Background: Skin lesions from graft-versus-host disease (GVHD) show histological features of epidermal cell death with lymphocyte infiltration. Perforin and granzyme B are involved in the process of apoptosis induced by cytotoxic T lymphocytes (CTL). Objective: To elucidate the role of CTL in the mechanism of epidermal injury in GVHD. Methods: We studied immunohistochemical staining for granzyme B and perforin in the skin lesions of 8 patients who developed GVHD after bone marrow transplantation. Results: Granzyme-B-positive lymphocytes were CD8 positive and were observed in the epidermis of 3 out of 6 specimens in acute GVHD, and of 5 specimens of chronic GVHD except for 1 sclerotic type in which it was negative. Perforin-positive lymphocytes were observed in the epidermis of the specimens from 1 acute and 1 chronic GVHD. Conclusions: Granzyme B derived from CTL may be involved in the mechanisms of epidermal injury in GVHD.

The loss of a specific chromosomal region provides a clue to the elucidation of the putative tumor suppressor gene implicated in the pathogenesis and progression of tumors. To delineate the specific region(s) involved in lymphomagenesis, we performed a survey of loss of heterozygosity for 11 polymorphic microsatellite loci scattered on variable chromosome arms. We examined 20 primary lymphoma samples, including both indolent and aggressive B-cell non-Hodgkin's lymphoma (B-NHL) and Hodgkin's disease (HD), and found a significant number of B-NHLs with loss of genetic material on chromosome arm 13q at the RB1 locus (50%; 4 of 8 informative cases for the RB1 locus). To specify the 13q deletion and to narrow the critical deleted region, we examined the same 20 lymphomas by intensive microsatellite mapping analysis using 12 microsatellite markers, mapping from 13q12.3 to 13q14. We confirmed the frequent 13q14 deletion to be in the vicinity of the RB1 locus (50% of the informative NHLs for at least 1 of 12 microsatellite loci; 5 of 10 aggressive NHLs and 2 of 4 indolent NHLs, but none of 6 HDs) and determined a subchromosomal region deleted in lymphoma on 13q14 defined by D13S164-D13S273, which is an overlapped region frequently lost in chronic lymphocytic leukemia. Taken together, our data indicate that the 13q alterations are present in a wide variety of NHLs including both indolent and aggressive B-NHLs, suggesting that loss of genetic material at chromosome band 13q14 may play an important role in the formation or development of a wide variety of mature lymphoid malignancies.

The Ki-JK cell line was established from a patient with an anaplastic large cell lymphoma. Ki-JK expresses the CD30 antigen and the product of the p80NPM/ALK chimeric gene. ML-9 is a myosin light chain kinase inhibitor that induces apoptosis in Ki-JK cells. Anti-CD30 antibody (aCD30Ab) inhibited the ML-9-induced apoptosis. We observed the expression of nuclear factor κB (NFκB) using anti-NFκB antibody and flow cytometry. aCD30Ab augmented NFκB expression in Ki-JK cells. Simultaneously, aCD30Ab induced interleukin-8 (IL-8) secretion. Both antisense oligonucleotides of NFκB and IL-8 cancelled the anti-apoptotic effect of the aCD30Ab-induced anti-apoptotic effect. In the second approach, we examined the expression of p80NPM/ALK with immunocytochemistry. aCD30Ab induced the augmentation of p80NPM/ALK in Ki-JK cells. Therefore, the anti-apoptotic effect of aCD30Ab was mediated by activating NFκB, p80NPM/ALK and IL-8. We conclude that p80 NPM/ALK might be related with malignant transformation or tumor evolution.

