Anemia of chronic disease (ACD) is commonly observed in chronic inflammation, although its pathogenesis is poorly understood. Hepcidin is thought to be a key regulator in iron metabolism and has been implicated in ACD. Although the induction of hepcidin by an inflammatory cytokine interleukin-6 (IL-6) seems to have been confirmed, it is still controversial whether interleukin-1beta (IL-1beta), also known as an inflammatory cytokine, regulates hepcidin expression. We demonstrated that hepcidin mRNA was upregulated by IL-1beta in human hepatoma-derived HuH-7 cells, particularly at low concentrations of IL-1beta, while high concentrations of IL-6 were needed for the upregulation of hepcidin mRNA. Therefore, IL-1beta might be more important for the upregulation of hepcidin in physiological conditions than IL-6. Although IL-1beta induces IL-6 production in hepatocytes, our data indicate that the effect of IL-1beta on hepcidin expression is independent from that of IL-6. In conclusion, IL-1beta might have an important role in ACD.

Imatinib mesylate is a specific inhibitor of BCR-ABL tyrosine kinase, which is now widely used for the treatment of chronic myeloid leukemia (CML) with a high efficacy. Although severe hepatic injury caused by imatinib mesylate is rare, such a side effect may force patients to discontinue taking imatinib mesylate. In the present paper, we report on the case of a 51-year-old woman with CML who experienced hepatic injury with severe hyperbilirubinemia caused by imatinib mesylate. The findings from a liver biopsy specimen and her clinical course suggested the hepatic injury to presumably have been caused by an allergic mechanism. The co-administration of prednisolone was thus tried, and she has been able to continue imatinib mesylate administration without any liver dysfunction and finally was able to obtain a complete cytogenetic response.We therefore recommend that prednisolone should be tried when severe hepatic injury caused by imatinib mesylate is observed, since it might enable such patients to continue imatinib mesylate treatment and thereby improve the prognosis in such cases.

Cancer immunotherapy using heat shock protein (HSP) derived from autologous tumor requires cluster of differentiation (CD)4+ as well as CD8+ T-cells for the prolongation of patient survival, suggesting that a humoral immune response through CD4 + T-cells is important in addition to cellular immunity. However, the role of humoral responses in HSP-based autologous tumor immunotherapy remains unclear. In the present study, we investigated whether leukemia-specific antibodies and antibodymediated cytotoxicity against autologous leukemia cells have a crucial role in a mouse A20 leukemia model by immunizing A20-derived HSP70. Immunization with A20-derived HSP70 induced the production of anti-A20-antibodies and the antibodies recognized HSP70-binding peptides derived from A20. One of those was a major histocompatibility complex ( (1) HSP70 is one of the HSP localized in the cytoplasm and carries peptides to the endoplasmic reticulum, in which glycoprotein (gp)96, another class of HSP, transports the peptides onto the major histocompatibility complex (MHC) class-I molecules.(2-4) In addition, it is known that HSPpeptide complexes are taken up by the antigen-presenting cells (APC) though the cluster of differentiation (CD)91 receptors expressed on APC, and the peptides bound to these HSP are then presented on the MHC class-I or II molecules.(5,6) Because the tumor-derived HSP bind the tumor-specific antigenic peptides, (5)(6)(7) tumor-derived HSP-antigenic peptide complexes are used for vaccination against cancer. As individual HSP are homogeneous, HSP-based cancer vaccine has the advantage of inducing specific immunity against tumor cells of each different haplotype without the need to identify any tumor-specific antigenic peptides. (5,8) In recent years, clinical efficacies of HSP derived from autologous tumor cells have been reported in several cancer patients, including patients suffering renal cell carcinoma, melanoma or chronic myelogenous leukemia, especially in subgroups of patients with relatively few tumors. (9)(10)(11)(12) Although chemotherapies and autologous stem cell transplantations (SCT) have been performed on many patients with leukemia or lymphoma, clinical results have not been satisfactory because of a relapse owing to the presence of minimal residual disease (MRD) and the lack of a graft-versus-leukemia (GVL) effect, both of which can be overcome by allogeneic transplantation. (13)(14)(15) We have established a mouse model with a minimum amount of leukemia cells after syngeneic bone marrow transplantation, and have reported that the immunization of mice with leukemia-cell-derived HSP70 or gp96 induces leukemia-specific immunities by CD8 positive (+) cytotoxic T-cells (CTL) and prolongs the survival of the mice. (16,17) These findings showed the possibility of a clinical application of HSP-based immunotherapy for patients with leukemia, especially for those with minimal residual leukemia cells after SCT.In HSP-based immunotherapy for cancer, cell-mediated immune response by tumor-specific CD8+ CTL is...

