Haemotoxicity of busulphan, doxorubicin, cisplatin and cyclophosphamide in the female BALB/c mouse using a brief regimen of drug administration.
Gemma Molyneux, Michael Andrews, William Sones, Malcolm York, Anne Barnett, Edel Quirk, Wing Yeung, John Turton
Department of Pharmacology, Centre for Toxicology, The School of Pharmacy, University of London, 29/39 Brunswick Square, London, UK. email@example.com
Many anticancer drugs are myelotoxic and cause bone marrow depression; however, generally, the marrow/blood returns to normal after treatment. Nevertheless, after the administration of some anti-neoplastic agents (e.g. busulphan, BU) under conditions as yet undefined, the marrow may begin a return towards normal, but normality may not be achieved, and late-stage/residual marrow injury may be evident. The present studies were conducted to develop a short-term mouse model (a 'screen') to identify late-stage/residual marrow injury using a brief regimen of drug administration. Female BALB/c mice were treated with BU, doxorubicin (DOX), cisplatin (CISPLAT) or cyclophosphamide (CYCLOPHOS) on days 1, 3 and 5. In 'preliminary studies', a maximum tolerated dose (MTD) for each drug was determined for use in 'main studies'. In main studies, mice were treated with vehicle (control), low and high (the MTD) dose levels of each agent. Necropsies were performed, and blood parameters and femoral/humeral nucleated marrow cell counts (FNCC/HNCC) were assessed on six occasions (from days 1 to 60/61 post-dosing). Late-stage/residual changes were apparent in BU-treated mice at day 61 post-dosing: RBC, Hb and haematocrit were reduced, mean cell volume/mean cell haemoglobin were increased and platelet and FNCC counts were decreased. Mice given DOX, CISPLAT and CYCLOPHOS, in general, showed no clear late-stage/residual effects (day 60/61). It was concluded that a brief regimen of drug administration, at an MTD, with assessment at day 60/61 post-dosing was a suitable short-term method/screen in the mouse for detecting late-stage/residual marrow injury for BU, a drug shown to exhibit these effects in man.
Immunol Invest. 2006 ;35 (2):209-25 16698678
Cord blood-derived plasma factor (CBPF) potentiates the low cytokinetic and immunokinetic profile of bone marrow cells in pesticide victims suffering from Acquired Aplastic Anaemia (AAA): an in vitro correlate.
Department of Haematology, Immunology and Immuno-Haematology Laboratory, School of Tropical Medicine, C.R. Avenue, Kolkata, India.
Cases of Acquired Aplastic Anaemia (AAA) in patients with a long history of pesticide exposure from agricultural fields have been investigated in our laboratory using an immunological approach. These patients showed moderate to severe degrees of bone marrow aplasia as a result of 9-12 years protracted exposure to pesticides which were mainly comprised of organophosphorous and organochloride compounds. The bone marrow aspirate culture was found to be severely deficient both in terms of differentiation and proliferation, and cell mediated immune function (CMI). We attempted ex vivo manipulation of the bone marrow population of patients in two different protocols: in one, stem cell factor (SCF), interleukin-3 (IL-3), and granulocyte-colony stimulating factor (G-CSF) were administered and, in the second set, cord blood-derived plasma factors (CBPF) were supplemented to evaluate the effects, if any. Simultaneously, two control groups including one for healthy normal control (N) and the second, for non-pesticide induced aplastic anaemia group of patients (NPAA) was also investigated for all the above parameters. Active colony formation and improved cellular immune activity (CMI) was observed more frequently in the CBPF treated group rather than that in the cytokine treated group. Surprisingly, administration of cytokines in the first set and CBPF in the second set triggered CD34 (+) cell generation as revealed through flow cytometric analysis (FACS). The effect was more pronounced in the second set. Investigations carried out with NPAA showed relatively insignificant effects with both cytokine and CBPF set up. The investigations indicated that AAA as induced by pesticides could be therapeutically manipulated by exogenous cytokines and growth factors and, more efficiently, by CBPF by way of immunopotentiation through microenvironmental supplementation.
Gemma Molyneux, Frances M Gibson, Edward C Gordon-Smith, Andrew M Pilling, Kai Chiu Liu, Sian Rizzo, Susan Sulsh, John A Turton
Department of Haematology, St George's Hospital Medical School, London, UK.
Mitomycin (MMC), like many antineoplastic drugs, induces a predictable, dose-related, bone marrow depression in man and laboratory animals; this change is generally reversible. However, there is evidence that MMC may also cause a late-stage or residual bone marrow injury. The present study in female CD-1 mice investigated the haematological and bone marrow changes induced by MMC in a repeat dose study lasting 50 days. Control and MMC-treated mice were dosed intraperitoneally on eight occasions over 18 days with vehicle, or MMC at 2.5 mg/kg, autopsied (n = 6-12) at 1, 7, 14, 28, 42 and 50 days after the final dose and haematological changes investigated. Femoral nucleated bone marrow cell counts and levels of apoptosis were also evaluated and clonogenic assays carried out; serum levels of FLT3 ligand (FL) were assessed. At day 1 post-dosing, MMC induced significant reductions in RBC, Hb and haematocrit (HCT) values, and there were decreases in reticulocyte, platelet, and femoral nucleated cell counts (FNCC); neutrophil, lymphocyte and monocyte values were also significantly reduced. On days 7 and 14 post-dosing, all haematological parameters showed evidence of a return towards normal values, but at these times, and at day 28, values for RBC and FNCC remained significantly reduced in comparison with controls. At days 42 and 50 post-dosing, many haematological parameters in MMC-treated mice had returned to control levels; however, there remained evidence of late-stage effects on RBC, Hb and HCT values, and FNCC also continued to be significantly decreased. Results for granulocyte-macrophage colony-forming units and erythroid colonies showed a profound decrease immediately post-dosing, but a return to normal values was evident at day 50. Serum FL concentrations demonstrated very significant increases in the immediate post-dosing period, but a return to normal was seen at day 50 post-dosing; a relatively similar pattern was seen in the number of apoptotic femoral marrow nucleated cells. The histopathological examination of kidney tissues from MMC animals at day 42 and 50 post-dosing showed evidence of hydronephrosis with cortical glomerular/tubular atrophy and degeneration. It is therefore concluded that MMC administered on eight occasions over 18 days to female CD-1 mice at 2.5 mg/kg induced profound changes in haematological and bone marrow parameters in the immediate post-dosing period with a return to normal levels at day 50 post-dosing; however, there was evidence of mild but significant late-stage/residual effects on RBC and FNCC, and on cells of the erythroid lineage in the bone marrow.
