To investigate the antitumor activities of the immunoconjugates composed of anti-type IV collagenase monoclonal antibody Fab' fragment and lidamycin (LDM) prepared with different linkers. The immunoconjugates were prepared by linking Fab' to lysine-69 of LDM apoprotein by SPDP, LCSPDP, SMBS or SSMPB as the intermediate drug linkers. Immunoreactivities of the conjugates were determined by ELISA. The cytotoxicities of the conjugates were examined by clonogenic assay. In vivo antitumor effects of the conjugates were evaluated in nude mice bearing subcutaneously implanted HT-1080 tumor. ELISA assay showed that the conjugates retained part of the immunoreactivity of 3G11 against the antigen. The cytotoxicities of the Fab'-SMBS-LDM and Fab'-SSMPB-LDM to HT-1080 cells were significantly potent, compared with Fab'-SPDP-LDM, Fab'-LCSPDP-LDM and free LDM. In animal models at the same condition, free LDM, Fab'-SPDP-LDM and Fab'-LCSPDP-LDM inhibited the growth of HT-1080 tumor by 70.9%, 74.8% and 72.3%, while Fab'-SMBS-LDM and Fab'-SSMPB-LDM reached 78.0% and 87.7%, respectively. The median survival time of the mice treated with free LDM, Fab'-SPDP-LDM and Fab'-LCSPDP-LDM were prolonged by 71.9%, 82.2% and 107.5%, respectively, compared with that of untreated group. Whereas, the median survival time of Fab'-SMBS-LDM and Fab'-SSMPB-LDM were prolonged by 145.2% and 165.8%, respectively, indicating that Fab'-SSMPB-LDM was more effective than Fab'-SMBS-LDM in tumor suppression and life span prolongation. Fab'-SSMPB-LDM has more marked selective antitumor efficacy and lower toxicity, and might be a novel candidate for cancer therapy.
Aminoglycosides ; pharmacology ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antibodies, Monoclonal ; immunology ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Collagenases ; immunology ; Enediynes ; pharmacology ; Fibrosarcoma ; pathology ; Humans ; Immunoconjugates ; pharmacology ; Immunoglobulin Fab Fragments ; immunology ; Matrix Metalloproteinase Inhibitors ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Tumor Burden ; drug effects

OBJECTIVETo evaluate the use of a new reduced intensity of BuCy conditioning regimen for the treatment of malignant hematologic diseases in aged or intolerable patients receiving allogeneic hematopoietic stem cell transplantation (allo-HSCT) from the siblings.

METHODSTwelve patients with acute lymphoblastic leukemia (ALL, n = 4), acute myelogenous leukemia (AML-M(2), n = 2), chronic myelogenous leukemia (CML, n = 4), and myelodysplastic syndromes-refractory anemia with excess blasts (MDS-RAEB, n = 2) were intolerant of conventional myeloablative therapy because of age (older than 50 years) or having severe concurrent diseases. The median age was 49 years (range 42-64 years). Seven were males and five females. Two of the 12 patients were HLA one antigen-mismatched and the rest HLA identical with their donors. The low dosage conditioning regimen consisted of busulfan (2 mg.kg(-1).d(-1) for 3 days), Ara-C (2 g.m(-2).d(-1) for 1 or 2 times), cyclophosphamide (1.0 g.m(-2).d(-1) for 2 days) and anti-T-lymphocyte globulin (ATG 2.5 mg.kg(-1).d(-1) for 4 days, -5 - -2 day). Granulocyte colony-stimulating factor mobilized bone marrow and peripheral blood stem cells (PBSC) were harvested (1 patient using PBSC alone). All patients received cyclosporin A, short-term MTX and mycophenolate mofetil (MMF) for prophylaxis of acute graft-versus-host disease (aGVHD). DNA short tandem repeat (STR) sequence analysis, cytogenetics and molecular-biologic technique were used to analyze chimerism.

