Correlation between imatinib pharmacokinetics and clinical response in Japanese patients with chronic-phase chronic myeloid leukemia.

Despite the outstanding results generally obtained with imatinib mesylate (IM) in the treatment of chronic myeloid leukemia (CML), some patients show a poor molecular response. To evaluate the relationship between steady-state trough plasma IM concentration (IM-C(min)) and clinical response in CML patients, we integrated data from six independent Japanese studies. Among 254 CML patients, the mean IM-C(min) was 1,010.5 ng/ml. Importantly, IM-C(min) was significantly higher in patients who achieved a major molecular response (MMR) than in those who did not (P = 0.002). Multivariate analysis showed that an MMR was associated with both age (odds ratio (OR) = 0.97 (0.958-0.995); P = 0.0153) and with IM-C(min) (OR = 1.0008 (1.0003-1.0015); P = 0.0044). Given that patients with IM-C(min) values >1,002 ng/ml had a higher probability of achieving an MMR in our large cohort (P = 0.0120), the data suggest that monitoring of IM levels in plasma may improve the efficacy of IM therapy for CML patients.

Elevation of activated platelet-dependent chemokines and soluble cell adhesion molecules in patients with hematologic malignancies and high levels of beta-D-glucan.

Most invasive fungal infections such as candidemia are frequent in patients with hematologic malignancies. We measured cytokines/chemokines (IL-6, IL-8, monocytic chemoattractant protein 1, RANTES and epithelial neutrophil-activating peptide 78), soluble molecules (sFas, sE-selectin and soluble vascular cell adhesion molecule 1) and platelet activation markers (soluble CD40 ligand, sP-selectin and platelet-derived microparticles) in patients with hematologic malignancies under prophylactic treatment with an antifungal drug (fosfluconazole). We classified patients into 2 groups by the level of beta-D-glucan. The level of C-reactive protein was higher in the high beta-D-glucan group (>5 pg/ml) than in the low beta-D-glucan group. However, there were no differences in the levels of other parameters (peripheral blood cells, glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase, lactate dehydrogenase, blood urea nitrogen and creatinine). Patients in the high beta-D-glucan group exhibited a significant elevation of several chemokines, soluble molecules and platelet activation markers compared with those in the low beta-D-glucan group, but the levels of IL-8, monocytic chemoattractant protein 1 and sFas did not differ significantly. The levels of C-reactive protein and IL-6 increased significantly after 1 or 2 weeks on fosfluconazole in both groups. In contrast, the high beta-D-glucan group exhibited a significant decrease in chemokines, soluble markers and platelet-derived microparticles compared with the low beta-D-glucan group after treatment with fosfluconazole, although the patients in the low beta-D-glucan group exhibited no significant changes. Furthermore, the levels of RANTES, epithelial neutrophil-activating peptide 78, soluble vascular cell adhesion molecule 1 and sE-selectin correlated positively with platelet-derived microparticles in the high beta-D-glucan group. These findings suggest that fungal infection may modulate the vascular events in which some platelet-related chemokines are involved.

Thiol compounds rescue growth inhibition by retinoic acid on HTLV-I (+) T lymphocytes; possible mechanism of retinoic-acid-induced growth inhibition of adult T-cell leukemia cells.

We demonstrated significant growth inhibition by retinoic acid (RA) of HTLV-I (+) T-cell lines (ATL-2 and HUT102), but not HTLV-I (-) T-cell lines (MOLT-4 and Jurkat). We hypothesized that the mechanism of growth inhibition by RA depends on an imbalance in redox potential. To examine the effect of exogenous thiol compounds for the growth of HTLV-I (+) T-cell lines by RA, HTLV-I (+) T-cell lines were cultured with several thiol compounds (thioredoxin, L-cystine, and GSH), following addition of 13-cis RA or ATRA, respectively, in cultured with thiol free medium. Unexpectedly, thiol compounds alone did not restore growth inhibition of HTLV-I (+) T-cell lines. However, when those cells were preincubated with thiol compounds for 24 hours, no growth inhibition by 13-cis RA or ATRA was observed. These results suggest that thiol compounds are associated strongly with sensitivity to RA of HTLV-I (+) T cells, but not of HTLV-I (-) T cells and that thiol compounds serve an important role on HTLV-I (+) T cells.

[Inhibition of proliferation by retinoic acid on adult T cell leukemia cells].