Marked decreases of total and immature reticulocytes in myelodysplastic syndrome among patients with pancytopenia.
Central Laboratory for Clinical Investigation, Osaka University Hospital, Suita, Japan. firstname.lastname@example.org
Frances M Gibson, C Michael Andrews, Paraskevi Diamanti, Sian Rizzo, George Macharia, Edward C Gordon-Smith, Thomas Williams, John Turton
Department of Haematology, St George's Hospital Medical School, London SW17 0RE, UK.
Aplastic anaemia (AA) is characterized by hypocellular marrow, pancytopenia, and risk of severe anaemia, haemorrhage and infection. AA is often idiopathic, but frequently occurs after exposure to drugs/chemicals. However, the pathogenesis of AA is not clearly understood, and there are no convenient animal models of drug-induced AA. We have evaluated regimens of busulphan (BU) administration in the mouse to produce a model of chronic bone marrow aplasia showing features of human AA. Mice were given 8 doses of BU at 0, 5.25 and 10.50 mg/kg over 23 days; marrow and blood samples were examined at 1, 19, 49, 91 and 112 days after dosing. At day 1 post dosing, in mice treated at 10.50 mg/kg, nucleated marrow cells, CFU-GM and Erythroid-CFU were reduced. Similarly, peripheral blood erythrocytes, leucocytes, platelets and reticulocytes were reduced. At day 19 and 49 post dosing, there was a trend for parameters to return towards normal. However, at day 91 and 112 post dosing, values remained significantly depressed, with a stabilized chronic bone marrow aplasia. At day 91 and 112 post dosing, marrow cell counts, CFU-GM and Erythroid-CFU were decreased; marrow nucleated cell apoptosis and c-kit+ cell apoptosis were increased; peripheral blood erythrocyte, leucocyte, and platelet counts were reduced. We conclude that this is a model of chronic bone marrow aplasia which has many interesting features of AA. The model is convenient to use and has potential in several areas, particularly for investigations on mechanisms of AA pathogenesis in man.
An assessment of chloramphenicol and thiamphenicol in the induction of aplastic anaemia in the BALB/c mouse.
Centre for Toxicology, Department of Pharmacology, The School of Pharmacy, University of London, UK. email@example.com
The potential of the antibiotics chloramphenicol succinate (CAPS) and thiamphenicol (TAP) to induce aplastic anaemia in the female BALB/c mouse was investigated. CAPS was administered at 2000 mg/kg, and TAP at 850 mg/kg, daily by gavage, for 17 days. At 1, 13, 22, 41, 98 and 179 days after the final dose of each antibiotic, mice (n = 4 or 5) were sampled for haematological examination and haematopoietic stem cell assays. Both CAPS and TAP induced significant reductions in red blood cell count, haematocrit and haemoglobin values at day 1 post dosing; counts of colony-forming units-erythroid and colony-forming units-granulocyte-macrophage, were similarly significantly decreased at this time. All these reduced parameters returned towards normal at days 13 and 22. At days 41, 98 and 179, results for all haematological values and stem cell assays in both CAPS- and TAP-treated mice compared with the controls; there was no evidence of a reduction in peripheral blood values or bone marrow parameters at the later sampling points, as would be expected in a developing or overt bone marrow aplasia. We therefore consider that the administration of CAPS and TAP, which have been associated with the development of aplastic anaemia in man, induce a reversible anaemia, but not a chronic bone marrow aplasia, when given at haemotoxic dose levels for 17 days in the BALB/c mouse.
Pathophysiological significance of simultaneous measurement of reticulated platelets, large platelets and serum thrombopoietin in non-neoplastic thrombocytopenic disorders.
Department of Clinical and Laboratory Medicine, Osaka City University Medical School, Osaka, Japan.
An automated reticulocyte counter using flowcytometric analysis, the R-3000 (Sysmex Inc. Kobe, Japan), has recently been modified to determine reticulated platelets (RPs) and large platelets (LPs). We measured frequencies of RPs, LPs in total platelet count and serum thrombopoietin concentration comprehensively in non-neoplastic thrombocytopenic patients with immune thrombocytopenic purpura (ITP, n = 23), aplastic anemia (AA, n = 21), liver cirrhosis (LC, n= 17), and hematologically normal subjects (control, n = 151). ITP was characterized as high frequencies of both RP and LP, AA as high RP frequency and elevated thrombopoietin concentration, and LC as no difference compared with control. Interestingly, the frequency of RP appeared to depend on total platelet count rather than the cause of thrombocytopenia, while the frequency of LP appeared to depend much less on total platelet count. Furthermore, significant positive correlations were observed between frequencies of RP and LP in control, ITP and LC subjects, in whom bone marrow stem cells are intrinsically normal. However, there was no such correlation in AA patients with stem cell deficiency, suggesting that this correlation might be a useful new parameter for detecting qualitatively abnormal platelets. Measurement of RP and LP is thus useful for elucidating the pathophysiology of thrombocytopenic disorders.
