. The primary endpoint was the non-inferiority of response rates including modification at day 7 in cefozopran, meropenem or imipenem-cilastatin patients compared with cefepime in the per-protocol population (delta = 10%). The response rates for cefozopran, meropenem and imipenem-cilastatin were not significantly different compared with cefepime (cefozopran: 54/90 (60%), meropenem: 60/92 (65%), and IPM/CS: 63/88 (72%) versus cefepime: 56/85 (66%) (p = 0.44, 1.0 and 0.51, respectively)), and the differences in treatment success for cefozopran, meropenem and imipenem-cilastatin compared with cefepime were -5.9% (95% confidence interval (CI): -20.1-8.4), -0.7% (95% CI: -14.6-13.3), and 5.7% (95% CI: -8.1-19.4), respectively. The same tendency was seen in the modified intention-to-treat population. Based on the evaluation of initial drug efficacy performed on days 3-5, there was no significant difference between the four drugs. In the subgroup with an absolute neutrophil count ≤ 100 × 10(6)/L for longer than seven days, there was significantly better efficacy in the carbapenem arm compared to 4th generation beta-lactams (52% versus 27% at days 3-5, p = 0.006, and 76% versus 48% at day 7, p = 0.002). Our results suggest that the effects of these four drugs as empiric therapy were virtually the same for adult FN patients, although non-inferiority was shown only in imipenem-cilastatin compared with cefepime (clinical trial number: UMIN000000462).