Anti-tumorimmune effects of WT1 polypeptide vaccine on SCID mice with xenografted human monocytic leukemia / 西安交通大学学报(医学版)

Objective: To study the anti-tumor immune effects of WT1 peptide vaccine in SCID mice with xenografted human monocytic leukemia. Methods: Twenty-four hours after intraperitoneal injection of human peripheral blood lymphocytes, the xenograft human monocytic leukemia model in SCID mice was established by subcutaneous inoculation of THP1 cells. The mice were randomly divided into three groups with eight mice in each. Blank control group was vaccinated with incomplete Freund's adjuvant (IAF). Helper T cell epitope group was vaccinated with helper T cell epitopes and IAF. WT1 group was vaccinated with WT1 peptide, helper T cell epitopes and IAF. When the tumor volume grew to 100 mm3, intraperitoneal injection of vaccine components started. The SCID mice were killed 14 days after vaccination. LDH release method was adopted to detect the specific CTL killing activity of spleen cells. Histological characteristics of tumor tissue were observed under microscope after HE staining. Flow cytometry was used to test the levels of peripheral blood CD3+/CD4+T cells, CD3+/CD8+T cells and CD4+CD25+ Treg cells. ELISA method was applied to detect the levels of serum immunoglobulin, IL-2, γ-interferon, TGF-β and IL-10. Results: The xenograft human monocytic leukemia model was successfully established in SCID mice and tumor developed in all the SCID mice. In WT1 group, the activity of mouse spleen cells on THP1 cells was significantly higher than that in helper T cell epitope group and control group (P<0.05). The mean weight and volume of tumor were significantly lower in WT1 group than in helper T cell epitope group and control group (P<0.05). A large amount of tumor cell degeneration and necrosis was observed under the microscope in WT1 group mice and few tumor cells survived. Peripheral blood levels of CD3+/CD4+T cells, CD3+/CD8+T cells, IgG, IFN-γ and IL-2 were all higher in WT1 group than in helper T cell epitope group and control group (P<0.05). However, peripheral blood levels of CD4+/CD25+Treg cells, TGF-β and IL-10 were all lower in WT1 group than in helper T cell epitope group and control group (P<0.05). Conclusion: WT1 polypeptide vaccine can effectively produce anti-tumor immunity and kill leukemia cells in SCID mice with exnografted human monocytic leukemia.

Establishment of Metrigel-VEGF-SW982 complexes and application of the complexes in synovial sarcoma model in mice / 西安交通大学学报(医学版)

Objective To establish Metrigel-VEGF-SW982 complexes and use the complexes to produce animal models of synovial sarcoma so as to provide new ideas for establishing other models of soft tissue tumors. Methods After the SW982 cells were cultured and collected,they were resuspended with Metrigel,and VEGF was added.The suspension was seeded into transwell to establish the scaffold complexes of Metrigel-VEGF-SW982.The complexes were cultured overnight.Cryosections were made and HE staining was carried out to observe the cell scaffold complexes.We randomly divided 10 female SCID mice(4 week old)into scaffold group and control group. The mice in the scaffold group were transplanted with cell-scaffold complexes,and the control group with cell suspension.After 8 weeks,the success rate of modeling was compared between two groups.The mice were sacrificed and the tumors were obtained.HE staining was carried out to observe the histopathological features of tumors in both scaffold group and control group.Results The SW982 cells were cultured with Matrigel in a 3D way,which could simulate the growth condition of cells in vivo.Establishing synovial sarcoma animal model with cell scaffold complex could increase the success rate.The tumors in scaffold group had a larger volume,higher density of tumor cells and greater vascularization(P＜0.05).Conclusion Establishing synovial sarcoma animal model with Metrigel-VEGF-SW982 complex can greatly improve the success rate of modeling,which can provide basis for the study of synovial sarcoma.

Establishment of NOD/SCID-mouse model with multiple myeloma using myeloma cells from patients / 西安交通大学学报(医学版)

Objective To explore the feasibility of establishment of NOD/SCID-mouse model with multiple myeloma by using plasma cells from myeloma patients.Methods The femurs and tibias were removed from the New Zealand white rabbits;the muscles,periosteum and cartilage tissues were cleared.Then each bone was cut into two pieces gently along its middle.The NOD/SCID mice weighing 25 - 30 g (4 - 6 weeks)were anesthetized by intraperitoneal injection;rabbit bone was inserted into the right side of the mouse back and engraftment of the bones was allowed to take place after 4 weeks.The 5000 000 purified plasma cells which expressed CD38 +/CD45 - were immunofluorescence labeled and then injected slowly into the implanted rabbit bone through the distal end.The mice were observed weekly;the plasma cells growth in mice was screened by the living-imaging system and the tumor from the mice was determined by biopsy.Results The implanted rabbit bone survived after 4 weeks.The tumor in mice was observed 2 weeks after the purified myeloma cells were injected into the rabbit bone,and it reached 100 mm3 after 8 weeks.Results of the living-imaging system showed that the myeloma cells had uptake in the rabbit bone after 2 weeks of injection and this phenomenon was more pronounced after 8 weeks of injection (2.4×10 4 vs .1.5× 10 5 ,P < 0.05 ).The tumor infiltrated with numerous plasma cells and osteoclasts increased upon the biopsy. Conclusion Rabbit bone marrow implanted into NOD/SCID mice can effectively support local injection of plasma cells of multiple myeloma patients,and the NOD/SCID-mouse model of myeloma has been established.This model can be used to study in vivo experiments related to myeloma and clinical therapeutic approaches for this disease.