Use of human lymphoblastoid cells to detect the toxic effect of chloramphenicol and metabolites possibly involved in aplastic anemia in man.
Laboratory of Molecular Genetics, UPR 42, CNRS, Villejuif, France.
Some Chloramphenicol (CAP) metabolites are suspected to be involved in the etiology of bone marrow aplasia in man. The objective of the present study was to investigate the cytotoxicity as well as the genotoxicity of CAP and six of its metabolites on human bone marrow cells (RiBM cells) and to compare these results with those obtained on human peripheral blood lymphocytes in order to estimate the relative sensitivity of the two types of cells. Three CAP metabolites NO-CAP, DH-CAP and NPAP inhibited 3H thymidine incorporation in RiBM cells at concentrations ranging from 2.10(-5) M to 2.10(-4) M. NO-CAP appeared as the most potent cytotoxic compound. CAP itself and NAPD presented some toxic effect at high concentration (1-2.10(-3) M). CAPG and HAP did not present any cytotoxic effect. By comparison, the response of human lymphocytes to CAP and its metabolites showed a similar pattern but DH-CAP was the most inhibitory compound. Concerning the genotoxic potential, NO-CAP and DH-CAP induced DNA single strand breaks in RiBM cells at concentrations of 1 and 2.10(-4) M with a dose response relationship. CAP and other metabolites were completely devoid of genotoxicity up to 4.10(-3) M. The results clearly showed that RiBM cells were much less susceptible to the genotoxic effect of CAP metabolites than human lymphocytes.
Department of Laboratory Medicine, School of Medicine, Keio University, Tokyo, Japan.
AIMS--To determine the maturity of reticulocytes in patients with anaemia as a result of various haematological disorders including those with qualitative abnormalities such as ineffective erythropoiesis or dyserythropoiesis. METHODS--The number of mature reticulocytes was measured with flow cytometry in venous blood samples from 122 patients with haematological disorders and 100 healthy controls. Reticulocytes were classified into three categories by the fluorescence intensity of auramin O staining: low fluorescence ratio (LFR), medium fluorescence ratio (MFR), and high fluorescence ratio (HFR). Immature reticulocytes were determined as the aggregate of MFR and HFR (%). RESULTS--The mean (2SD) number of immature reticulocytes in 100 normal subjects was 9.0 (7.0)%. Significantly high mean values of immature reticulocytes with a normal or reduced reticulocyte count were shown in 90 patients with dyserythropoietic or ineffective erythropoietic conditions, such as acute myeloid leukaemia (AML)(n = 37), myelodysplastic syndrome (MDS)(n = 35), aplastic anaemia (AA)(n = 8), or megaloblastic anaemia (MA),(n = 6). Reticulocyte ratios returned to normal after successful treatment of patients with AML (n = 10) and MA (n = 3). However, high percentages of immature reticulocytes with increased reticulocyte counts were consistently observed in patients with enhanced erythropoiesis such as those with acquired autoimmune haemolytic anaemias (AIHA)(n = 4) or acute blood loss (ABL)(n = 4). Reticulocyte maturity was within the normal range in patients with reduced erythropoiesis such as occurs in chronic renal failure (CRF)(n = 11), or in iron deficiency anaemia (IDA)(n = 13). CONCLUSIONS--The evaluation of reticulocyte maturity with total reticulocyte count seems to be clinically useful for estimating the qualitative impairment of erythropoiesis, and so could help differentiate haematological disorders.
Other papers by authors:
Allogeneic and syngeneic marrow transplantation for myelodysplastic syndrome in patients 55 to 66 years of age.
H J Deeg, H M Shulman, J E Anderson, E M Bryant, T A Gooley, J T Slattery, C Anasetti, A Fefer, R Storb, F R Appelbaum
Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA. firstname.lastname@example.org
We carried out bone marrow transplantation (BMT) in 50 patients with myelodysplastic syndrome (MDS) who were 55.3 to 66.2 years of age (median, 58.8 years). According to the criteria of the French-American-British (FAB) classification, 13 patients had refractory anemia (RA), 19 had RA with excess blasts (RAEB), 16 had RAEB in transformation or acute myelogenous leukemia (RAEB-T/AML), and 2 had chronic myelomonocytic leukemia (CMML). According to the recently established International Prognostic Scoring System (IPSS), available for 45 patients, 2 patients were considered low risk; 14, intermediate 1 risk; 19, intermediate 2 risk; and 10, high risk. Conditioning regimens were cyclophosphamide (CY)(120 mg/kg of body weight) plus 12-Gy fractionated total-body irradiation (FTBI)(n = 15), CY plus FTBI with lung and liver shielding (n = 4), busulfan (7 mg/kg) plus FTBI (n = 4), or busulfan (16 mg/kg) plus CY (n = 27). The busulfan-plus-CY group included 16 patients in whom busulfan was targeted to plasma levels of 600 to 900 ng/mL. In these 16 patients, steady-state levels of busulfan actually achieved were 714 to 961 ng/mL (mean +/- SD, 845 +/- 64 ng/mL; median, 838 ng/mL). The donors were HLA-identical siblings for 34 patients, HLA-nonidentical family members for 4, identical twins for 4, and unrelated volunteers for 6. All 46 patients surviving > 21 days had engraftment, and 22 patients (44%) are surviving 9 to 80 months after BMT. Specifically, among 13 patients with RA, 1 had relapse (cumulative incidence [CI] at 3 years, 8%) and 8 are surviving, for a Kaplan-Meier (KM) estimate of survival at 3 years of 59%(disease-free survival [DSF], 53%). Among 19 patients with RAEB, 3 had relapse (CI at 3 years, 16%), and 8 are surviving disease free (KM estimate at 3 years, 46%). Among 18 patients with RAEB-T/AML or CMML, 6 had relapse (CI at 3 years, 28%), and the KM estimate of DSF at 3 years is 33%. Relapse-free survival had an inverse correlation with cytogenetic risk classification and with the risk score according to the IPSS. Survival in all FAB categories was highest among patients enrolled in a protocol in which busulfan plasma levels were targeted to 600 to 900 ng/mL. These data indicate that BMT can be carried out successfully in patients with MDS who are older than 55 years of age.(Blood. 2000;95:1188-1194)
Interleukin-2 after autologous stem cell transplantation for hematologic malignancy: a phase I/II study.