We observed the effects of the retinoic acid (13-cis retinoic acid; 13-cis RA, and all-trans retinoic acid; ATRA) for the cell growth and the expression of CD 25 on peripheral blood mononuclear cells (PBMC) from 17 patients with adult T cell leukemia (ATL). Fourteen had acute type, 1 had chronic type, and 2 had smoldering type of ATL. We divided those patient into 3 groups (hyper-sensitive, sensitive and resistant group) by determined with reduction rate of [3H]-thymidine incorporation obtained before and after treatment with 13 -cis RA or ATRA respectively. Growth inhibition was not observed in normal PBMC by 13 -cis RA or ATRA. However, no down-regulation of CD 25 expression was observed on PBMC in all patients and normal individuals after treatment with 13-cis RA or ATRA. In the aspect of growth inhibition on PBMC in ATL patients, we tried to clarify the mechanism of the phenomenon. In agarose gel electrophoresis, extracted genomic DNA from retinoic acid treated PBMC in hyper-sensitive and sensitive ATL patients showed multimer DNA fragmentation pattern. On the other hand, genomic DNA from PBMC after treatment with retinoic acid in resistant ATL patients and normal individuals showed high molecular DNA pattern without fragmentation. Taken together, it is suggested that retinoic acid could induce growth inhibition of PBMC in some ATL patients resulting in DNA fragmentation, apoptosis. We deeply consider that retinoic acid may be an useful agent for ATL patients in clinical aspect.

Delivery after bone marrow transplantation with total lymphoid irradiation for severe aplastic anemia.

A 28-year old woman delivered a normal child at 85 months after bone marrow transplantation (BMT) for severe aplastic anemia (SAA). BMT was done after conditioning with cyclophosphamide (CY: 50 mg/kg/day for 4 days), anti-lymphocyte globulin (ALG: 50 mg/kg/day for 3 days) and total lymphoid irradiation (TLI: 5 Gy in a single fraction). Ovarian shielding was not done during irradiation. Six months after BMT, she gained normal menstruation spontaneously. Serum gonadotrophin levels were within the normal range, with an LH level of 34.6 mIU/ml (normal: 1-36 mIU/ml) and an FSH level of 13.4 mIU/ml (normal: 1-30 mIU/ml), at 38 months after BMT. She became pregnant 8 years after BMT and delivered a female child, who is now 3 years old and shows normal development.

Elevated levels of soluble ICAM-1 in serum of patients with acute myeloid leukemia undergoing bone marrow transplantation.

Serum soluble ICAM-1 concentrations were measured in 10 patients with or without chronic graft-vs.-host disease (GVHD) after allogeneic bone marrow transplantation. The serum soluble ICAM-1 levels in the patients with chronic GVHD were significantly higher than that in the patients without chronic GVHD. The data indicated that serum soluble ICAM-1 is a useful parameter for predicting chronic GVHD.

Inhibition of growth and induction of apoptosis by all-trans retinoic acid in lymphoid cell lines transfected with the PML/RAR alpha fusion gene.

The interaction of an exogenous PML/RAR alpha fusion gene associated with acute promyelocytic leukaemia, with all-trans retinoic acid (ATRA) was examined in two lymphoid cell lines. L1210 and MOLT-4 cells were transfected with PML/RAR alpha cDNA in the expression vector pGD and stable transformants (L1210PML/RAR alpha and MOLT-4PML/RAR alpha) were selected with G418. ATRA inhibited the growth of these stable transformants, as assessed by [3H]thymidine incorporation, in a dose-dependent manner, but had no effect on the growth of control cells stably transformed with neomycin resistant gene alone. ATRA also induced apoptosis, as assessed by fragmentation of genomic DNA, in L121OPML/RAR alpha and MOLT-4PML/RAR alpha cells but not in control cells. The exogenous PML/RAR alpha fusion gene therefore probably mediates the effects of ATRA on cell growth and apoptosis in these cell lines.

Clonal origin of Philadelphia chromosome negative cells with trisomy 8 appearing during the course of alpha-interferon therapy for Ph positive chronic myelocytic leukemia.

The transient appearance of a Philadelphia chromosome (Ph) negative clone with trisomy 8 was found in the bone marrow cells from a patient with Ph positive CML during the course of alpha-interferon (IFN) therapy. To determine whether this clone was derived from a Ph positive clone or from some other cell lineage, we performed molecular cytogenetic studies on bone marrow cells from the patient by fluorescence in situ hybridization (FISH). No fusion of BCR-ABL could be detected in cells with trisomy 8, clearly indicating that the Ph negative trisomy 8 clone was not derived from the Ph positive CML population.

Change of serum alpha-1 microglobulin and beta-2 microglobulin following allogeneic bone marrow transplantation.

