Summary:initially reported by Stiff et al 1 in 1983. Makino et al 2 modified this method and evaluated its clinical use for peripheral blood stem cell transplantation (PBSCT) in 1991. A simplified method for cryopreservation at −80؇C of peripheral blood stem cells (PBSC) has been increasThey reported that rates of CFU-GM remained at more than 70% during 18 months of cryopreservation and that rapid ingly used for autologous PBSC transplantation in Japan. Although this method, using 6% hydroxyethyl and sustained trilineage engraftment was obtained in 10 patients who received marrow-ablative chemotherapy and starch (HES) and 5% dimethyl sulfoxide (DMSO) as a cryoprotectant without rate-controlled freezing, has sevautotransplantation of PBSC cryopreserved by this method. While this simplified method has several advantages eral advantages over the conventional method using 10% DMSO with rate-controlled freezing, little is such as being a quick and inexpensive procedure and less clumping of the thawed cells compared with the convenknown about effects of long-term cryopreservation for years and thawing process on hematopoietic progenitional method using rate-controlled freezing with 10% DMSO and storage in liquid nitrogen, 2 little is known about tors. We examined the recovery rates of BFU-E and CFU-GM in sample tubes cryopreserved by the simplithe influence of long-term cryopreservation for years and of the thawing process on frozen-thawed hematopoietic fied method under various conditions as follows: (1) long-term storage for 1-5 years; (2) DMSO exposure stem cells. In order to assess the clinical efficacy of this simplified for 1 h after rapid thawing; and (3) thawing at a lower temperature other than 37؇C. In our study, we found method, we investigated effects of long-term cryopreservation (1-5 years), exposure of frozen-thawed cells to that the recovery rates of BFU-E and CFU-GM were not affected by the length of cryopreservation period; DMSO, and slow thawing at room temperature on hematopoietic activity of PBSC when they were cryopreserved by they remained at more than 70% on average for 16-61 months. In our hands, a 1-h exposure to DMSO after this method. rapid thawing was not toxic for hematopoietic progenitors. Furthermore, there was no significant difference in the recovery rates of BFU-E and CFU-GM between Materials and methods thawing at 37؇C and 20؇C. These observations indicate that PBSC cryopreserved for at least 5 years by the simPreparation of peripheral blood mononuclear cells plified method can be used clinically without losing hematopoietic activity, and suggest that hematopoietic Peripheral blood mononuclear cells (PBMNC) were collected by apheresis from patients with hematologic maligactivity of the thawed PBSC may be unaffected when PBSC are infused slowly within 60 min or even when nancies in remission during hematologic recovery from consolidation chemotherapy with or without G-CSF. Aph-PBSC are thawed gradually at room temperature.

Allogeneic peripheral blood stem cell transplantation from an HLA-identical sibling was performed for a 38-year-old male with refractory acute myeloblastic leukemia. The patient was conditioned with total body irradiation (TBI) and high-dose cytosine arabinoside (Ara-C). G-CSF (300 microg/body) was started for priming of residual leukemic cells 24 hr before the beginning of TBI (day -9). However, intolerable generalized bone pain appeared shortly after the start of first dose of G-CSF, and persisted for 3 days in spite of the cessation of G-CSF. Posttransplant hematopoietic engraftment was very rapid. Bone marrow biopsy specimens on day 14 and 30 showed typical bone marrow necrosis histologically. This is the first case of bone marrow necrosis during administration of G-CSF, and our experience suggests that PBSC could repopulate hematopoiesis in spite of severe bone marrow necrosis.

Summary:CD56 + angiocentric lymphoma has currently been recognized as a distinct clinical entity which is the prototype of the putative NK cell lymphomas. A 16-year-old Japanese girl with advanced CD56 + angiocentric lymphoma received high-dose chemotherapy supported with syngeneic peripheral blood stem cell transplantation (PBSCT). Prior to syngeneic PBSCT, she received six cycles of conventional chemotherapy before transplantation, resulting in a partial response. PBSC were mobilized with granulocyte colony-stimulating factor (G-CSF) and collected from her identical twin. High-dose cyclophosphamide, MCNU, etoposide, and carboplatin were used for pretransplant conditioning. Syngeneic PBSCT was well tolerated. She achieved complete remission and is now surviving in continuous complete remission for more than 30 months after syngeneic PBSCT. Thus, marrow-ablative chemotherapy facilitated by autologous or allogeneic PBSCT should be considered as part of the primary therapy for poor prognosis NK cell lymphomas.

Summary. In two-thirds of patients with splenic lymphoma with villous lymphocytes (SLVL) a small amount of M-protein can be detected in association with the presence of plasma cells in the peripheral blood (PB) and/or bone marrow (BM). However, it is not known whether lymphoma cells and plasma cells originate from the same clone. In this report we describe a case of SLVL which was characterized by the presence of marked monoclonal gammopathy (IgG-k 90 g/l) and increased plasma cells in the BM. In an attempt to elucidate the origin of lymphoma cells and plasma cells, we performed morphological, cytogenetic and molecular studies on PB mononuclear cells (PBMNC) without plasma cells and BMMNC containing 10% plasma cells from this patient.Immunofluorescence showed that lymphoma cells and plasma cells were positive for cytoplasmic g heavy and k light chains. Well-developed endoplasmic reticulum was observed in the cytoplasmic organelles of PBMNC using an electron microscope. The mean IgG concentration in the 3 d supernatant cultures of PBMNC was 374 Ϯ 24 mg/l. More than 50% PBMNC differentiated into plasmacytoid cells in 6 d of liquid culture with IL-3 and IL-6. Analysis by twocolour FISH revealed that karyotypic abnormalities of monosomy X and trisomy 17 existed simultaneously in both lymphoma cells and plasma cells. JH gene rearranged bands from PBMNC and BMMNC by Southern blot hybridization were identical, whereas DNAs from PBMNC failed to hybridize with the Cm probe.These observations strongly suggest that lymphoma cells and plasma cells originate from the same clone, and that plasma cells, as well as lymphoma cells, which have undergone class switch recombination, could produce IgG type M-protein in this case.

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