N Robinson, M C Benyunes, J A Thompson, A York, S Petersdorf, O Press, C Lindgren, T Chauncey, C D Buckner, W I Bensinger, F R Appelbaum, A Fefer
Department of Medicine, University of Washington, the Fred Hutchinson Cancer Research Center, Seattle 98195, USA.
The success of autologous stem cell transplantation (ASCT) for hematologic malignancy is limited largely by a high relapse rate. It is postulated that IL-2 administered after ASCT may eliminate minimal residual disease and thereby reduce relapses. A phase I/II study was performed to identify a regimen of IL-2 (Chiron) that could be given early after ASCT in phase III trials. In the phase I study, beginning a median of 46 days after ASCT for hematologic malignancy, cohorts of three to four patients received escalating doses of 'induction' IL-2 of 9, 10, or 12 x 10(6) IU/m2/day for 4 or 5 days by continuous i.v. infusion (CIV), followed by a 4-day rest period, and then 1.6 x 10(6) IU/m2/day of maintenance IL-2 by CIV for 10 days. The maximum tolerated dose (MTD) of induction IL-2 was 9 x 10(6) IU/m2/day x 4. In the phase II study, 52 patients received the MTD. Eighty percent of patients completed induction IL-2. Most patients exhibited some degree of capillary leak. One patient died of CMV pneumonia and one died of ARDS. Maintenance IL-2 was well tolerated. In the phase I/II study, 16 of 31 patients with non-Hodgkin lymphoma (NHL), 3/8 with Hodgkin disease (HD), 4/17 with AML, and 4/5 with ALL remain in CR. Two of six multiple myeloma (MM) patients remain in PR. Although the regimen of IL-2 identified had significant side-effects in some patients, it was well tolerated in the majority of patients. Phase III prospectively randomized clinical trials are in progress to determine if this IL-2 regimen will decrease the relapse rate after ASCT for AML and NHL.
W I Bensinger, T Demirer, C D Buckner, F R Appelbaum, R Storb, K Lilleby, P Weiden, A Z Bluming, A Fefer
Fred Hutchinson Cancer Research Center, University of Washington, School of Medicine, Seattle, USA.
Eleven patients with advanced multiple myeloma (MM) received syngeneic marrow (n = 10) or peripheral blood stem cell (n = 1) transplants following cyclophosphamide (CY) and total body irradiation (TBI)(n = 8), busulfan (Bu) and CY (n = 1), Bu, CY and TBI (n = 1) or Bu, melphalan and thiotepa (n = 1). At the time of transplant one patient had stage II and 10 patients had stage III disease. Four patients had refractory disease, two had chemotherapy sensitive disease and five had progressed after an initial response to chemotherapy. The median time from diagnosis to transplant was 353 days (range 176-6118). After transplant, the median time to achieve granulocytes of 0.5 x 10(9)/l and platelets of 20 x 10(9)/l was 12 days (range 9-20) and 12 days (9-27), respectively. One patient died of interstitial pneumonia syndrome on day 32 and one died of veno-occlusive disease of the liver on day 44 post-transplant, and these were unevaluable for response. Five of nine evaluable patients achieved a complete response (CR), three a partial response, and one patient had no response. Three patients who did not achieve CR died of progressive disease 106, 142 and 321 days post-transplant. Of five patients who achieved a CR, three relapsed on days 539, 737 and 1706 and died on days 1759, 1596 and 1736, respectively; one patient died of myelodysplastic syndrome on day 1407 without evidence of MM and one patient is alive and disease-free 3297 days after transplant. One of the two long-term survivors has a persistent monoclonal protein in the blood 15 years post-transplant. These data show that high-dose therapy and infusion of normal syngeneic marrow cells can cure a small fraction of patients with MM. However, the majority of patients did not achieve durable CR, demonstrating the need for improved transplant conditioning regimens, earlier transplant or additional post-transplant treatment strategies when syngeneic transplants are performed.
High-dose fractionated total-body irradiation, etoposide and cyclophosphamide for treatment of malignant lymphoma: comparison of autologous bone marrow and peripheral blood stem cells.