We have reported that immunotherapy using leukemia cell-derived heat shock proteins (HSPs) is effective against minimal residual disease (MRD) after syngeneic stem cell transplantation (SCT) in mice. However, leukemia patients after SCT are usually immunocompromised and immunologically tolerant to leukemia cells. We investigated whether the use of dendritic cells (DCs) in combination with HSP70 enhances cytotoxicity against B-cell leukemia cell line A20 in mice after syngeneic SCT. All unimmunized mice died of leukemia early after A20 cell inoculation, whereas mice immunized with HSP70 or HSP70-pulsed DCs survived significantly longer. Although only 60% of the HSP70-immunized mice survived, all mice immunized with HSP70-pulsed DCs survived without MRD. In addition, the cytotoxicities against A20 cells for splenocytes from mice immunized with HSP70-pulsed DCs were significantly higher than those of HSP70-immunized mice, and the cytotoxicities against A20 cells were significantly blocked by anti-CD8 antibody and by major histocompatibility complex class I antibody, but not by anti-CD4 antibody. Moreover, abnormalities were detected in neither the biochemical data nor the histopathologic findings. These findings indicate that the combined use of DCs and leukemia cell-derived HSP70 enhances the antileukemia effect by inducing the specific cytotoxicities of CD8+ cytotoxic T-cells, thereby eradicating MRD effectively and safely, even in an immunocompromised state after syngeneic SCT. This approach may thus be useful for further application of HSP in leukemia patients after autologous SCT.

Mycosis fungoides is a cutaneous T-cell lymphoma, which is clinically divided into three stages: patch, plaque and tumor. Despite a variety of treatments the prognosis is poor in advanced mycosis fungoides. Recently, allogeneic hematopoietic stem cell transplantation has been successfully applied for such cases. We performed reduced-intensity umbilical cord blood transplantation for two advanced mycosis fungoides patients. Case 1 was a 56-year-old man and case 2 was a 30-year-old woman. Tumors of each case were refractory to conventional chemotherapy. Although radiation therapy was considerably effective, tumors relapsed after several months. Reduced-intensity umbilical cord blood transplantation was performed because case 1 had no human leukocyte antigen-identical siblings and the sibling of case 2 did not agree to be the donor. The male patient died of pulmonary failure 23 days after reduced-intensity umbilical cord blood transplantation. The case 2 patient succeeded in reduced-intensity umbilical cord blood transplantation and remained in complete/partial remission for 13 months. However, chemotherapy-resistant tumors relapsed, and allogeneic hematopoietic stem cell transplantation was performed at 17 months. She died of cerebral hemorrhage 23 days after the procedure. Reduced-intensity umbilical cord blood transplantation may be included in the treatments for advanced mycosis fungoides, where graft-versus-lymphoma effect seems to be a significant factor for the success of the treatment.

Most patients with chronic myelogenous leukemia (CML) show a Philadelphia (Ph) chromosome with a characteristic translocation between chromosomes 9 and 22. However, there are variant complex translocations involving other chromosomes in addition to the standard translocation. We describe a case of CML showing a complex and novel chromosomal translocation involving five chromosomes, t(4;12;7;9;22).

