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Blood Nov 1989Autologous bone marrow transplantation (ABMT) makes it possible to escalate the dose of cytotoxic treatment to a lethal range. Disease-free survival (DFS) following...
Autologous bone marrow transplantation (ABMT) makes it possible to escalate the dose of cytotoxic treatment to a lethal range. Disease-free survival (DFS) following myeloablative therapy and ABMT has been shown to be superior to conventional treatment in high risk patients with acute myelogenous leukemia (AML). It was the purpose of the present study to compare hematopoietic reconstitution, actuarial DFS, and relapse rate of patients transplanted in first complete remission (CR) of AML with those in second or subsequent CR, and to evaluate transplant related mortality. Fifty-two patients with AML, 22 in first CR (low risk) and 30 in second or subsequent CR (high risk), underwent total body irradiation (12.1 to 16.7 Gy) and cyclophosphamide (CY) treatment (200 mg/kg) followed by ABMT. The autograft was incubated with the active CY derivative Mafosfamide (ASTA Werke, Bielefeld, Federal Republic of Germany) to reduce the number of possibly contaminating clonogenic tumor cells. All patients showed three lineage engraftments with platelet recovery observed as being the slowest. The transplant related death rate was low at 5.8%. There was no significant difference in the kinetics of polymorphonuclear (PMN) cell or platelet reconstitution between the low and high risk patient subgroups. The estimated probability of DFS (relapse) after ABMT in first CR was 61% (36%) compared with 34% (65%) in second or subsequent CR, the longest follow-up being 55 months and 57 months, respectively (median follow-up 31 months and 19 months, respectively). ABMT offers a stable long-term DFS when performed in first CR with no relapses occurring in over a year after transplantation. Six later relapses, however, were seen after ABMT in second or subsequent CR, although DFS was not statistically different from that of first remission patients (P = .72).
Topics: Adult; Bone Marrow; Bone Marrow Transplantation; Cyclophosphamide; Hematopoiesis; Humans; Leukemia, Myeloid, Acute; Middle Aged; Survival Rate; Transplantation, Autologous
PubMed: 2804340
DOI: No ID Found -
International Journal of Cell Cloning Jul 1989Human acute myelogenous or lymphoblastic leukemia cells of the K-562 and CCRF-SB lines were mixed with an excess of normal human bone marrow cells to simulate a leukemia...
Limiting dilution analysis for detection of residual leukemic cells after bone marrow combined decontamination with mafosfamide followed by merocyanine-540-mediated photosensitization.
Human acute myelogenous or lymphoblastic leukemia cells of the K-562 and CCRF-SB lines were mixed with an excess of normal human bone marrow cells to simulate a leukemia remission marrow. The cell mixtures were then incubated in vitro with mafosfamide (AZ) followed by the photoreactive dye merocyanine-540 (MC). Treated cells (1 x 10(4] were seeded in microwell plates, and increasing numbers of the line used to contaminate the normal marrow were added. Treatment with AZ alone produced total elimination (i.e., 6 logs) of CCRF-SB cells, while addition of merocyanine-540 increased the cloning efficiency from 22% to 24.4%. After treatment of the K-562-contaminated cell mixtures with AZ, nearly 1.6 logs of K-562 acute myelogenous blasts were still present, whereas AZ purging followed by MC-mediated photosensitization resulted in 100% elimination of clonogenic cells. Moreover, the combined treatment caused an increase of the cloning efficiency from 37.3% to 62%, clearly indicating that cleansing by the two agents combined was more effective than treatment with one agent alone.
Topics: Antineoplastic Agents; Bone Marrow; Cell Line; Colony-Forming Units Assay; Cyclophosphamide; Dye Dilution Technique; Humans; Leukemia; Light; Pyrimidinones; Radiation-Sensitizing Agents; Tumor Stem Cell Assay
PubMed: 2671163
DOI: 10.1002/stem.5530070404 -
Cancer Immunology, Immunotherapy : CII 1989By cloning in vitro we have obtained two sublines of the L5222 rat leukemia, one with high (L5222-S) and the other with low (L5222-R) in vivo sensitivities to non-toxic...
