Hirotaka Nakahashi

Circulating plasmacytoid dendritic cells in patients with primary and Helicobacter pylori-associated immune thrombocytopenia.

Objectives:  Immune thrombocytopenia (ITP) is an autoimmune disorder characterized by the production of autoreactive antibodies against platelet antigens. Although dysfunction of multiple aspects of cellular immunity is considered to be important in the pathogenesis of ITP, it has not been clarified which cell types play a principal role.

Characterization of immunoglobulin heavy and light chain gene expression in chronic lymphocytic leukemia and related disorders.

The incidence of chronic lymphocytic leukemia is low in the Japanese population compared with populations in western countries, suggesting a role for genetic factors in the occurrence of this disease. We have previously shown that chronic lymphocytic leukemia in Japan rarely expresses the immunoglobulin heavy chain variable region (IGHV) 1‐69 gene (1 out of 43 patients, 2.3%), which is a gene most commonly expressed in chronic lymphocytic leukemia cases from western countries. In the current study, we extended the previous study by examining immunoglobulin heavy chain and light chain gene expression in 80 Japanese patients with chronic lymphocytic leukemia and in 52 Japanese patients with other leukemic chronic lymphoproliferative disorders. IGHV1‐69 gene expression was again quite low in our cohort, found in only two patients: one with chronic lymphocytic leukemia and the other with splenic marginal zone lymphoma. The IGHV4‐34 gene was most frequently expressed in chronic lymphocytic leukemia (27.5%), whereas it was rarely found in leukemic chronic lymphoproliferative disorders (7.7%, P = 0.005). There was also a significant difference in the expression of IGLV3‐21 between chronic lymphocytic leukemia and leukemic chronic lymphoproliferative disorders (29.4 vs 4.8%, P = 0.018). The IGLV3‐21 gene in the majority of chronic lymphocytic leukemia cases was associated with homologous complementarity determining region 3 sequences. Recent studies identified subsets of cases expressing almost identical B‐cell receptors. We found that two patients with chronic lymphocytic leukemia and the patient with splenic marginal zone lymphoma expressed IGHV4‐39/IGKV1‐39 and IGHV1‐69/IGKV3‐20, respectively, which belong to these subsets. (Cancer Sci 2009)

Autologous Peripheral Blood Stem Cell Transplantation to Treat CHOP-Refractory Aggressive Subcutaneous Panniculitis-Like T Cell Lymphoma

his right femur that had swelled for 3 months. A physical examination revealed cutaneous indurations on his lower abdomen, right buttock and right femur. Multiple nontender subcutaneous nodules (1–2 cm in diameter) were evident on his face, back and upper extremities. A nontender right inguinal lymph node (1.5 cm) was palpable. Blood chemistry demonstrated lactate dehydrogenase, 832 IU/l (normal: ! 229 IU/l), and soluble interleukin-2 receptor, 1,909 IU/ml (normal: 220–530 IU/ml). A bone marrow examination revealed normocellular marrow without any lymphoma cell involvement and hemophagocytosis. Soft tissue enlargement of the lower abdomen and right femur ( fig. 1 a) were evident on computed tomographic images and 18 F-fluorodeoxyglucose positron emission tomography images showed that these regions were hypermetabolic. The maximum standardized uptake value was 3.4 in the right inguinal lymph node. A skin biopsy from the right femur showed infiltration of the subcutaneous fatty tissue by atypical medium to large lymphocytes with irregular nuclei rimming around fat vacuoles ( fig. 2 a, b). Immunohistochemical studies revealed that the phenotype of these cells was CD3+, CD4–, CD8+ ( fig. 2 c), CD20–, CD30–, CD45RO+ and CD56–. Cytotoxic molecules, namely granzyme B and T Subcutaneous panniculitis-like T cell lymphoma (SPTCL) is a rare form of postthymic cytotoxic T cell lymphoma that represents ! 1% of all non-Hodgkin lymphomas [1, 2] . The clinical course of SPTCL ranges from mild to progressive and fatal despite aggressive chemotherapy. Treatment strategies for SPTCL remain controversial because of its rarity. Anthracycline-based regimens, cyclophosphamide (CPM), doxorubicin, vincristine and prednisone (CHOP) or CHOP-like combinations have been used most frequently for patients with aggressive disease, but the overall outcome remains poor. The presence of hemophagocytic syndrome (HPS) and CD56 expression by tumor cells are associated with inferior overall survival [2–5] . We describe here a patient with CHOP-refractory aggressive SPTCL despite being HPS and CD56 negative. He achieved complete remission after undergoing an acute lymphoblastic leukemia (ALL) regimen followed by autologous peripheral blood stem cell transplantation (auto-PBSCT) with a conditioning regimen comprising total body irradiation and melphalan. Three years later, he remained in complete remission without treatment. The patient, a 22-year-old Japanese male, presented with persistent high fever and subcutaneous nodules on Received: March 3, 2009 Accepted after revision: March 18, 2009 Published online: June 29, 2009

