Hitoshi Ohno

Stimulation of CD30 in anaplastic large cell lymphoma leads to production of nuclear factor-κB p52, which is associated with hyperphosphorylated Bcl-3

Anaplastic large cell lymphoma (ALCL) and Hodgkin lymphoma (HL) express CD30 at high levels, but stimulation of this molecule has been reported to induce contradictory effects. To elucidate the molecular mechanism of CD30‐mediated apoptosis of ALCL, we compared the gene expression profiles of t(2;5)(p23;q35)‐positive ALCL with those of HL altered by CD30 agonistic stimulation. The results showed that BCL3, the high‐level expression of which in ALCL was previously reported, was further upregulated in response to CD30 stimulation, along with several pro‐apoptotic genes. Bcl‐3 protein was present as an intermediate phospho‐form in the resting‐state ALCL, becoming hyperphosphorylated (Bcl‐3P) upon stimulation. We next found that the stimulation promoted de novo synthesis of the nuclear factor (NF)‐κB2/p100 precursor as well as processing to p52, and a series of immunoprecipitation and western blotting analyses consistently showed association of Bcl‐3P with p52 in CD30‐stimulated ALCL. An electrophoretic mobility shift assay revealed the induction of κB binding activity of the p52 homodimer, and nuclear colocalization of Bcl‐3 and p52 was demonstrated in anaplastic lymphoma kinase‐positive ALCL tumor tissues by immunohistochemistry. As Bcl‐3 can act as an antirepressor or transactivator or both, we propose that the (p52)2/Bcl‐3P ternary complex, which is specifically induced in CD30‐stimulated ALCL, can modulate expression of apoptosis‐related genes regulated by NF‐κB, thereby accounting for CD30‐mediated apoptosis of ALCL. (Cancer Sci 2005; 96: 487–497)

A contact investigation of the transmission of Mycobacterium tuberculosis from a nurse working in a newborn nursery and maternity ward

A nurse working in a newborn nursery and maternity ward developed 3+ smear-positive lung tuberculosis. The hospital infection control committee, in collaboration with the local public health and welfare center, conducted a contact investigation. The infection period was defined as April to August 2006. The investigation included 109 infant and mother pairs, 28 children aged under 10 years and their guardians, 62 coworkers, and 63 household visitors to the ward. Tuberculosis infection in infants and children aged under 5 years was primarily determined by tuberculin skin test (TST), while subjects aged 5 years or more were tested using QuantiFERON-TB Gold (QFT). The first investigation, in August 2006, was conducted in all subjects, and the second investigation, in October 2006, targeted selected subjects. No infants were TST-positive. Two children aged 1 year or under, vaccinated with bacillus Calmette-Guérin, were positive for TST, as determined by the criteria of the Japan Anti-Tuberculosis Association; however, other tests for tuberculosis were negative. Of the 13 QFT-positive adult subjects, 1 mother and 2 coworkers could have become infected with Mycobacterium tuberculosis through exposure to the index nurse. Fifty-four infants and 6 children underwent “window-period” prophylaxis, and 4 adults completed 6-month prophylactic treatment with isoniazid. To date, no secondary cases of tuberculosis disease have occurred.

Phase II study of ABVd therapy for newly diagnosed clinical stage II–IV Hodgkin lymphoma: Japan Clinical Oncology Group study (JCOG 9305)

Although ABVD (doxorubicin, bleomycin, vinblastine and dacarbazine) therapy has been regarded as a standard of care for advanced-stage Hodgkin lymphoma (HL) since 1992, there has been no prospective data of ABVD therapy in Japan. To investigate the efficacy and safety of ABVd therapy with the lower dose of dacarbazine (250xa0mg/m2) in patients with newly diagnosed stage II–IV HL, Lymphoma Study Group of Japan Clinical Oncology Group conducted a phase II study. The primary endpoints were complete response rate (%CR) and progression-free survival (PFS). A total of 128 patients with age less than 70xa0years were enrolled and received 6–8 cycles of ABVd followed by radiation to initial bulky mass. The %CR in 118 eligible patients was 81.4% [95% confidence interval (CI) 73.1–87.9%]. Major toxicity was grade 4 neutropenia (45.3%). Grade 3 nausea/vomiting was the most frequent non-hematological toxicity (10.9%). Transient grade 4 constipation, infection (abscess), hypoxemia and hyperbilirubinemia were observed in 4 patients. No treatment-related death was observed. PFS and overall survival at 5xa0years were 78.4% (95% CI 70.9–85.9%) and 91.3% (95% CI 86.1–96.5%), respectively. In conclusion, ABVd is effective in Japanese patients with stage II–IV HL with acceptable toxicities (UMIN-CTR Number: C000000092).

