No abstract available
Animals ; Cell Line* ; Humans* ; Leukemia* ; Methotrexate* ; Mice ; Mice, SCID*

Animals ; Copper ; toxicity ; Disulfiram ; toxicity ; Humans ; Leukemia, Myeloid, Acute ; Mice ; Mice, Inbred NOD ; Mice, SCID

OBJECTIVE: To investigate the tumorigenicity of several multiple myeloma (MM) cell lines transplanted in mice without γ-ray irradiation and to construct the MM disease model to facilitate in vivo experiments. METHODS: NOD/SCID or NSG mice were subcutaneously or caudally transplanted with MM cell lines (LP-1, OPM2, RPMI 8226 and MOLP8), or cell lines with luciferase (RPMI-Luc-Puro, RPMI-Luc-mCherry and MOLP8-Luc-Puro). Tumor growth was observed by measuring the tumor size with a caliper. CD138 tumor cells in peripheral blood were detected by flow cytometry. The free light chain in mouse serum was detected by immunofixation electrophoresis. Tumor type was identified by immunohistochemistry. RESULTS: Twenty one NOD/SCID mice were subcutaneously transplanted with LP-1 cells or OPM2 cells respectively, and no tumors formed till 7 weeks after transplantation. Fifteen NOD/SCID mice subcutaneously transplanted with RPMI 8226 cells showed tumor formation one week later. As of 7 weeks, the rate of tumorigenesis was 80% (12/15). Serum λ light chain was detected and no CD138 tumor cells were detected in peripheral blood. Two NOD/SCID mice each were subcutaneously transplanted with RPMI-Luc-Puro, RPMI-Luc-mcherry and MOLP8-Luc-Puro cells respectively. No tumor signal was detected through IVIS in RPMI-Luc-mcherry cells-transplanted mice. There was tumor signal at 1 week in RPMI-Luc-Puro and MOLP8-Luc-Puro cells-transplanted mice, the former disappeared at 2 weeks and the latter persisted more than 3 weeks. NSG mice subcutaneously transplanted with both cells persistently displayed the tumor signal. Neither NOD/SCID nor NSG mice transplanted with RPMI 8226, RPMI-Luc-Puro, RPMI-Luc-mcherry or MOLP8-Luc-Puro cells through tail vein developed the tumor signal. Only one NSG mice transplanted with MOLP8-Luc-Puro cells appeared transient tumor signal. CONCLUSION Unirradiated mice transplanted with MM cell lines tended to develop local tumor, and failed to develop disseminated tumor. The tumorigenicity of different cell lines is quite different and the vector transfection can reduce the tumorigenic ability. NSG mice with more severe immunodeficiency are more suitable for tumor growth.
Animals ; Carcinogenesis ; Cell Line ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Multiple Myeloma

PURPOSE: Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. METHODS: ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E.coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24, and 48 hr with 8 mm pinhole collimator. RESULTS: An active scFv lym-1 could be produced in E.coli with soluble from using pET vector system. Immunoreactivity and affinity constant of IgG lym-1 were 54% and 1.83 x 10(9) M(-1), respectively, and those of scFv lym-1 were 53.7% and 1.46 x 10(9) M(-1), respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 IgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 IgG lym-1 antibody. CONCLUSIONS: In vitro properties of scFv lym-1 were similar to those of IgG lym-1. ScFv lym-1 showed faster blood clearance than IgG lym-1. These results suggest that scFv lym-1 antibody can be useful for tumor imaging agent.
Animals ; Gamma Cameras ; Immunoglobulin G ; Kidney ; Liver ; Lymphoma* ; Mice ; Mice, SCID ; Periplasm ; Radionuclide Imaging ; Spleen

OBJECTIVEThis study was to establish a stable, effective and reproducible human acute promyelocytic leukemia model in severe combined immunodeficient (SCID) mice by using NB4 cell line, and to investigate the disease course character and biological behaviors.

METHODSThree-five-week-old SCID beige mice were divided randomly into two groups: experimental and control group. SCID mice of experimental group were transplanted by tail vein (iv) injection of 5×10(6) NB4 cells. The WBC cell count and the positive rate of promyelocytes in peripheral blood were dynamically monitored by using smears. Morphological examination and histopathological assay were employed to confirm NB4 cell infiltration in organs (liver, spleen, lung, kidney and brain). The expression level of PML-RARα fusion protein was detected by Western blot.

RESULTSWithin two weeks there was no significant difference in peripheral blood WBC count between two groups (P > 0.05), meanwhile, NB4 cells were not found. At the day 21 and 28 after inoculation, the peripheral blood white blood cell count of experimental group reached to (4.79 ± 1.13)×10(9)/L and (7.62 ± 2.24)×10(9)/L respectively, which were significantly higher than that in control group (P < 0.05); simultaneously, the positive rates of promyelocytes on smears were (2.14 ± 0.63)% and (6.6 ± 2.76)%, respectively. Morphological observation showed single or multiple tumor lumps at day 21 after inoculation; HE staining of tissue biopsies demonstrated a large number of promyelocyte in the liver, spleen, lung, kidney and brain tissue. Cell immunofluorescence results showed that the CD33 expression of bone marrow cells in mice of experimental group was strongly positive (P < 0.05). Western blot confirmed that the PML-RARα fusion protein was expressed variously in liver, kidney and brain tissue.