RESULTSAll the patients were well tolerated the regimen, with no severe regimen related toxicity. In all the 12 patients, absolute neutrophil count > or = 0.5 x10(9)/L was achieved in 11 to 17 (median 15) days and platelet count > 20 x 10(9)/L in 10 to 23 (median 15) days after transplantation. Complete chimerism was achieved in 11 patients and 1 patient was in mixed chimerism at one month after HSCT. With a median follow-up of 14.5 (4.0-24.0) months, 7 of the 12 patients (58.0%) were alive and 5 (42.0%) of the 7 were disease-free. The probabilities of OS and DFS at 12 months were 75.0% and 48.1%. Five patients (41.6%) had aGVHD and four had local chronic GVHD with a cumulative probability of chronic GVHD of 41.5%.

CONCLUSIONThis reduced intensity conditioning regimen is well tolerated and safe for HSCT in the older patients or patients with severe concurrent medical conditions and can achieve full chimerism and long-term disease-free survival.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; Female ; Hematologic Neoplasms ; drug therapy ; surgery ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Transplantation Conditioning ; methods

OBJECTIVETo retrospectively analyze the results of a consecutive series of 100 ALL patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT) in our center.

METHODSOf the 100 ALL patients, 69 were male and 31 female, with a median age of 29.5 (4 - 47) years. Sixty-nine cases were in the first complete remission (CR(1)), 13 in more than CR(1) and 18 in relapse before transplant. Allo-HSCT from HLA identical siblings was performed for 86 patients, of whom 64 received bone marrow transplantation (BMT) and 22 peripheral blood stem cell transplantation (PBSCT). HLA matched unrelated BMT was performed for 8 patients, cord-blood transplantation from unrelated donor for 6 patients. Forty-five patients underwent allo-HSCT with conditioning regimen of Cy/TBI, 55 with BUCY. Prophylaxis of graft-versus-host disease (GVHD) included long-term MTX regimen (4 cases) and CsA + MTX regimen (96 cases). The average follow-up was 38.1 months.

RESULTSThe 5-year overall survival (OS) and disease-free survival (DFS) of the 100 cases of ALL was 53.4% and 50.5%. The 5-year OS and DFS were significantly longer for patients in CR(1) than in >CR(1) and relapse patients before allo-HSCT (P < 0.001). The outcome of PBSCT seemed superior to that of BMT, but there was no difference between them. Multivariate analysis showed the most significant factor associated with long post allo-HSCT survival was that the patient underwent transplantation in CR(1). There was no significant difference in 5-year OS, DFS, cumulative incidences of relapse rate and treatment related mortality between the two cohorts prepared with TBI or BUCY.

CONCLUSIONSAllo-HSCT can cure a significant proportion of ALL patients, especially for those in CR(1). There was no significant difference in OS, DFS between the two different conditioning regimens and the different transplant choices.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; mortality ; therapy ; Recurrence ; Transplantation, Homologous

OBJECTIVETo investigate the incidence and risk factors of acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (HSCT).

METHODSThe clinical data of 151 cases of allo-HSCT in 150 patients from Nov 2001 to Jan 2004 was analyzed.

RESULTSaGVHD was developed in 60 cases (40.2%), including 43 cases with grade I - II and 17 with grade III - IV aGVHD, the mean time of aGVHD development was 21 days (range 1 - 85 days) after allo-HSCT, 35 out of 55 cases achieved complete response (CR) (63.6%). The early survival rate for grade I - II aGVHD was more than 90%, while that for grade III - IV aGVHD was 46%. Nineteen factors possibly correlated with the development of aGVHD were analyzed. The univariate analysis showed that recipient age, donor's sex, recipient's sex, sex and ABO blood group disparity between donor and recipient, diagnosis, the status of disease, the stage of disease, stem cell source, conditioning regimen (TBI/without TBI), CD34(+) cell number, CD3(+) cell number, early engraftment and neutropenic infection were not closely associated with the occurrence of aGVHD (P > 0.05). On the Cox regression model, 2 independent factors for grade I - IV aGVHD were identified:HLA mismatch (RR = 1.681, P < 0.05) and positive surface antigen (HBsAg) (RR = 1.907, P < 0.05). In addition, the univariate analysis showed aGVHD was strongly associated with CMV infection (P < 0.01).

CONCLUSIONaGVHD is a common complication after HSCT, HLA mismatch and HBsAg positivity are independent risk factors for aGVHD.