Rituximab alone was effective for the treatment of a diffuse large B-cell lymphoma associated with hemophagocytic syndrome

We report here the case of a 63-year-old man who had a diffuse large B-cell lymphoma associated with hemophagocytic syndrome (HPS). The lymphoma involved the spleen, bilateral adrenal glands, and paraaortic lymph nodes of the abdomen. In both the bone marrow and lymph nodes, hemophagocytosis was evident, and the laboratory findings were consistent with HPS. The lymphoma cells showed a CD4+, CD5+, CD10−, CD19+, CD20+, CD25+ and surface immunoglobulin µα/κ+ immunophenotype. The patient was unintentionally treated with rituximab alone, resulting in complete resolution of the lymphomatous lesions as well as the features of HPS in response to the initial two doses of rituximab, although he developed gastric hemorrhage requiring vigorous resuscitation. After the completion of eight doses of rituximab, the patient remains free of disease with an excellent performance status.

Reappraisal of BCL3 as a Molecular Marker of Anaplastic Large Cell Lymphoma

The BCL3 gene was initially discovered through its involvement in a recurring translocation, t(14;19)(q32;q13), which is found in some patients with B-cell chronic lymphocytic leukemia (B-CLL). The translocation leads to the juxtaposition of BCL3 to the immunoglobulin heavy chain gene locus, resulting in high-level expression of the BCL3 transcript. The Bcl-3 protein includes 7 tandem copies of the ankyrin repeat element in the central domain, a structure that is characteristic of the IκB family of inhibitors of the nuclear factor κB transcription factors. Anaplastic large cell lymphoma (ALCL) is a subtype of aggressive non-Hodgkin’s lymphoma that is characterized by expression of CD30 and the NPM/ALK chimeric protein, which is generated by t(2;5)(p23;q35). We compared the gene expression profiles of ALCL with those of another CD30+ neoplasm, Hodgkin’s disease (HD), and found that BCL3 is expressed at higher levels in ALCL than in HD. A comparison by real-time polymerase chain reaction assay revealed that t(2;5)+ ALCL expresses a high level of BCL3 messenger RNA relative to the levels expressed in other hematologic tumors, and the level in ALCL is comparable to or even higher than that in t(14;19)+ B-CLL. An immunohistochemical analysis of ALCL tumor tissues showed that the lymphoma cells exhibited strong nuclear staining by a monoclonal antibody against Bcl-3. We suggest that Bcl-3 sequestrates the (p50)2 homodimer to the nucleus and that the κB sites are occupied by the (p50)2/Bcl-3 ternary complex. Future studies should identify the relationships among the 3 independent molecules (ie, NPM/ALK, CD30, and Bcl-3) that are activated in t(2;5)+ ALCL.

Epstein-Barr virus-positive diffuse large B-cell lymphoma carrying a t(9;14)(p13;q32) translocation

We, herein, report a 75-year-old man with lymphoma who initially presented with disseminated disease involving the lung, followed by temporal regression, and finally died of disease progression. Lymph-node biopsy showed a morphology of diffuse large B-cell lymphoma (DLBCL), containing CD30+ Reed–Sternberg-like cells. The lymphoma cells were stained by in situ hybridization (ISH) for Epstein–Barr virus (EBV)-encoded RNA, and the presence of the EBV genome was confirmed by the polymerase chain reaction. A cytogenetic study showed that the lymphoma cells carried a t(9;14)(p13;q32) translocation, and rearrangement of the PAX5 gene was determined by fluorescence ISH using a split signal probe. This case report is the first to identify t(9;14)(p13;q32) in EBV+ DLBCL of the elderly, which was very recently listed among subtypes of DLBCL.

Polyclonal Proliferation of Plasma Cells Associated with Marked Hypergammaglobulinemia in an Elderly Patient

We describe an 89-year-old woman who presented with prominent plasmacytosis mimicking plasma cell leukemia. The apparent serum M-protein level of >7 g/dL of γ mobility was revealed to be a polyclonal increase of immunoglobulins. The plasma cells in the peripheral blood expressed polyclonal surface/cytoplasmic immunoglobulins as well as CD19, CD30, CD38, and CD138 antigens but lacked CD10, CD20, CD25, and CD56. The bone marrow plasma cells showed the CD45+, CD19+, CD56-, MPC-1-/+, and CD49e- immunophenotype, which was in clear contrast with the immunophenotypes of the neoplastic myeloma cells. Abdominal lymphadenopathy, splenomegaly, and a high level of soluble interleukin 2 receptor may have been reflections of an underlying lymphoproliferative disorder, potentially leading to the polyclonal proliferation of plasma cells.

Characterization of de novo diffuse large B-cell lymphoma with a translocation of c-myc and immunoglobulin gènes

The characteristics of de novo diffuse large B-cell lymphoma (DLBCL) with translocation of c-myc and immunoglobulin (Ig) genes (c-myc/Ig DLBCL), were investigated in 13 cases of c-myc/Ig DLBCL. Immunohistochemistry revealed five cases were positive for CD10 and BCL6 expression (CD10(+)/BCL6(+)), five cases of CD10(-)/BCL6(+)/MUM1(-), one case of CD10(-)/BCL6(+)/MUM1(+) and two cases of CD10(-)/BCL6(-)/MUM1(+) expression, indicating 10 cases of germinal center B-cell DLBCL and three cases of non-germinal center B-cell DLBCL. Ongoing mutation of the Ig heavy chain gene variable region (IgH-V) was found in two cases with CD10 and BCL6 expression and one case showing CD10(-)/BCL6(+)/MUM1(-) expression. These three cases of ongoing mutation of the IgH-V gene did not express BCL2, unlike those without ongoing mutation. These results suggest a heterogeneous immunophenotype and genotype for c-myc/Ig DLBCL, with CD10(-)/BCL6(+)/MUM1(-) cases the most frequent.