By cloning in vitro we have obtained two sublines of the L5222 rat leukemia, one with high (L5222-S) and the other with low (L5222-R) in vivo sensitivities to non-toxic doses of mafosfamide, a stabilized derivative of 4-hydroxy-cyclophosphamide. This sensitivity in vivo was not related to the cytotoxic activity of the drug in vitro. Treatment of rats bearing the L5222-S and of mice transplanted with the MOPC-315 plasmocytoma with low doses of mafosfamide or cyclophosphamide resulted in a high percentage of surviving animals, which were resistant to a subsequent tumor challenge. Viable leukemic cells were needed to establish antitumor immunity, since it was not possible to induce resistance by injection of mitomycin-C-treated, non-viable L5222 cells. The adoptive transfer of spleen cells from animals immune against the L5222-S and the MOPC-315 resulted in resistance of the syngeneic recipients against a rechallenge with tumor cells, provided that the animals were treated with an immunosuppressive dose (100 mg/kg) of cyclophosphamide prior to the spleen cell implantation. In nude mice treatment of the L5222 with low doses of mafosfamide also resulted in surviving animals, however resistance to a second tumor challenge occurred only sporadically. The data presented confirm that therapy with cyclophosphamide or mafosfamide enhances host antitumor immunity but, contrary to previous reports, it could be demonstrated that successful tumor rejection was independent of T cells.
Topics: Adjuvants, Immunologic; Animals; Antigens, Neoplasm; Clone Cells; Cyclophosphamide; Drug Administration Schedule; Female; Immunity, Innate; Immunization, Passive; Leukemia, Experimental; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Rats; Rats, Inbred Strains
PubMed: 2924329
DOI: 10.1007/BF00204986 -
Cancer Immunology, Immunotherapy : CII 1989The influence of cis-4-sulfoethylthio-cyclophosphamide (mafosfamide) on natural killer cell activity was examined in vitro in order further to elucidate the possible...
The influence of cis-4-sulfoethylthio-cyclophosphamide (mafosfamide) on natural killer cell activity was examined in vitro in order further to elucidate the possible immunological mechanisms of tumor regressions following low-dose oxazaphosphorine therapy. It was observed that cytotoxicity of human blood lymphoid cells was unchanged or reduced when the lymphocytes were pretreated for 24 h with mafosfamide or when the drug was present during incubation with K562 cells. However, when lymphoid cells were preincubated with human leukocyte interferon plus mafosfamide, natural killer activity was enhanced above the level caused by interferon alone. This enhancement was noted at mafosfamide concentrations of 1 nM-1 microM and was only present when the lymphocyte preparation was contaminated with monocytes.
Topics: Adjuvants, Immunologic; Cyclophosphamide; Cytotoxicity, Immunologic; Dose-Response Relationship, Immunologic; Humans; Interferon Type I; Killer Cells, Natural; Monocytes
PubMed: 2720708
DOI: 10.1007/BF00199291 -
Cancer Immunology, Immunotherapy : CII 1989Some conventional and experimental anti-cancer drugs were tested for their effect on concanavalin-A-induced interferon gamma release from rat splenocytes in vitro. When...
Some conventional and experimental anti-cancer drugs were tested for their effect on concanavalin-A-induced interferon gamma release from rat splenocytes in vitro. When 2.5 x 10(6) rat splenocytes/ml, stimulated with 1 microgram/ml concanavalin A, were incubated with various non-cytotoxic doses of the vinca alkaloid vincristine, there was an inhibition of the release of interferon gamma in culture supernatants. The antitumour antibiotics bleomycin and Adriamycin, alkylating agents 4-hydroperoxycyclophosphamide and mafosfamide, and the immunoactive peptides FK156 and FK565 did not affect the release of interferon gamma under similar conditions. However, cyclosporin A, in similar experiments, markedly inhibited the release of interferon gamma.