M W Brunvand, W I Bensinger, E Soll, C H Weaver, S D Rowley, F R Appelbaum, K Lilleby, R A Clift, T A Gooley, O W Press, A Fefer, R Storb, J E Sanders, P L Martin, T Chauncey, R T Maziarz, N Zuckerman, P Montgomery, R Dorn, P L Weiden, T Demirer, L A Holmberg, K Schiffman, P A McSweeney, C D Buckner
Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
Consecutive patients with non-Hodgkin's lymphoma (NHL, n = 133) or Hodgkin's disease (HD, n = 20) were treated with 12.0 Gy of fractionated total body irradiation, etoposide 60 mg/kg, and CY 100 mg/kg followed by infusion of autologous hematopoietic stem cells. Seventy-nine patients received purged (n = 62) or unpurged BM (n = 17), and 74 received unpurged PBSCs alone (n = 56) or with BM (n = 18). The median day for achieving a sustained granulocyte count of 0.5 x 10(9)/I was 14 range (7-66) for BM recipients and 10 (7-30) for PBSC +/- BM recipients (P = 0.03). A platelet count of 20 x 10(9)/I was achieved at a median of day 24 (6-145) in BM recipients and day 11 (range, 7-56) in PBSC +/- BM recipients (P = 0.007). The median number of platelet units transfused was 86 (0-1432) for BM recipients and 30 (6-786) for PBSC +/- BM recipients (P = 0.001). The median number of hospital days was 36 (10-88) for BM recipients and 27 (14-76) for PBSC +/- BM recipients (P = 0.0001). The unadjusted Kaplan-Meier (KM) estimates of survival, event-free survival (EFS) and relapse at 2 years were 0.57, 0.45 and 0.43 for patients receiving BM and 0.55, 0.36 and 0.59 for patients receiving PBSC +/- BM. After adjusting for confounding variables, the estimated relative risk (RR) of death from any cause was 0.92 (P = 0.75), of relapse was 1.25 (P = 0.39), of non-relapse mortality was 0.71 (P = 0.42) and of mortality and/or relapse was 1.17 (P = 0.48) for patients receiving PBSC +/- BM as compared to BM. For 46 patients with NHL receiving unpurged PBSC alone, the unadjusted KM estimate of relapse was 0.61 compared with 0.48 for 52 comparable patients receiving purged BM, while the RR for relapse for patients receiving unpurged PBSCs was 1.37 (P = 0.33) after adjusting for other significant covariates. These data confirm previous observations that patients who receive PBSC +/- BM have faster engraftment, fewer transfusions and shorter hospital stays than patients who receive only BM. There were no statistically significant differences between the two groups in survival, relapse, death from causes other than relapse and event-free survival.
Autologous transplantation with peripheral blood stem cells collected after granulocyte colony-stimulating factor in patients with acute myelogenous leukemia.
T Demirer, F B Petersen, W I Bensinger, F R Appelbaum, A Fefer, S Rowley, J Sanders, T Chauncey, R Storb, K Lilleby, C D Buckner
Fred Hutchinson Cancer Research Center. Seattle, WA 98104, USA.
The use of peripheral blood stem cells (PBSC) with or without bone marrow (BM) in patients with acute myelogenous leukemia (AML) undergoing autologous transplantation in untreated first relapse (Rel1) or in second remission (CR2) was evaluated in a phase II study. Twenty-three patients with AML in untreated Rel1 (n = 8) and CR2 (n = 15) underwent autologous transplant using PBSC with (n = 19) or without (n = 4) BM. Six patients received busulfan (BU) and cyclophosphamide (CY) and 17 received BU, CY and total body irradiation prior to transplant. The median number of CD34+ cells infused was 4.81 x 10(6)/kg (range 0.04-15). Fifteen of 23 patients received post-transplant interleukin-2 (IL-2) at a median of 43 days (range 11-93) in an attempt to decrease relapses. The median day of recovery of granulocytes to 0.5 x 10(9)/I was 12 (range 8-27) and platelets to 20 x 10(9)/I was 15 (range 8-103). Patients received a median of 4 units (range 0-20) of red blood cells and 29 units (range 4-252) of platelets. The probability of 100 day non-relapse mortality was 0.14. The probabilities of survival and relapse at 2 years were 0.24 and 0.65, respectively. The probabilities of relapse in patients receiving (n = 15) and not receiving (n = 8) interleukin-2 (IL-2) were 0.59 and 0.74, respectively (P = 0.1). Overall, seven of 23 (30%) patients are alive and continuously disease-free at a median of 483 days (range 113-835) post-transplant. These data demonstrate that the infusion of PBSC collected after rhG-CSF corrected engraftment problems previously observed with autologous BM transplants in patients with AML but was associated with a high relapse rate.
Busulfan, cyclophosphamide and fractionated total body irradiation for autologous or syngeneic marrow transplantation for acute and chronic myelogenous leukemia: phase I dose escalation of busulfan based on targeted plasma levels.
T Demirer, C D Buckner, F R Appelbaum, W I Bensinger, J Sanders, K Lambert, R Clift, A Fefer, R Storb, J T Slattery
Fred Hutchinson Cancer Research Center, Seattle, WA 98104, USA.
In a previous phase I study, it was concluded that tolerable doses of busulfan (BU), cyclophosphamide (CY) and total body irradiation (TBI) were 8 mg/kg, 60 mg/kg and 12.0 Gy, respectively, for autologous marrow transplant recipients. In an attempt to decrease the variability of BU steady-state concentration (Css) following oral dosing, a BU dose escalation study based on targeted plasma levels was performed in patients receiving autologous transplants for AML or syngeneic transplants for CML. In this study, the BU dose was adjusted up or down based on observed plasma concentration. All patients received a fixed dose of CY 60 mg/kg and TBI of 12 Gy. The first dose level evaluated was 8.6 mg/kg with a target BU Css of 511 ng/ml. Eight patients were entered at this level and the median BU Css achieved was 441 (range 253-566). One of eight patients developed grade 3-4 regimen-related toxicities (RRT). The oral dose of BU for dose level II was 10.6 mg/kg with a target Css of 632 ng/ml. Six patients were entered at this level and median BU Css achieved was 642 (range 566-674). One of six patients developed grade 3-4 RRT. The oral dose for dose level III was 12.6 mg/kg with a target BU Css of 754 ng/ml. Five patients with AML were entered at this dose level and the median plasma BU Css was 733 ng/ml (682-900). Two of five (40%) patients at dose level III developed grade 3-4 RRT which was considered excessive making dose level II the MTD. This study showed that targeted BU Css can reliably be achieved with a bias of -5.23% and mean absolute error of 11.3%. Overall, targeting made a -32.5% to 158.3% change in plasma BU Css as compared to expected BU Css based on first dose pharmacokinetics if targeting were not performed in this study. Thus, targeting avoided much of the variability in BU Css seen in other studies and appears to have allowed for an increase in oral dosing from 8 mg/kg to 10.6 mg/kg. Despite achieving higher and more uniform BU Css, there was no apparent effect on relapse or survival, although the number of patients evaluated was small.