American Journal of HematologyVolume 63, Issue 2 p. 102-102 Letters and CorrespondenceFree Access Pure red cell aplasia (PRCA) with thymoma: A possible distinct clinical entity distinct from large granular lymphocyte (LGL) leukemia Michihiko Masuda, Michihiko Masuda Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this authorYukari Arai, Yukari Arai Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this authorTakamitsu Okamura, Takamitsu Okamura Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this authorMakio Wada, Makio Wada Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this authorHideaki Mizoguchi, Hideaki Mizoguchi Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this author Michihiko Masuda, Michihiko Masuda Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this authorYukari Arai, Yukari Arai Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this authorTakamitsu Okamura, Takamitsu Okamura Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this authorMakio Wada, Makio Wada Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this authorHideaki Mizoguchi, Hideaki Mizoguchi Department of Hematology, Tokyo Women's Medical University, Tokyo, JapanSearch for more papers by this author First published: 10 January 2000 https://doi.org/10.1002/(SICI)1096-8652(200002)63:2<102::AID-AJH9>3.0.CO;2-PCitations: 4AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume63, Issue2February 2000Pages 102-102 ReferencesRelatedInformation

To clarify the role of allelic loss on chromosome arm 13q in lymphomagenesis, we performed fluorescence in situ hybridization (FISH) analysis of a total of 43 primary lymphomas, including both indolent and aggressive non-Hodgkin's lymphoma (NHL) and Hodgkin's disease (HD), using the specific probes at RB1 and D13S319 loci on the centromeric portion of chromosome arm 13q. Monosomy at either or both RB1 and D13S319 loci was detected in 15 of 43 (35%) lymphomas (14 of 43 cases at RB1 locus and seven of 43 cases at D13S319 locus); the 13q deletion was frequently detected in the aggressive NHLs (40%; 12 of 30 cases) compared with that in indolent NHL (17%; one of six cases) and a subset of HD (29%; two of seven cases). There are only six cases of 43 which have total monosomy 13q14, all aggressive NHL, 14% of total or 20% of this subgroup. In addition, we analyzed the loss of heterozygosity in 15 of the 43 primary lymphoma samples for several polymorphic microsatellite loci (D13S168, RB1 and D13S272) on the chromosome arm 13q, and confirmed the 13q deletion in four of five cases that were positive on FISH analysis. The subchromosomal region frequently altered in lymphoma on 13q14 is the region around RB1 locus and centromeric to D13S319 locus, which is an overlapped region frequently deleted in chronic lymphocytic leukemia. Together, our data indicate that the 13q alterations are present in a variety of types of lymphoma and occur in a significant proportion of aggressive NHLs, suggesting the possible presence of common candidate gene(s) on the 13q14 region, whose alteration may play an important role in the formation or development of a wide variety of mature lymphoid malignancies.

All-trans retinoic acid (ATRA) is a vitamin A derivative that induces the differentiation of myeloid leukemia cells in vitro and in vivo. Several investigators have recently reported that ATRA downregulates the production of interleukin-6 (IL-6) and the expression of IL-6 receptor (IL-6R) and also inhibits the proliferation of myeloma cells. It has also been reported that myeloma cells express Fas antigen, and in some of these cells apoptosis was induced by treatment with anti-Fas monoclonal antibody (mAb). In the present study, we demonstrated that ATRA increased Fas expression in the human myeloma cell line, U266B1. We observed that both apoptosis induction and growth inhibition were enhanced in cells exposed to a combination of anti-Fas mAb and ATRA compared with cells exposed to either treatment alone. We also examined whether ATRA modulated bcl-2, an anti-apoptosis protein, in U266B1 cells. Flow cytometry analysis revealed that the mean fluorescence intensity of bcl-2 protein was slightly decreased in cells treated with ATRA. These results indicate that in U266B1 cells, combined treatment with anti-Fas mAb and ATRA enhances the induction of apoptosis by modulating the expression of Fas and bcl-2 by ATRA.

CD30 is a member of the tumor necrosis factor superfamily. In this study, we examined the effect of four anti-CD30 (aCD30) antibodies (Abs) on CD30-positive anaplastic large cell lymphoma-derived cell line, Ki-JK. The aCD30 Abs suppressed [3H]thymidine (TdR) incorporation. With a TdT mediated dUTP-biotin nick end labeling method, apoptosis was detected in Ki-JK cells at day 5 after the addition of aCD30 Ab to the culture. Genistein, an inhibitor of protein tyrosine kinase, had no effect on aCD30 Ab-induced apoptosis. The aCD30 Ab simultaneously induced interleukin-8 (IL-8) secretion in the Ki-JK cells. In culture of the Ki-JK cells with aCD30 Ab for 5 days, the IL-8 concentration of the cell free-supernatant increased to 240 +/- 16 pg/ml, though the concentration was < 12.5 pg/ml without aCD30 Ab. In combination with aCD30 Ab, genistein decreased the concentration of IL-8 in day 5 supernatants. Although, doxorubicin and herbimycin-A suppressed [3H]TdR incorporation and induced apoptosis in the Ki-JK cells, they did not induce IL-8 secretion. Only aCD30 Ab-induced apoptosis was accompanied by IL-8 secretion. IL-8 mRNA was not detected in the Ki-JK cells by reverse transcription-polymerase chain reaction assay. IL-8 mRNA was detected 5 days after adding aCD30 Ab to the culture.