By serially measuring serum levels of alpha-1 microglobulin and beta-2 microglobulin following allogeneic bone marrow transplantation (BMT), we tried to define their relationship to renal dysfunction, acute graft-versus-host disease (GVHD) and infection as complications of the transplantation. The study involved a total of 25 patients with leukemia, myelodysplastic syndrome and aplastic anemia who received BMT in this department; one patient received re-transplantation, thus bringing the total number of transplants to 26. Twenty-four patients received BMT from HLA-identical siblings while two others received BMT from unrelated donors. Alpha-1 microglobulin was within normal limits in all patients before BMT; among various complications such as nephrotoxicity, acute GVHD and infection which took place after transplantation, a raised alpha-1 microglobulin level was found only in nephrotoxicity; however, the increase was not significant compared with the pre-transplantation level. The pre-transplantation beta-2 microglobulin level was higher than normal in some patients; it was significantly increased in all of the above complications compared with the pretransplantation level (1.57 +/- 0.57 mg/l). A significant correlation was found between the serum creatinine level and the beta-2 microglobulin level (r = 0.849) in patients with renal dysfunction. In some patients, however, the beta-2 microglobulin level increased earlier than the serum creatinine level, and this finding was considered useful for the early diagnosis of renal dysfunction following allogeneic BMT.

[Unrelated match bone marrow transplantation for severe aplastic anemia].

The results of unrelated bone marrow transplantation (BMT) is poor because of the rejection of bone marrow graft and graft versus host disease (GVHD). However, the rate of rejection has been reported to be decreased by intensive immuno-suppressive preconditioning regimens combined with total body irradiation (TBI). We report a case of an 18-year-old male with severe aplastic anemia who received a matched BMT from an unrelated donor. The pre-conditioning regimen included cyclophosphamide (50mg/kg) for 4 days, total lymphoid irradiation (TLI: 6Gy) and TBI (5Gy). GVHD (grade 1), hemorrhage cystitis and varicella occurred after BMT but were cured. His performance status is now 100% on the Karnofsky score at 10 months after BMT.

[Granulopoietic inhibitor].

The differentiation and proliferation of granulocytes are specifically supported by G-CSF. On the other hand, little is known about the negative modulation of granulopoiesis. We had found that the sera obtained from patients during the recovery phase of hematopoiesis after chemotherapy inhibited the formation of CFU-G, suggesting the modulation of granulopoiesis by negative regulator(s). As the inhibitory effect was specific for CFU-G, the inhibitor in this inhibitory sera was suggested the soluble G-CSF receptor. Several G-CSF receptor fragments were detected in the concentrated fractionated-urine by the immunoblot analysis using the specific antibody against the cytokine receptor homologous (CRH) domain of G-CSF receptor. G-CSF binding protein was successfully obtained from normal human plasma by G-CSF affinity column. Major component was the 90Kd of protein which was thought to be the extracellular domain of G-CSF receptor by the immunoblot analysis. About 90Kd- and 30Kd- bands of the binding-complex formation between G-CSF monomer and the extracellular domain or the fragment of G-CSF receptor were shown in the immunoblot analysis using anti-G-CSF Ab. CFU-G formation was suppressed by the solution of soluble G-CSF receptor fragments, suggesting the competition of G-CSF which bind to granulocyte precursor cells.

Inhibition of CFU-G formation by human serum during granulopoiesis after chemotherapy: purification of CFU-G-inhibitory factor and its activities.

We found that the sera obtained from patients with acute leukemia (AL) in complete remission or malignant lymphoma (ML) during the recovery phase after chemotherapy completely inhibited the GCT-CM-stimulated growth of allogenic and autologous bone marrow colonies in vitro. We investigated 5 AL patients and 6 ML patients from whom blood samples were taken either every day or every 6 h during the recovery phase after chemotherapy. Colony-inhibitory sera, were detected in all patients, more frequently when the peripheral leukocyte count recovered to about 2,500 x 10(6)/l, but then transiently and at intervals. The colony-inhibitory factor purified from the inhibitory sera inhibited the growth of G-CSF-responsive colonies in a dose-dependent manner, but no effect on the growth of GM-CSF-responsive colonies, CFU-E, or BFU-E, was observed. These results suggest that this factor may play a role in regulating granulopoiesis by inhibiting the differentiation of G-CSF-responsive precursor cells.

[Effect of recombinant human granulocyte colony-stimulating factor on white blood cell count and neutrophil functions in patients with malignant lymphoma].

We studied the in vivo effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on white blood cell (WBC) count and neutrophil functions in nine patients with malignant lymphoma. The WBC count and absolute neutrophil count were rapidly increased without a nadir phase after chemotherapy. Neutrophil alkaline phosphatase (NAP) scores also markedly increased following chemotherapy in all patients. Phagocytosis of India ink and nitroblue tetrazolium (NBT) reduction were revealed tend to be increased, but not exceeded significantly to normal range. RhG-CSF repaired neutrophil function in patients with decreasing that. Thus, rhG-CSF may be useful for prevention and treatment of infection after chemotherapy.