Flow cytometric detection of human telomerase reverse transcriptase (hTERT) expression in a subpopulation of bone marrow cells

Telomerase activity has been found in most common cancers, thus indicating that telomerase detection may be a useful marker in cancer diagnosis. The telomeric amplification protocol (TRAP) assay and RT-PCR are customarily used to detect telomerase activity and the expression of the associated genes in cells. However, these methods do not provide any information about telomerase activation at an individual cell level. To analyze cells separately, those cells have to be isolated by sometimes complicated method. The immunohistochemical detection of human telomerase reverse transcriptase (hTERT) is useful to detect telomerase positive cells in a background of non-cancerous cells. A method has been developed for the detection of intranuclear hTERT protein, in a subpopulation of hematopoietic cells, using concurrent staining of a cell surface antigen and multicolor flow cytometry. Only mouse monoclonal anti-hTERT antibody demonstrated the specific positivity in immunocytochemistry and immunofluorescent flow cytometry. Human leukemia and myeloma cell lines showed 100% positivity, whereas normal neutrophils showed 0% positivity. hTERT expression was analyzed in hematopoietic precursor cells of bone marrow samples using concurrent staining of surface CD34 antigen and intracellular hTERT protein and multi-parameter flow cytometry. CD34 positive cells demonstrated higher expression of hTERT than CD34 negative cells. A quick, easy and sensitive assay for determining the hTERT protein expression has been developed. Using this method and the multi-parameter nature of flow cytometry and its ability to identify cellular subpopulations will provide a better understanding of the mechanisms regarding the activation of telomerase.

Hairy Cell Leukemia-Related Disorders Consistently Show Low CD27 Expression

In Japan, typical hairy cell leukemia (HCL) is rare, and HCL-Japanese variant (HCL-JV) is more common. Hairy B-cell lymphoproliferative disorder (HBLD) is another unusual disorder of polyclonal B-lymphocytosis of hairy cell appearance. In the present study, we analyzed the clinical features of 3 patients with HCL, 3 with HCL-JV, and 3 with HBLD. All HBLD patients had the DRB1*04 allele. As compared with other B-cell lymphoproliferative disorders, CD27 expression on B cells was significantly lower in all patients, ranging from 0.3% to 23.4%. Our results suggest that low CD27 expression may be a distinct feature of these HCL-related disorders.

Abstract 2147: Molecular allelokaryotyping to identify mechanism of development of Asian chronic lymphocytic leukemia