High-dose therapy followed by autologous hematopoietic stem-cell infusion for patients with multiple myeloma.
W I Bensinger, S D Rowley, T Demirer, K Lilleby, K Schiffman, R A Clift, F R Appelbaum, A Fefer, T Barnett, R Storb, T Chauncey, R T Maziarz, J Klarnet, P McSweeney, L Holmberg, D G Maloney, C H Weaver, C D Buckner
Fred Hutchinson Cancer Research Center, Seattle, WA 98104, USA.
PURPOSE: To evaluate the outcome of patients with multiple myeloma (MM) who received high-dose therapy followed by autologous bone marrow (BM) or peripheral-blood stem-cell (PBSC) infusion. PATIENTS AND METHODS: Sixty-three consecutive patients with MM received autologous BM (n = 13) or PBSC with or without BM (n = 50) following regimens that contained busulfan (Bu) and cyclophosphamide (Cy)(n = 18), modified total-body irradiation (TBI) followed by Bu and Cy (n = 36), or Bu, melphalan, and thiotepa (n = 9). Two thirds of the patients had resistant disease and 69% had received more than 6 months of previous chemotherapy. RESULTS AND CONCLUSION: Recovery of peripheral-blood cell counts was more rapid in patients who received PBSC with or without BM than in patients who received BM alone. Sixteen of 63 patients (25%) died of complications of treatment within 100 days. Nineteen (40%) of 48 assessable patients achieved a complete response (CR), 23 (48%) had a partial response (PR), and six (12%) had no response. The probabilities of survival and survival without relapse or progression for all 63 patients at 3.0 years were .43 and .21, respectively. The probability of relapse or progression at 3 years was .69, and 17 patients (27%) have died of progressive MM. The probabilities of survival and relapse-free survival at 3 years for the 19 patients who achieved a CR were .42 and .17, respectively. In the multivariate analysis, beta2-microglobulin levels more than 2.5 micrograms/mL, more than two regimens of prior therapy and eight cycles of treatment, time to transplant longer than 3 years from diagnosis, and prior radiation were associated with adverse outcomes. Additional strategies, such as intervention earlier in the disease course, improved treatment regimens, sequential high-dose treatments, and posttransplant therapies may improve outcome of selected patients with MM.
Phase I trial of interleukin-2 after unmodified HLA-matched sibling bone marrow transplantation for children with acute leukemia.
Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
Allogeneic bone marrow transplantation (BMT) for advanced acute leukemia is associated with a high risk of relapse. It is postulated that interleukin-2 (IL-2) administered after BMT might induce or amplify a graft-versus-leukemia effect and thereby reduce the relapse rate. To identify an IL-2 regimen for testing this hypothesis, a phase I trial of IL-2 (Roche) was performed in children in complete remission (CR) without active graft-versus-host disease (GVHD) off immunosuppressive agents after unmodified allogeneic matched-sibling BMT for acute leukemia beyond first remission. Beginning a median of 68 days after BMT, 17 patients received escalating doses of induction IL-2 (0.9, 3.0, or 6.0 x 10(6) IU/m2/d representing levels I, II, and III) for 5 days by continuous intravenous infusion (CIV). After 6 days of rest, they received maintenance IL-2 (0.9 x 10(6) IU/m2/d) for 10 days by CIV infusion. Levels I and II were well-tolerated, but, of 6 patients at level III, 1 developed pulmonary infiltrates, 1 developed hypotension (both resolved), and 1 died of bacterial sepsis and acute respiratory distress syndrome. Grade II acute GVHD developed in 1 patient at level I and 1 at level III. The maximum tolerated dose of induction IL-2 was level II. IL-2 induced lymphocytosis, with an increase in CD56+ and CD8+ cells. Ten patients remain in CR at 5+ to 67+ months. Thus, a regimen of IL-2 has been identified that did not induce a high incidence of acute GVHD when administered to children after unmodified allogeneic BMT. Its clinical activity will be assessed in a phase II trial.
Autologous marrow transplantation for patients with acute myeloid leukemia in untreated first relapse or in second complete remission.