Long-term response to maintenance treatment with rituximab in CD20+ multiple myeloma

The human–mouse chimeric monoclonal antibody against the CD20 antigen, rituximab, has been used in a number of clinical settings in patients with B-cell tumors. Maintenance treatment of rituximab after successful induction treatment, either as 4-weekly infusions every 6 months or as a single infusion every 2–3 months, has been reported to prolong the progression-free interval in indolent B-cell lymphomas, even though clear evidence of improvement in overall survival is lacking [1]. CD20 is expressed in a fraction of multiple myeloma (MM) [2,3]; however, the activity of rituximab against CD20þ MM is undetermined and controversial [4]. A literature review of case reports and small phase 2 clinical trials shows that the response to rituximab has been variable, ranging from rapid disease progression to stabilization of the disease, and even a partial response [5–9]. On the other hand, few cases treated with a maintenance schedule have been reported to date [5]. Here, we describe a case of CD20þ MM, which showed a long-term response to maintenance treatment with rituximab. A 72-year-old woman presented to our hospital with anemia. The hemoglobin level 3 years before presentation was 10.2 g/dL, which progressively fell to 5.4 g/dL. The white cell count was 3,700/ml, and that of platelets was 1336 10/ml. The blood chemistry values included: total protein, 6.9 g/dL; albumin, 3.9 g/dL; lactate dehydrogenase, 177 IU/L; creatinine, 1.40 mg/dL; calcium, 8.1 mg/dL. Protein electrophoresis revealed an M-spike within the g globulin region, and immunoelectrophoresis confirmed that the monoclonal immunoglobulin (Ig) in the serum was labeled by IgG and l-chain anti-sera and the l light chain was excreted in the urine. The serum levels of Ig were: IgA, 29 mg/dL; IgG, 2,262 mg/dL; IgM, 31 mg/dL. The level of b2microglobulin in the serum was 7.67 mg/L and that in the urine was 4,918 mg/L. There was no lytic lesion on skeletal examination. The bone marrow was normocellular and contained 20.6% myeloma cells showing a small mature plasma cell morphology (Figure 1A, left). The cells were CD13, CD19, CD20þ, CD38þ, CD45, CD138þ, and HLA-DR, as determined by flow cytometry; the percentage of CD20þ cells was 60.5%. Immunohistochemistry of the histologic sections prepared from the bone marrow aspirates confirmed that the myeloma cells expressed IgG/l in the cytoplasm and CD38 and CD20 on the cell surface (Figure 1A, right). Cyclin D1 expression was negative. Cytogenetic analysis revealed a normal female karyotype, and fluorescence in situ hybridization for t(11;14)(q13;q32) translocation was negative. The patient was diagnosed with MM categorized as stage III disease using both the Durie-Salmon and International Staging Systems due to the severity of anemia and high level of serum b2microglobulin [10,11]. The patient was initially treated with melphalan plus prednisone; however, she declined further treatment with cytotoxic drugs. In order to obtain consent for treatment with rituximab, we informed her of: (1) the rationale of the treatment,

The duration of functioning of a subcutaneous implantable port for the treatment of hematological tumors: a single institution-based study

BackgroundAlthough subcutaneous implantable ports have been indicated as venous access for chemotherapy, these devices have not been used routinely for hematological tumors.MethodsBetween May 2006 and April 2009, 39 ports were implanted in 37 patients with hematological tumors and 16 ports were implanted in 14 patients with nonhematological tumors. The patients were treated with standard/first-line and/or salvage/second-line or greater chemotherapy, and were prospectively followed until port removal, death, or the end of the study.ResultsThirty-five (96%) patients with hematological tumors developed grade 4 hematological toxicity, while 1 (7%) patient with nonhematological tumors showed grade 4 neutropenia. The actual duration of the port in situ ranged from 14 to 719 days (mean, 271.4 days) in the hematology group, and from 50 to 955 days (mean, 419.5 days) in the nonhematology group (Pxa0=xa00.039). The Kaplan–Meier-estimated median duration of port in situ in the hematology group was 364xa0days, which was significantly shorter than that in the nonhematology group (Pxa0=xa00.009). When patient death and port removal for the end of treatment were censored, the rate of port functioning at 1xa0year was estimated to be 83% in the hematology group. Bloodstream infection (BSI) occurred in 7 patients with hematological tumors and in 1 patient with metastatic colorectal cancer; however, microbiological confirmation that the implantable port was the source of the BSI was inconclusive.ConclusionThe duration of port functioning in patients with hematological tumors was comparable to that in patients with nonhematological tumors. The higher rate of BSI in the hematology group was primarily attributable to profound neutropenia.