Adolescent ; Adult ; Child ; Female ; Graft vs Host Disease ; etiology ; prevention & control ; HLA Antigens ; immunology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Hepatitis B Surface Antigens ; blood ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Transplantation, Homologous ; Young Adult

OBJECTIVETo explore the clinical application of human leukocyte antigen (HLA) mismatched hematopoietic stem cell transplantation (HSCT) for malignant hematological diseases using a new GIAC protocol.

METHODSOne hundred patients with malignant hematological disease received G-CSF mobilization, intensive immunosuppression, ATG and combination of bone marrow and peripheral blood stem cell transplantation at least 1 locus mis-matched hematopoietic stem cell transplant performed with GIAC protocol. The conditioning regimen was intensified and prolonged with combined use of CsA, MMF and ATG for GVHD prophylaxis.

RESULTSAll patients achieved sustained, full donor-type engraftment. The cumulative incidence of grade II approximately IV aGVHD was 48.39%, and grade III approximately IV aGVHD was 12.90%. Thirty-eight patients had cGVHDs which were of extensive type in 11 patients. Twelve patients relapsed, 11 of them were high-risk patients, and 3 returned to CR after donor lymphocyte infusion. Twenty-two patients died, owing to recurrent diseases in 6 and transplant-related complications in 16 cases. Seventy-two patients were alive and disease free, with 1 year disease-free survival probabilities for standard and high risk patients of (83.52 +/- 7.41)% and (47.63 +/- 8.49)%, respectively.

CONCLUSIONThe GIAC protocol for at least 1 locus mismatched hematopoietic stem cell transplantation is relatively safe and efficient for patients with hematological malignancies.

Adolescent ; Adult ; Antilymphocyte Serum ; administration & dosage ; Bone Marrow Transplantation ; Child ; Child, Preschool ; Cyclosporine ; administration & dosage ; Female ; Graft vs Host Disease ; prevention & control ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; Humans ; Immunosuppressive Agents ; therapeutic use ; Leukemia, Myeloid, Acute ; mortality ; therapy ; Male ; Methotrexate ; administration & dosage ; Middle Aged ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; mortality ; therapy ; Transplantation, Homologous

The relationship between Kruppel-like factor 4 (KLF4) and the Notch pathway was determined to investigate the effect of KLF4 on the activation of hepatic stellate cells and underlying mechanisms. Fifty SPF BALB/c mice were randomly divided into two groups. A liver fibrosis model was established in 25 mice as the experimental group, and the remaining 25 mice served as controls. On the day 0, 7, 14, and 35, liver tissues were removed for immunofluorescent detection. The Notch pathway inhibitor DAPT was added to the primary original hepatic stellate cells, and KLF4 and Notch-associated factor expression was detected by qRT-PCR. Additionally, the hepatic stellate cell line LX-2 was used to establish control and experimental groups, and was cultured in vitro. LX-2 cells in the experimental groups were treated with DAPT and the Notch activator transforming growth factor-beta 1 separately, whereas those in the control group were given isotonic culture medium. After 48 h, KLF4 expression was examined by Western blotting. After transient transfection of LX-2 cells to increase KLF4, the expression of Notch factor was examined. Immunofluorescence analysis showed that, with the aggravation of liver fibrosis, the absorbance (A) values of KLF4 were decreased (day 0: 980.73±153.19; day 7: 1087.99±230.23; day 14: 390.95±93.56; day 35: 245.99±87.34). The expression of Notch pathway- related factors (Notch-1, Notch-2, and Jagged-1) in the hepatic stellate cell membrane was negatively correlated to KLF4 expression. With the increase of KLF4 expression, Notch-2 (0.73±0.13) and Jagged-1 (0.43±0.12) expression decreased, whereas Notch-1 level was not detectable. When the Notch pathway was inhibited, KLF4 levels generally increased (18.12±1.31). Our results indicate that KLF4 expression is negatively correlated to the Notch pathway in hepatic stellate cells, which may provide a reference for the treatment of hepatic fibrosis.
Animals ; Cell Line ; Cells, Cultured ; Hepatic Stellate Cells ; metabolism ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Liver Cirrhosis ; metabolism ; Mice ; Mice, Inbred BALB C ; Receptors, Notch ; metabolism ; Signal Transduction ; Transforming Growth Factor beta1 ; metabolism