Topics: Animals; Antineoplastic Agents; Bleomycin; Concanavalin A; Cyclosporins; Diaminopimelic Acid; Doxorubicin; In Vitro Techniques; Interferon-gamma; Male; Oligopeptides; Rats; Rats, Inbred Lew; Vincristine
PubMed: 2480848
DOI: 10.1007/BF01665007 -
Blood Nov 1987To stimulate a leukemia remission marrow, cell suspensions of normal human bone marrow were mixed with human acute lymphoblastic or myelogenous leukemic cells of the...
To stimulate a leukemia remission marrow, cell suspensions of normal human bone marrow were mixed with human acute lymphoblastic or myelogenous leukemic cells of the CCRF-SF, Nalm-6, and K-562 lines. The cell mixtures were incubated in vitro with mafosfamide (AZ) or with the photoreactive dye merocyanine 540 (MC-540). A quantity of 10(4) cells of the treated suspensions was dispensed into microculture plates, and graded cell numbers of the line used to contaminate the normal marrow were added. Limiting-dilution analysis was used to estimate the frequency of leukemia cells persisting after treatment with the decontaminating agents. Treatment with AZ or MC-540 produced a total elimination (ie, 6 logs or 5.3 logs respectively) of B cell acute leukemia cells (CCRF-SB), whereas nearly 1.7 logs and 2 logs of K-562 acute myelogenous blasts were still present in the cell mixtures after treatment with MC-540 and AZ, respectively. Treatment of the Nalm-6-contaminated cell mixtures with AZ resulted in 100% elimination of clonogenic cells, whereas nearly 80% decontamination was obtained with MC-540. Our results suggest that treatment with AZ could be an effective method of eliminating clonogenic tumor cells from human bone marrow. MC-540, shown by previous studies to spare sufficient pluripotential stem cells to ensure hemopoietic reconstitution in the murine model and in clinical application, has comparable effects and merits trials for possible clinical use in autologous bone marrow transplantation.
Topics: Bone Marrow; Bone Marrow Cells; Cell Line; Cyclophosphamide; Humans; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Pyrimidinones; Radiation-Sensitizing Agents
PubMed: 3478102
DOI: No ID Found -
British Journal of Cancer Jul 1986The potential use of human head and neck (H & N) tumours, growing in athymic nude mice, for preclinical assessment of cytostatic drug sensitivity in a soft agar cloning...
The potential use of human head and neck (H & N) tumours, growing in athymic nude mice, for preclinical assessment of cytostatic drug sensitivity in a soft agar cloning system was examined. Of 20 H & N tumour xenografts, obtained from 6 different xenograft lines, 17 demonstrated sufficient colony growth to evaluate in vitro drug sensitivity. Moreover, all xenografts provided enough cells to test 8 cytostatic drugs at 3 concentrations each. A dose-dependent inhibition of colony growth was obtained with all drugs tested, except methotrexate. Tumours were considered sensitive when the drug concentration required to inhibit colony formation by 50%, was less than 1/10 of the peak plasma concentration in patients. All H & N tumour lines were resistant to cisplatin, doxorubicin, hydroxyurea, mafosfamide (an in vitro active analogue of cyclophosphamide) and methotrexate. Bleomycin was active in 1/6 and 5-fluorouracil in 6/6 of the H & N tumour lines tested. In 32 cases the in vitro data of the H & N tumour lines and a chemosensitive rat rhabdomyosarcoma were compared directly with in vivo results obtained in nude mice. The clonogenic assay correctly predicted sensitivity in 4/6 (66.7%) and resistance in 21/26 (80.8%) of the cases. A lack of correlation was noted for methotrexate, 5-fluorouracil and cyclophosphamide. In vitro culture of human H & N xenografts may provide a means for a rapid and large scale screening to identify new drugs active against H & N malignancies. In addition the clonogenic assay may help to select drugs for subsequent testing in the nude mouse xenograft model. The lack of correlation for some drugs in the present study indicates that there are some limitations in the use of xenograft tumour material for in vitro testing of new drugs.