Pure red cell aplasia (PRCA) sometimes accompanies thymoma. Herein, we report a PRCA patient with thymoma with a clonal disorder of T cells. A 55-year-old man presented with anemia and anterior mediastinum tumor. The laboratory study revealed hemoglobin 8.2 g/dl; leukocytes 15.8 × 109/L with 76.5% neutrophils, 20.0% lymphocytes, and reticulocytes 0.0%. Bone marrow aspirate smears and biopsy sections revealed normal myeloid and megakaryocyte differentiation and contained no erythroid precursors. We made the diagnosis of PRCA. The size of the lymphocytes was small without any granules in the cytoplasm. The surface marker of peripheral blood mononuclear cells demonstrated increased CD2+, CD3+, CD4−, and CD8+ populations. The mediastinal tumor was resected and a thymoma diagnosed. A monoclonal rearrangement of T-cell receptor (TCR)- β-chain gene was found using Southern blot analysis of the mononuclear cells in both peripheral blood and thymoma. Treatment with prednisolone, thymectomy, and cyclophosphamide exerted no beneficial effect. After initiation of the Cyclosporin A therapy, the patient developed reticulocytosis. This PRCA case seems to present a neoplastic proliferation of CD8+ T cells in peripheral blood and thymus with a monoclonal rearrangement of the TCR- β-chain gene. Am. J. Hematol. 54:324–328, 1997. © 1997 Wiley-Liss, Inc.

A 54-year-old woman with leukocytosis, was referred to our clinic in February 1982. Based on findings of pancytosis, high NAP score, high serum vitamin B12, increase in total red cell volume and splenomegaly, she was diagnosed as having polycythemia vera (PV). Since then, she has been treated with pipobroman, hydroxycarbamide and phlebotomy. Leukocytosis with increase in blastic cells and thrombocytopenia was noted in August 1995, and she was admitted to our hospital. Since the blastic cells were CD10(+)19(+)20(+), she was diagnosed as having acute lymphoblastic leukemia and treated with vincristine and prednisolone, resulting in remission. This case suggests that PV is a disease of multipotent stem cells including those with a lymphoid lineage.

Activated leukocytes and oxygen free radicals have been implicated in the pathogenesis of heart and lung injury after reperfusion and during cardiopulmonary bypass. This study was designed to determine whether leukocyte depletion prevents injury to the heart and lung during cardiopulmonary bypass. Twenty-eight open heart surgeries were performed in this study. In Group F, leukocyte depletion was performed with an LG-6 arterial line filter after aortic declamp (n = 14). Leukocyte depletion was not performed during cardiopulmonary bypass in Group C (n = 14). Thereafter, cardiac and lung function were assessed in the 24 hr after reperfusion. The total catecholamine dose used for 24 hr after reperfusion (r) was 61.9 +/- 13.4 in Group C and 43.9 +/- 19.2 in Group F (p < 0.05). CK-MB at 3 and 6 hr after reperfusion was 65.9 +/- 13.5 and 64.8 +/- 15.8 in Group C and 45 +/- 11.8 and 38 +/- 10.8 in Group F, respectively (p < 0.05). The pulmonary index after reperfusion at 3 and 6 hr was 1.7 +/- 0.5 and 1.3 +/- 0.4 in Group C and 0.7 +/- 0.3 and 0.6 +/- 0.4 in Group F, respectively (p < 0.05). There was significantly better preserved lung function in Group F. In conclusion, leukocyte depletion was significantly effective in preserving heart and lung function during cardiopulmonary bypass.