CONCLUSIONSThe human acute promyelocytic leukemia SCID mouse model is succesfully established by tail vein injection of NB4 cells. This model can mimic the characters of involved bone marrow and diffuse growth of cells. This model is a useful tool to explore the pathogenic mechanism and experimental treatment of human leukemia.

SCID mouse-human leukemia model is an important and useful tool for study on proliferation, differentiation and modulation of leukemic cells. In this article, the establishment of the model, advances in research and application in studies of pathogenesis, cell biology, clinical diagnosis, therapy and assessment of prognosis of leukemia patients are reviewed. The limitations of the model are also commented.
Animals ; Disease Models, Animal ; Humans ; Leukemia ; therapy ; Mice ; Mice, SCID ; Research Design

OBJECTIVETo constitute a model of B immunoblastic lymphomas in the Hu-PBL-SCID mice.

METHODSThe SCID mice were reconstituted by intraperitoneal injection (i.p.) of 5 x 10(7) human lymphocytes from Epstein-Barr virus (EBV) seronegative individuals. After one week, the SCID mice were inoculated with EBV by i.p. injection, and subjected to the investigation of whether there was any tumor in the abdomen of such SCID mice four weeks later. The characteristics of the found tumor was observed by the methods of Hematoxylin-eosin (HE) stain, immunohistochemical staining and polymerase chain reaction (PCR).

RESULTSCompared with the control groups, all the EBV-infected Hu-PBL-SCID mice had abdominal solid tumors [(32 +/- 12.5) mm3] developed, often located in the liver. HE staining and immunohistochemical staining showed the tumors were human B cell lymphomas. EBV DNA could be detected in the tumors by the PCR.

CONCLUSIONSThe model of B immunoblastic lymphomas in the Hu-PBL-SCID mice is successfully constituted, and may well be useful to the human tumor immunological study.

NOD/SCID (non-obese diabetic/severe combined immunodeficient) mice are immune deficient mice which are made by backcross of severe combined immunodeficient mice with non-obese diabetic mice strains. NOD/SCID mice are both innate immune deficiencies and lack of T and B lymphocytes. Various tumor cells can be implanted in this kind of mice, the rejection and graft-versus-host disease (GVHD) occur fewer. Therefore, NOD/SCID mice gradually become a useful tool for the study on Experimental Hematology. This paper comprehensively reviews the biological characteristics of NOD/SCID mice, the establishment of human leukemia model, stem cell transplantation, drug research, deficiency and improvement of NOD/SCID mice in application for study.
Animals ; Disease Models, Animal ; Hematology ; methods ; Leukemia ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Models, Animal ; Research ; trends

OBJECTIVETo compare the tumor-forming rate between the SCID and NOD/SCID mice to set up acute myeloid leukemia (AML) mouse model.

METHODSThe SCID and NOD/SCID mice were injected with HL-60 cells in to the abdominal cavity in order to contruct the AML mouse model. The gereral status of mice was observed, the positive rate of CD33 in bone marrow cells was detected by flow cytometry, the mouse model was identified by pathologic examination.

RESULTSThe tumor-forming rate in constructed model using SCID mouse was 30%, while the tumor-forming rate in constructed model using NOD/SCID mouse was 100%.

CONCLUSIONCompared with SCID mice, the tumor-forming rate in NOD/SCID mice injected with HL-60 cells is high, the incidence of AML is stable, suggesting that the NOD/SCID mouse model is more suitable to explore the pathogenesis of leukemia.

T cell acute lymphoblastic leukemia (T-ALL) is an aggressive leukemia. However the poor prognosis and low morbidity restrict further analysis of the disease. Therefore there is an increasing demand to develop animal models for identifying novel therapeutic approaches. In this study, we inoculated the anti-mouse CD122 monoclonal antibody conditioned NOD/SCID mice with the leukemia cells from 9 T-ALL patients and 1 cell line via the tail vein. Four of the 9 patients and the cell line were successfully engrafted. Flow cytometry detected high percentage of human CD45(+) cells in recipient mice. Immunohistochemistry showed infiltration of human CD45(+) cells in different organs. Serial transplantation was also achieved. In vivo drug treatment showed that dexamethasone could extend survival, which was consistent with clinical observation. These results demonstrated that we successfully established 5 xenotransplantation models of T-ALL in anti-mCD122 mAb conditioned NOD/SCID mice, which recapitulated the characteristics of original disease.
Animals ; Disease Models, Animal ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; Transplantation, Heterologous ; methods