Adolescent ; Adult ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Bone Marrow Purging ; Female ; Humans ; Lymphoma, B-Cell ; therapy ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Retrospective Studies ; Rituximab ; Transplantation, Autologous ; Treatment Outcome ; Young Adult

The objective of this study was to investigate the effect of a novel Zinc phthalocyanine (ZnPcH(1)) based photodynamic therapy (PDT) on acute monocytic leukemia cell lines SHI-1 and its mechanism, so as to provide theory basis for bone marrow purging in vitro for patients with leukemia. The killing effect of ZnPcH(1)-PDT on SHI-1 cells were assessed by MTT method; the SHI-1 cell death patterns were analyzed by AO/EB fluorescence staining, TdT-mediated dUTP nick end labeling (TUNEL), DNA ploidy analysis, and Annexin V-FITC/PI double staining.Cell mixture was established by integrating SHI-1 cells with normal bone marrow MNC (by 1:100-1:10 000). Purging effect of ZnPcH(1)-PDT against SHI-1 mixed into normal MNC was assessed by analyzing the expression of fusion gene MLL/AF6 mRNA using nested RT-PCR. The results showed that ZnPcH(1)-PDT could effectively inhibit SHI-1 cell proliferation in dose-dependent manner, and ZnPcH(1)-PDT could induce cell apoptosis in time-dependent manner. 0.5 µmol/L ZnPcH(1)-PDT could completely photoinactivated kill SHI-1 cells in the simulated remission bone marrow. It concluded that ZnPcH(1)-PDT may be a effective and convenient promising purging technique for leukemia.
Apoptosis ; drug effects ; Bone Marrow Purging ; methods ; Cell Death ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Indoles ; pharmacology ; therapeutic use ; Leukemia, Monocytic, Acute ; drug therapy ; pathology ; Organometallic Compounds ; pharmacology ; therapeutic use ; Photochemotherapy ; Photosensitizing Agents ; pharmacology ; therapeutic use

The objective of this study was to explore the occurrence and clinical features of acute graft versus host disease (aGVHD) in non-myeloablative stem cell transplantation (NAST). 19 cases developed aGVHD out of 71 cases with NAST in recent years were analyzed retrospectively. Out of 19 cases, 9 males and 10 females at the median age of 38 (18-59), 16 cases with grade I-II aGVHD, 3 cases with grade III-IV aGVHD. The results indicated that the incidence of aGVHD in NAST was 26.7% (19/71), and severe aGVHD was 4.2%, the median onset time was 58 days (17-240 days) after transplantation. Skin and especially the intestine were the main target organs of aGVHD, while diarrhea occurred as the first symptom in 7 cases, 3 cases showed mixed acute and chronic GVHD involving more locations at the same time. aGVHD occurrence was 38.2% in those patients with full donor chimerism (FDC) and 16% in patients with the mixed chimerism (MC). It is concluded that aGVHD in NAST is less in occurrence, lighter in severity and later in time, but higher occurrence in those with early FDC, which intestine and skin are the main target organs. The clinical course is prolonged and easily complicated with severe infection in the later phase. Early combined therapy with powerful supportive treatment is necessary.
Adolescent ; Adult ; Bone Marrow Purging ; China ; epidemiology ; Female ; Graft vs Host Disease ; epidemiology ; etiology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Incidence ; Leukemia ; therapy ; Male ; Middle Aged ; Young Adult

OBJECTIVETo establish a murine transplant model for bone marrow purging of metastatic breast cancer and to explore the efficiency of Econazole (Ec) as a purging agent.

METHODSMixtures of TSA /Neo breast cancer cells and murine bone marrow cells were transplanted into lethally irradiated mice following purging with Econazole or saline in vitro. The recipient mice were monitored for hematopoietic engraftment, appearance of metastatic nodules in lungs and the overall survival.

RESULTSAll the mice receiving i.v. injection of TSA cells developed metastatic lung nodules. The hematological recovery was not delayed in mice transplanted with Ec purged bone marrow. More importantly, metastatic lung nodules were not seen in Ec treated group and the overall survival was improved.

CONCLUSIONThe purged metastatic breast cancer cell bone marrow transplant model was easily established and reproducible. Ec could be used to purge the bone marrow grafts contaminated with breast cancer cells.

Animals ; Antineoplastic Agents ; pharmacology ; Bone Marrow Purging ; Bone Marrow Transplantation ; Cell Line ; Econazole ; pharmacology ; Female ; Mammary Neoplasms, Experimental ; pathology ; Mice ; Mice, Inbred BALB C

Adenoviridae ; genetics ; Animals ; Bone Marrow Purging ; methods ; Genes, p53 ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; virology ; Humans ; Receptors, Virus ; genetics ; Transplantation, Autologous ; methods

BACKGROUNDAn effective purging technique plays an important role in autologous hematopoietic stem cells transplantation. Photodynamic therapy (PDT) provides a novel approach for this purpose. This study dealt with the purging effects of di-sulfo-di-phthalimidomethyl phthalolcyanine zinc (ZnPcS2P2)-based photodynamic therapy (ZnPc-PDT).