F B Petersen, M H Lynch, R A Clift, F R Appelbaum, J E Sanders, W I Bensinger, M C Benyunes, K Doney, A Fefer, P Martin
Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
PURPOSE This study compares outcomes of autologous bone marrow transplantation (ABMT) in patients with acute myeloid leukemia (AML) in untreated first relapse (REL1) or in second complete remission (REM2). PATIENTS AND METHODS Forty-seven patients with AML in REL1 (n = 21) or in REM2 (n = 26) were treated with busulfan (BU) and cyclophosphamide (CY) with or without total-body irradiation (TBI) followed by ABMT. All REL1 patients and four REM2 patients had marrow stored during first remission (REM1). Twenty-seven had marrow stored with and 20 without treatment in vitro with 4-hydroperoxycyclophosphamide (4-HC). Eighteen patients received BU and CY and 29 received BU, CY, and TBI. REL1 patients relapsed within a median of 9 months (range, 2 to 26) after marrow harvest and were transplanted a median of 30 days (range, 9 to 87) from detection of relapse. RESULTS With a median follow-up of 2.1 years (range, 0.4 to 5.3), 19 patients survive in remission (10 of 21 in REL1; nine of 26 in REM2). The actuarial probabilities of relapse-free survival at 2 years for patients transplanted in REL1 and REM2 were 45%+/- 22% and 32%+/- 18%, respectively (P =.33). The corresponding probabilities of relapse were 30%+/- 26% and 44%+/- 23%, respectively (P =.45). No conclusions could be drawn about the benefits of adding TBI to BU plus CY. There were no significant differences in neutrophil or platelet recovery or in posttransplant probabilities of relapse and nonrelapse mortality between patients who received marrow treated or not treated with 4-HC. CONCLUSION These results suggest that ABMT may produce long-term leukemia-free survival in approximately one third of patients with AML in REL1 or in REM2. There is no apparent clinical advantage in attempting to obtain second remissions in relapsed patients before ABMT if marrow has been cryopreserved during REM1. Although a strategy of transplantation in REL1 has advantages for the patient, such an approach involves the storage of marrow, which may not be used, and is impractical without the coordinated support of the treating physician, the patient, and the marrow transplant center.
Treatment of refractory acute nonlymphocytic leukemia with a combination of pyrazofurin and cytarabine.
Six patients with refractory acute nonlymphocytic leukemia were treated with pyrazofurin (15 mg/m2) followed by cytarabine (100 mg/m2) every 12 hours for 6--21 days. All patients cleared their peripheral blood of blast cells but no complete remissions were achieved. Excessive toxicity to skin and mucous membranes was observed. In the doses used, this combination is too toxic for further use. Alternate treatment schedules should be explored.
Latest similar papers:
Masayuki Nagasawa, Daisuke Tomizawa, Youichirou Tsuji, Michiko Kajiwara, Tomohiro Morio, Shigeaki Nonoyama, Minoru Asada, Shuki Mizutani
Department of Pediatrics and Developmental Biology, Graduate School, Tokyo Medical and Dental University, 5-45, Yushima 1-chome, Bunkyo-ku, Tokyo 113-8519, Japan. email@example.com
Monosomy 7 syndrome in infant is considered as pre-leukemic condition of poor prognosis. However, it seems controversial recently, because some cases of monosomy 7 syndrome showed spontaneous remission. We report 2-year-old girl with severe pancytopenia, who presented with monosomy 7. Morphologically, there was little dysplasia in the trilineage hematopoiesis. Monosomy 7 clone of CD34 positive cells, bone marrow mononuclear cells (BMMNC), and peripheral nuclear cells was 4.0, 40, and 3.8%, respectively. Immunosuppressive therapy was effective along with the disappearance of monosomy 7 clone. WT1 mRNA expression was not increased in monosomy 7 clone. Pathogenesis of monosomy 7 and its relation to aplastic anemia is discussed.
Hematol J. 2003 ;4 (4):292-4 12872156
Virological, serological and haemopoietic colony studies and its correlation with the outcome of severe aplastic anemia.
Kanjaksha Ghosh, Yegneshwar Iyer, Atanu Basu, Manisha Madkaikar, Farah Jijina, Umapathy Shankarkumar, Dipika Mohanty
Successful cytokine treatment of aplastic anemia following living-related orthotopic liver transplantation for non-A, non-B, non-C hepatitis.
S Sato, S Fuchinoue, M Abe, K Kitajima, T Tojimbara, I Nakajima, T Agishi, H Shiraga, K Ito, K Takasaki, E Hashimoto, N Hayashi, K Tanaka
Department of Surgery III, Tokyo Womens Medical College, Japan.
The assessment of the hematopoietic reservoir after immunosuppressive therapy or bone marrow transplantation in severe aplastic anemia.
M Podesta, G Piaggio, F Frassoni, A Pitto, P Zikos, M Sessarego, M Abate, M Teresa Van Lint, G Berisso, A Bacigalupo
Divisione Ematologia 2, Ospedale S. Martino, Genova, Italy.
We investigated the hematopoietic reservoir in 43 severe aplastic anemia (SAA) patients following immunosuppression (IS)(n = 15) or bone marrow transplantation (BMT)(n = 28), at a median interval of 5 years (range, 2-20) from treatment. All patients had normal blood counts, good marrow cellularity, and normal numbers of colony forming unit-granulocyte macrophages (CFU-GM). Burst forming unit-erythroid (BFU-E) and colony forming unit-granulocyte erythroid megakaryocyte macrophages (CFU-GEMM) numbers were reduced when compared with normal controls. However, the most pronounced defect was observed at the level of long-term culture-initiating cells (LTC-IC), which significantly differed from controls (P <.00001) both for IS and BMT patients. Their number did not improve with time and was not affected by transplant or treatment-related variables. When IS patients were compared with BMT we found comparable numbers of CFU-GEMM (P =.8) and LTC-IC (P =.9), but lower numbers of BFU-E and CFU-GM (P =.05 and P =.004, respectively), suggestive of a persistent suppressive mechanism. These data indicate that LTC-IC numbers are severely reduced in BMT and IS patients, contradicting the common belief that the former are fully reconstituted as compared with the latter. In addition, the number of mature cells and committed progenitors does not seem to reflect the real size of the hematopoietic reservoir and few stem cells may be sufficient to guarantee normal hematopoiesis long term.