Topics: Animals; Cisplatin; Cyclophosphamide; Dose-Response Relationship, Drug; Doxorubicin; Drug Resistance; Female; Fluorouracil; Head and Neck Neoplasms; Humans; Hydroxyurea; In Vitro Techniques; Methotrexate; Mice; Mice, Nude; Neoplasm Transplantation; Rats; Tumor Stem Cell Assay
PubMed: 3730256
DOI: 10.1038/bjc.1986.151 -
Blood May 1986The sensitivity of human myeloblastic leukemic (CFU-L) and normal hemopoietic stem cells (CFU-GM and BFU-e) to Asta Z 7557 (INN Mafosfamide) was studied with regard to... (Clinical Trial)
Clinical Trial
The sensitivity of human myeloblastic leukemic (CFU-L) and normal hemopoietic stem cells (CFU-GM and BFU-e) to Asta Z 7557 (INN Mafosfamide) was studied with regard to autologous bone marrow transplantation (ABMT) with cleansed marrow for consolidation therapy in adult patients with acute leukemia (AL) in remission. Establishment of the dose-response curves for CFU-GM (n = 37), BFUe (n = 11), and myeloblastic CFU-L (n = 9) demonstrated a wide range of sensitivity from patient to patient for all three progenitors. Whereas CFU-L, CFU-GM, and BFU-e grown in semisolid cultures disclosed similar sensitivities to Asta Z 7557, long-term culture (LTC) studies (n = 41) indicated a higher resistance of early progenitors. In an effort to achieve a maximum tumor cell kill and yet spare a sufficient amount of normal stem cells to ensure consistent engraftment, we defined the optimal dose for marrow cleansing as the dose sparing 5% CFU-GM (LD95). This dose was established from a preincubation test (PIT) realized on a 10-mL marrow aspirate taken 15 days before marrow collection in each individual patient. Twenty-four adult patients while in remission of AL (20 in complete remission, four in partial remission) were consolidated by cyclophosphamide 60 mg/kg X 2 and total body irradiation at 10 Gy followed by ABMT with marrow cleansed by Asta Z 7557 according to the specification described above. Patients were divided in two groups: group 1, unfavorable prognosis (11 patients); group 2, standard prognosis [13 patients in first complete remission (CR)]. All patients engrafted on leukocytes (median day for recovery to 10(9)/L: day 30), patients with ALL recovered faster than patients with ANL (median day 19 v 34). Similarly, recovery of platelets to 50.10(9)/L occurred sooner in patients with ALL (median day 67, range day 23 through 90) whereas three patients with acute nonlymphoblastic leukemia (ANLL) in group 2 had to be supported with platelet transfusions for more than one year. In group 1, six patients had recurrent tumor within six months; three patients died from toxicity with no evidence of tumor. Two patients are still disease-free with a short follow-up (nine and ten months). In group 2, two patients died from toxicity with no evidence of leukemia three and 16 months post-ABMT. One patient with a M5 ANLL and one patient with ALL relapsed at six and 15 months, respectively. Nine patients have remained in CR or are disease-free with a median follow-up of 22 months.(ABSTRACT TRUNCATED AT 400 WORDS)
Topics: Adolescent; Adult; Bone Marrow Transplantation; Cell Separation; Clinical Trials as Topic; Colony-Forming Units Assay; Cyclophosphamide; Erythroblasts; Female; Granulocytes; Humans; Leukemia; Liver; Male; Middle Aged; Stem Cells
PubMed: 3516254
DOI: No ID Found