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Chronic lymphocytic leukemia (CLL) is a common hematological malignancy in western countries. However, this disease is very rare in Asian countries. It is not clear whether the mechanisms of development of CLL in Caucasian and Asian are same. To answer this question, we compared genomic abnormalities in Asian and Caucasian CLL using single-nucleotide-polymorphism genomic microarray (SNP-chip). We analyzed 75 cases of Japanese CLL (Binet stage A: 51 cases, B: 8 cases, C: 16 cases, unknown: 3 cases) using 250-k GeneChip from Affymetrix. We compared these results to 56 cases of Caucasian CLL. Asian CLL had 4 common genomic abnormalities: deletion of 13q14.3 (36 cases, 49 %), trisomy 12 (23 cases, 30 %), abnormalities of 11q (9 cases, 13 %: 7 deletions and 2 uniparental disomy), and abnormalities of 17p (5 cases, 8 %: 4 deletions and 1 uniparental disomy). These 4 common genomic abnormalities were also identified in Caucasian CLL cases. Interestingly, in both racial groups, trisomy 12 and deletion of 13q14.3 were mutually exclusive (p = 0.01). Nevertheless, frequency of trisomy 12 was higher in Japanese CLL cases (23/75 cases, 30 %) than in Caucasian cases (4/56 cases; 7 %) while frequency of deletion of 13q14.3 was comparable in Japanese and Caucasian (49 % and 48 %, respectively). Frequency of deletion of 11q23.1 was similar in both racial groups. Notably, not only did all these cases involve deletion of ATM but always also deleted miR 34b and 34c. Furthermore, one case had a homozygous deletion of 11q22.3 involving caspase 1/4/5 and COP1. Interestingly, all except one CLL case with 11q abnormalities had either deletion or acquired uniparental disomy. Both Asian and Caucasian CLL had a commonly deleted region at 13q14.3 involving miR 15-a and 16-1. Interestingly, one case had two distinct deletions in this region; one involved miR 15-a and 16-1; the other involved Rb1. On close inspection, 36 % of cases with deletion of 13q, had a deletion of Rb1, suggesting that Rb1 might be another target of deletion of 13q. In summary, the new findings of this study are: 1) Genomic abnormalities in Asian and Caucasian CLL were comparable except for a higher frequency of trisomy 12 in Asian CLL; 2)Trisomy 12 and 13q deletions were mutually exclusive suggesting that either lesion can substitute for the other and may involve a common cellular pathway; 3)New CLL genomic abnormalities include deletion of miR 34b and 34c, caspase 1/4/5 and COP1 (11q), and Rb1 (13q). Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2147.

Retrospective analysis of prognostic factors for Waldenstrӧm macroglobulinemia: a multicenter cooperative study in Japan.

Although population-based cancer registries have reported lower incidence of Waldenstrӧm macroglobulinemia (WM) in East Asia than in Western countries, previous retrospective analyses have found the clinical features of WM to be similar in these two populations. To clarify the characteristics of Japanese WM patients, we retrospectively analyzed clinical and laboratory characteristics, treatments, outcomes, and prognostic factors in 93 patients with WM. Based on the Second International Workshop on WM (IWWM-2) criteria, symptomatic WM was found in 73 (78.5%) and asymptomatic WM in 20 (21.5%) of cases examined. The median overall survival (OS) was similar to that in reports from Western countries. Patients receiving treatment regimens including rituximab exhibited significantly better survival than those not given rituximab. Although prognostic factors for WM in Western countries may not apply to Japanese patients, our finding that newly diagnosed WM patients with pleural effusion have a poorer prognosis suggests that this may be a novel predictor of adverse prognosis in symptomatic WM.

A case of myasthenia gravis with transient taste disorders followed by aplastic anemia after thymectomy

A 45-year-old man was admitted to our hospital because of taste disorders in March 2014. He exhibited cervical muscle weakness and left eye ptosis, which responded to Tensilon test, and was diagnosed with myasthenia gravis (MG). He developed aspiration pneumonia and myasthenic crisis, which was treated with intravenous immunoglobulin and steroid pulse therapy. All symptoms disappeared. Oral administration of prednisolone and tacrolimus was started. Chest CT revealed thymoma and extended thymectomy was performed in May 2014. In December 2014, seven months after the thymectomy, hematological examination showed pancytopenia including severe neutropenia. We diagnosed his illness as aplastic anemia (AA). Cyclosporine therapy with transfusion was administerd and led to reticulocyte count recovery. Since May 2015, hemoglobin recovery reached a blood transfusion free period. To our knowledge, this is the first case report with the patient supposed of relationship among taste disorders, AA and thymoma-associated MG.