METHODSFluorescence intensity of cell extracts was measured using a fluorescence spectrophotometry. The proliferative potency of K562 cells and HL60 cells was detected using MTT colorimetric assay, Typan blue dye exclusion method, colony formation test. The proliferative potency of normal hematopoietic cells was evaluated using mixture colony-forming unit (CFU-Mix), granulocyte-macrophage colony-forming unit (CFU-GM), and erythrocyte colony-forming unit (CFU-E) assays. K562 cells were mixed with normal mononuclear cells (MNCs) at ratios of 1:100 and 1:1000 for creating the model of simulated remission bone marrow. Colony formation test and nested-RT-PCR were carried out to detect the residual K562 cells in cell mixture.

RESULTSAfter a 5-hour incubation with ZnPcS2P2, the content of ZnPcS2P2 in normal MNCs was the lowest value. At the same time, the content in K562 cells and HL60 cells was very high. Therefore, the time point was selected as the optimal one for irradiating the cell suspensions. ZnPc-PDT could significantly kill proliferative K562 cells and HL60 cells in a dose-dependent manner. At the concentration of 1.0 microg/ml, the inhibitory rate of ZnPc-PDT on the colony formation was 100% for K562 cells, 89.7% for HL60 cells. 0.25 microg/ml ZnPc-PDT could completely photoinactivate residual K562 cells in the simulated remission bone marrow. Under an identical condition, the inhibitory rates of CFU-Mix, CFU-GM, CFU-E were 18.0%, 18.6%, and 17.8% respectively.

CONCLUSIONZnPc-PDT appears to be a promising approach for bone marrow purging.

Bone Marrow Purging ; Bone Marrow Transplantation ; Cell Division ; drug effects ; Colorimetry ; HL-60 Cells ; Hematopoietic Stem Cells ; drug effects ; Humans ; Indoles ; pharmacology ; K562 Cells ; Photochemotherapy ; Photosensitizing Agents ; pharmacology ; Phthalimides ; pharmacology

OBJECTIVETo evaluate the outcome of patients with de novo acute leukemia (AL, no AML-M(3)) in CR(1) undergone autologous hematopoietic stem cell transplantation (auto-HSCT) or HLA-identical sibling allogeneic HSCT (allo-HSCT).

METHODSForty-six AL patients received allo-HSCT and 94 received auto-HSCT in CR(1). The conditioning regimens mainly consisted of TBICy, BuCy and MAC. Cyclosporine plus methotrexate, or cyclosporine alone, or FK506 alone was used for graft-versus-host disease (GVHD) prophylaxis. Among auto-HSCT group, 39 patients received purged autologous bone marrow and 38 received immunotherapy and/or maintenance chemotherapy after transplant.

RESULTSMyeloid reconstitution was achieved in all patients. After a median of 700 (range, 18 approximately 5563) days follow-up, the probabilities of leukemia-free survival (LFS) at 5 year were not significantly different in these two groups: (51.5 +/- 5.4)% for auto-HSCT group and (52.8 +/- 7.6)% for allo-HSCT group (P > 0.05). There was a lower cumulative relapse incidence (RI) [(26.3 +/- 6.9)% vs. (52.0 +/- 5.5)%, P > 0.05] but a significantly higher cumulative transplant-related mortality (TRM) [(37.6 +/- 7.8% vs. (14.4 +/- 4.1)%, P < 0.05] in the allo-HSCT group than in auto-HSCT group. Among auto-HSCT group, the patients received purged autografts and/or post-transplant therapy had significantly better LFS and lower RI (P < 0.05) than those received unpurged autografts or no post-transplant treatments [5-y LFS: (62.8 +/- 6.8)% and (38.4 +/- 8.4)%; RI: (37.7 +/- 6.8)% and (74.2 +/- 8.7)%, respectively].

CONCLUSIONThe long-term LFS of auto-HSCT was comparable to that of allo-HSCT in the management of patients with AL in CR(1), because autograft purging and post-transplant treatment can significantly decrease relapse of auto-HSCT patients and auto-HSCT has lower therapy-related toxicities.