The anemia of chronic disease (ACD) is commonly observed in patients with inflammatory disorders, malignancies, and chronic infections. ACD is characterized by low serum iron concentration and elevated serum ferritin concentration. A number of previous findings suggest that impaired mobilization of iron from the reticuloendotherial system (RES), such as macrophages, and an inhibition of iron absorption from intestine contribute to ACD. However, the precise mechanisms for generating the conditions of ACD have long been unclear despite extensive investigations until hepcidin was recently found. Hepcidin was found as one of antimicrobial peptides, but hepcidin is now thought to be a key regulator in iron metabolism. Hepcidin inhibits iron absorption from enterocytes and iron efflux from RES, which have suggested that hepcidin could account for the pathogenesis of ACD. Some cytokines have also been shown to modulate iron metabolism in the condition of inflammation. Interleukin-6 (IL-6), one of inflammatory cytokines, has been reported to induce hepcidin production not only in vitro but also in vivo. However, it is not clear whether other cytokines have such effect or not. We, therefore, investigated the possibility that other inflammatory cytokines might have a regulatory effect on hepcidin expression. Because hepcidin is produced mainly by hepatocytes, we used human hepatoma-derived cell line HuH-7 cells and hepatoblastoma-derived cell line HepG2 cells. Cells are incubated with or without inflammatory cytokines, such as IL-1β, IL-6, tumor necrosis factor α (TNFα), and then total RNA was extracted. Quantitative RT-PCR was then performed, revealing that IL-1β upregulate hepcidin mRNA expression as well as IL-6, although TNFα down regulates hepcidin expression in both cell lines. We next investigated the dose-response effect of IL-1β and IL-6 on hepcidin mRNA expression. Strong induction of hepcidin mRNA by IL-1β was observed when cells were incubated with low concentrations of IL-1β (0.2 ng/ml), although much higher concentrations of IL-6 were needed for hepcidin mRNA upregulation. It was likely that the concentrations of IL-1β that needed for hepcidin upregulation were more physiological than those of IL-6. Since it has been reported that IL-1β induce IL-6 production in hepatocytes, there was a possibility that the effect of IL-1β on hepcidin mRNA expression was not direct but come from IL-6 induced by IL-1β. However, the manners of IL-6 mRNA induction and hepcidin mRNA induction by IL-1β stimulation were quite different when cells were incubated with IL-1β at the concentrations ranging from 0.1 to 10 ng/ml. Moreover, antibody against IL-6 did not inhibit the induction of hepcidin mRNA by IL-1β stimulation. Therefore, it is likely that the effect of IL-1β on hepcidin mRNA expression is independent from that of IL-6. We conclude that inflammatory cytokine IL-1β can induce hepcidin expression and might be a key cytokine in the condition of ACD as well as IL-6, and IL-1β might be more important than IL-6 in physiological situations.

Background: Tumor-derived heat shock proteins (HSPs) can be used for the vaccination against cancer because tumor-derived HSPs bind the tumor specific antigenic peptides and these peptides are carried onto MHC molecules by HSPs. We previously reported that immunotherapy using leukemia cell-derived HSPs against minimal residual leukemia after syngeneic bone marrow transplantation in mice prolonged survival by inducing leukemia-specific CD8 + cytotoxic T lymphocyte (Sato et al. Blood, 2001). It is known that induction of cell-mediated immune response plays a main role of tumor rejection in the HSP-based immunotherapy. However we also showed that CD4+ as well as CD8+ T cell is necessary for survival prolongation of HSP-immunized mice, suggesting that humoral immune response though CD4+ T cell contributes to the eradication of leukemia cells in our mouse model. To investigate the contribution of humoral immune responses in anti-leukemia immunity induced by HSP-based immunotherapy, we tried to detect the anti-leukemia cell antibody and analyzed cytotoxicities by the antibody in the leukemia cell-derived HSP70 immunization mice model. Methods: Balb/c mice and syngeneic A20 B-cell leukemia cell line were used in this study. HSP70 was purified from A20 cells or liver tissue from healthy mice. Three weeks after subcutaneous administration of A20-derived HSP70 (A20-HSP70), liver-derived HSP70 (liver-HSP70) or PBS to the healthy mice, the sera were harvested to perform following experiments. To detect anti-A20 antibodies, mean fluorescence intensity (MFI) of A20 cells with immunized mice sera and FITC-conjugated anti-mouse-IgG was analyzed by flowcytometry. Additionally, the sera were subjected to ELISA using HRP labeled anti-mouse-IgG to detect specific antibody against A20-HSP70. Complement dependent cytotoxicity (CDC) activities were determined by trypan blue uptake of mouse target cells (A20, YAC1: lymphoma, or T27A: myeloid leukemia) after incubation with mice sera and rabbit complement. Results: MIF of A20 with the sera from A20-HSP70 immunized mice was significantly higher than that from PBS or liver-HSP70 immunized mice. In addition, anti-A20-HSP70 IgG level by ELISA was significantly increased in the A20-HSP70 immunized mice. However, anti-A20-HSP70 IgG level in liver-HSP70 mice was not elevated, suggesting that A20-HSP70 immunization induced the specific IgG against not HSP70 itself but peptides bound for A20-HSP70. The sera of mice immunized with A20-HSP70 showed no cytotoxic activity without complement but showed significantly high CDC activity against A20 in vitro. This CDC activity was not observed against YAC-1 or T27A. Conclusions: Leukemia specific CDC by the antibodies against the peptides bound for leukemia cell-derived HSP70 is suggested to be one of the mechanisms of anti-leukemia immunity induced by immunization with leukemia cell-derived HSP70 in mice. This would be an important finding to eradicate leukemia cells effectively in HSP-based immunotherapy for leukemia patients.