Aetiology of severe aplastic anaemia and outcome after allogeneic bone marrow transplantation or immunosuppression therapy. Working Party on Severe Aplastic Anaemia of the European Blood and Marrow Transplantation Group.
Divisione Ematologia 2, Ospedale San Martino, Genova, Italy.
This study is based on 2163 patients with severe aplastic anaemia from the European Bone Marrow Transplantation Working Party Registry on Severe Aplastic Anaemia. It was designed to define patients' characteristics according to different aetiology and to assess the impact of aetiology of the aplasia on treatment outcome. Significant differences were found in age, sex, disease severity and DR typing between idiopathic, post-hepatitis and drug-associated severe aplastic anaemia. These differences did not affect the overall outcome of patients treated with bone marrow transplantation (BMT) or immunosuppression (IS) therapy.
Immunosuppressive therapy in bone marrow aplasia: the stroma functions normally to support hematopoiesis.
University of Cape Town Leukaemia Centre, South Africa.
In aplastic anemia (AA) patients responsive to antilymphocyte globulin (ALG) therapy, abnormalities in both stroma and progenitor cell (PC) pool have been described. The relevance of each pathophysiologic defect was characterized in 16 individuals, and data were compared to results from seven normal volunteers. Bone marrow mononuclear cells were split into two fractions. Stromal layers (SL) were prepared from the first, and a CD34+ enriched population was obtained by immunomagnetic selection from the second. In cross-culture experiments, 1 x 10(4) of the latter from patients or controls were seeded on preformed SL, and adhesive PC were scored for the formation of blast colonies (CFU-Bl) on day 5 of culture. Nonadherent progenitors were recovered and quantitated in a standard clonogenic assay (CFU-GM). There were significantly fewer CD34+ cells in the AA group (median 0.65%, SD 0.39%, vs. 1.62%, SD 1.4%; p = 0.002). No morphological or cytologic differences between normal and aplastic SL were detected. Both equally supported the growth of CFU-Bl from normal progenitors (mean 117, SD 20.4, and 103.1, SD 30.4), while this value was reduced for the aplastic PC (mean 41.06, SD 42.9; p = 0.0002, exact two-tailed test). Similarly, the AA nonadherent PC had a decreased CFU-GM growth (mean 142.6, SD 104.8, vs. mean 361.7; SD 91.3), with a lower total clonogenic output (p = 0.0009). We conclude that aplastic stroma appropriately supports the growth of normal progenitors, whereas the depressed clonogenicity of the corresponsing population derived from AA is unrelated to their attachment to SL but intrinsic to the CD34+ cells, whether adherent or not.
Malignant tumors occurring after treatment of aplastic anemia. European Bone Marrow Transplantation-Severe Aplastic Anaemia Working Party.
G Socié, M Henry-Amar, A Bacigalupo, J Hows, A Tichelli, P Ljungman, S R McCann, N Frickhofen, E Van't Veer-Korthof, E Gluckman
Hôpital Saint Louis, Paris, France.
BACKGROUND AND METHODS. Recent studies have shown that long-term survivors of acquired aplastic anemia may be at high risk for malignant diseases. We assessed the risk of cancer after aplastic anemia was treated with immunosuppression or bone marrow transplantation and sought to identify risk factors according to treatment. The study population consisted of 860 patients treated by immunosuppression and 748 patients who had received bone marrow transplants for the treatment of severe aplastic anemia. The risk of cancer was analyzed overall and according to treatment relative to the risk in the general population. In calculating relative risk, we excluded patients with myelodysplastic syndromes or acute leukemias arising less than 6 months after treatment, and solid cancers arising less than 12 months after treatment, because of a possible association with aplastic anemia itself rather than with the treatment received. RESULTS. Forty-two malignant conditions were reported in the 860 patients who received immunosuppressive therapy: 19 cases of myelodysplastic syndrome, 15 cases of acute leukemia, 1 case of non-Hodgkin's lymphoma, and 7 solid tumors. Nine were reported in the 748 patients who received bone marrow transplants: two cases of acute leukemia and seven solid tumors. After the exclusions listed above, the overall relative risk of cancer was 5.50 (P < 0.001) as compared with that in the general European population; the risk was 5.15 (P < 0.001) after immunosuppressive therapy and 6.67 (P < 0.001) after transplantation. The 10-year cumulative incidence rate of cancer was 18.8 percent after immunosuppressive therapy and 3.1 percent after transplantation. The risk factors for myelodysplastic syndrome or acute leukemia after immunosuppressive therapy included the addition of androgens to the immunosuppressive treatment (relative risk = 0.28), older age (relative risk = 1.03), treatment in 1982 or later, as compared with 1981 or earlier (relative risk = 3.01), splenectomy (relative risk = 3.65), and treatment with multiple courses of immunosuppression (relative risk = 2.26). Risk factors for solid tumors after bone marrow transplantation were age (relative risk = 1.11 per year) and the use of radiation as a conditioning regimen before transplantation (relative risk = 9.56); such tumors occurred only in male patients. CONCLUSIONS. Survivors of aplastic anemia are at high risk for subsequent malignant conditions. Myelodysplastic syndrome and acute leukemia tend to follow immunosuppressive therapy, whereas the incidence of solid tumors is similar after immunosuppression and after bone marrow transplantation.
Oral administration of chloramphenicol did not significantly alter the early stage of chronic hypoplastic marrow failure induced by busulfan. Chloramphenicol reduction products were most suppressive to antigen-reactive cells detected by the Mishell-Dutton plaque-forming cell assay. This reduction in immunocompetence may be an important parameter in the initial phases of the etiology of chloramphenicol-induced aplastic anemia.