Acute Disease ; Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Marrow Purging ; Child ; Combined Modality Therapy ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Immunotherapy ; methods ; Kaplan-Meier Estimate ; Leukemia ; therapy ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Remission Induction ; Retrospective Studies ; Transplantation Conditioning ; methods ; Transplantation, Autologous ; Transplantation, Homologous ; Treatment Outcome

To explore the new approach to prevent graft versus host disease (GVHD) by purging ex vivo T lymphocytes of bone marrow graft through Fas-FasL way, FasL-cDNA was transfected into BALB/c mouse bon e marrow cells by liposome ex vivo. The transfected cells were cultured together with BAC (BALB/c x C57BL/6) mouse bone marrow graft. The mixing bone marrow graft was infused into BALB/c mouse recipients after 60Co-gamma irradiation. The mortality, manifestation and pathologic change of GVHD in recipient mice were observed. The CFU-S and Y chromosome from donor mice were detected. The results showed that compared with control group, the mortality in 60 days of the recipients in the experimental group decreased (20% vs 70%, P < 0.01) and the morbidity of GVHD lowered (40% vs 100%, P < 0.01). The CFU-S counts for all groups were at normal level on 20 days after transplantation. The Y chromosome from donor mice was discovered in 70% bone marrow nucleated cells of recipient mice survived over 2 months in the experimental group. It is concluded that mFasL-cDNA transfected mouse bone marrow cells prevent GVHD after culturing together with bone marrow graft, and accelerate hematopoietic reconstitution in recipient mice.
Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Purging ; Bone Marrow Transplantation ; Fas Ligand Protein ; Female ; Genetic Therapy ; Graft vs Host Disease ; Male ; Membrane Glycoproteins ; genetics ; Mice ; Mice, Inbred BALB C ; Transfection

To compare the clinical outcome of autologous peripheral blood stem cell transplantation (APBSCT) and autologous bone marrow transplantation (ABMT) in treatment of patients with acute leukemia in first remission, 41 patients received APBSCT, 17 patients received unpurged ABMT and 30 patients received purged ABMT. The results showed that hematopoietic recovery was significantly earlier after APBSCT than that after purged or unpurged ABMT. The 3-year disease-free survival (DFS), relapse rate (RR) and transplant-related mortality (TRM) for all patients of 3 groups were 51.7%, 41.7% and 6.8%, respectively. DFS and RR were significantly influenced by disease types (ALL or AML) and intervals between diagnosis and CR(1) or CR(1) and transplant. The main causes of transplant-related death were infection and hemorrhage. After APBSCT, DFS, RR and TRM were 48.4%, 43.9% and 4.9%, respectively, and did not differ significantly from those found in unpurged ABMT (47.1%, 45.6% and 11.8%) or purged ABMT (66.5%, 29.6% and 6.7%). It is concluded that the clinical outcome of APBSCT is similar to unpurged or purged ABMT but APBSCT allows faster recovery of hematopoiesis and needs less transfusion support.
Acute Disease ; Adolescent ; Adult ; Bacterial Infections ; etiology ; mortality ; Bone Marrow Purging ; Bone Marrow Transplantation ; adverse effects ; Child ; Disease-Free Survival ; Female ; Follow-Up Studies ; Hemorrhage ; etiology ; mortality ; Humans ; Leukemia ; pathology ; therapy ; Leukemia, Erythroblastic, Acute ; pathology ; therapy ; Leukemia, Monocytic, Acute ; pathology ; therapy ; Leukemia, Myeloid, Acute ; pathology ; therapy ; Leukemia, Myelomonocytic, Acute ; pathology ; therapy ; Leukemia, Promyelocytic, Acute ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Peripheral Blood Stem Cell Transplantation ; adverse effects ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; pathology ; therapy ; Remission Induction ; Survival Rate ; Transplantation, Autologous

Bcr-abl antisense oligodeoxynucleotides (AS-ODNs) have provided evidence of an antileukemia effect when tested in vitro against Philadelphia-positive cells. In order to investigate the efficacy of AS-ODNs as purging agents in chronic myeloid leukemia (CML) patients, K562 cells, a human CML cell line, were treated in vitro with various types of AS-ODNs and interferon-alpha. Cells were treated in vitro for 0 and 36 hr with 40 microgram/mL of AS-ODNs, respectively, and incubated at 37 degrees C for 36 hr. Cytotoxic effects were measured by counting the number of viable cells as well as by MTT test. Clonogenic activities were evaluated by methylcellulose culture for 2 weeks. The effects of purging agents on the rearrangement of bcrabl gene were evaluated by RT-PCR. AS-ODNs inhibited the proliferation of K562 cells with time in cell count assay and MTT test. AS-ODNs were superior to INF-alpha in inhibiting clonogenic activity (recovery rate; 26.3% vs 64.0%). After incubation with bcr-abl AS-ODNs primers and mRNA isolated from K562 cells, positive bands were abolished, especially of b3a2 type and phosphorothioate type. Our results suggest that AS-ODNs mediated purging may be one of the efficient methods and that autograft may be an alternative treatment for allograft in high-risk group patients of CML if they do not have a stem cell donor.
Bone Marrow Purging* ; Colony-Forming Units Assay ; Fusion Proteins, bcr-abl/genetics ; Fusion Proteins, bcr-abl/metabolism ; Hematopoietic Stem Cells/physiology* ; Human ; Leukemia, Myeloid, Chronic/therapy ; Neuroblastoma/therapy* ; Oligonucleotides, Antisense/metabolism* ; Oligonucleotides, Antisense/therapeutic use ; Transplantation, Autologous* ; Tumor Cells, Cultured