The establishment of human esophageal cancer model in Hu-PBL-SCID mice / 肿瘤研究与临床

Objective To establish a mouse model for human esophageal cancer.Methods Human PBL were isolated directly from whole blood by density gradient centrifugation.Fifteen SCID mice were randomly divided into two groups.Group A was control group,and in group B there were 12 mice intraperitoneally injected with 2×107 human PBL and subcutaneously injected with 5×106 ECA109 cells.The rate of tumor transplantation,tumor growth,metastasis and histological features were observed.After 3,4,5,6 weeks of engraftment,the level of IgG in mouse serum and the spleen weight were detected.Results The successful rate of tumor transplantation was 100 ％.Metastasis was not found.After 3,4,5,6 weeks of engraftment,the spleen weight in group B were (55.44±4.45) mg,(88.62±2.24) mg,[(125.98±2.19) mg] (P ＜ 0.05) and (213.71±2.96) mg,which had statistical significance compared with the control group (41.87±2.97) mg.The level of IgG was significantly higher than that in control group (P ＜ 0.01).Conclusion The experimental results demonstrate that human esophageal cancer models have been established in Hu-PBL-SCID mice.

The pathogenesis of cartilage erosion of rheumatoid arthritis and the development of animal model by engraftment of RA synovium and normal human cartilage under the kidney capsule of the SCID mice / 中华风湿病学杂志

Objective To develop the humanized animal model for RA cartilage erosion,and study the mechanisms of its pathogenesis.Methods RA synovium and normal human cartilage under the kidney cap- sule of the SCID mice were engrafted,and were maintained for 4～16 weeks.In addition,mice underwent simi- lar surgery except the engraftment served as controls.After 4,8,12 or 16 weeks,the mice were killed and the grafts were harvested and the cartilage destruction was assessed histologically by haematoxylin/eosin-stained paraffin sections.Results Histological examination revealed the presence of infiltration of RA synovium cells into the cartilage after 4 weeks and the cartilage was destructed evidently.These studies demonstrated that the RA-SCID model maintained many of the phenotypic and functional features of RA.Conclusion This RA-SCID mouse is a useful animal model for study of the pathogenesis and the development of new drugs for RA patients.

Expression of Cell Surface Receptors on Human Glioblastoma Xenograft Model in NOD/SCID Mouse / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: To obtain basic data for development of a glioblastoma-specific immunotoxin, the expression of variable cell surface receptors on a human glioblastoma xenograft model was evaluated, using NOD/SCID mice. MATERIALS AND METHODS: We developed a xenograft model in NOD/SCID mice implanted with a human glioblastoma cell line (U-87MG). Immunohistochemical studies were performed on implanted tumor nodules (n=8) using antibodies against CD71, EGFR, IGF-IRalpha, CXCR4 and IL-4Ralpha. RESULTS: Expression of IL-4Ralpha, in implanted tumornodules, was the highest of the cell surface receptors evaluated in this study. However, the endothelial cells in, and around, the tumor nodules also revealed immunopositivity against IL-4Ralpha. The immunoreactivity of IL-4Ralpha, and other surface receptors such as CD71, IGF-IRalpha and EGFR, was prominent in tumor nodules associated with tumor necrosis. CONCLUSION: IL-4Ralpha would be a possible target for the development of glioblastoma-specific immunotoxin, although there are limitations due to its endothelial expression.

Growth and Metastasis of MCF-7 Breast Cancer Cells Grafted to huPBMC-SCID Mouse / 한국유방암학회지

PURPOSE: The severe combined immunodeficient (SCID) mice which lack the functional T and B lymphocytes have been widely used for the research of various human diseases including AIDS, transplantation, autoimmune disease and cancer. The purpose of this study was to evaluate the huPBMC-SCID mouse as an animal model for human breast cancer research. METHODS: 5x10(7) human PBMC (peripheral blood mononuclear cell) were injected intraperitoneally in 18 SCID mice. After 24 hours, 2.5x10(6), 5x10(6) and 10x10(6) MCF-7 human breast cancer cells were innoculated subcutaneously in the right flank of each of the 3 groups of 6 huPBMC-SCID mice. RESULTS: 4 subcutaneous ecchymosis (2 perioral area, 2 scalp), 1 splenomegaly and 1 hepatic embolism were found during the 20 weeks after the injections. The growth of tumor xenograft was identified in 14 of the total 18 huPBMC-SCID mice, and the growth rate of the tumor was proportional to the number of the innoculated cancer cells. Distant metastases were found in the retroperitoneum, kidney, pelvic cavity, omentum, perisplenic area and regional lymph node in 50 % of mice, but not in the lung and liver at 20 weeks. CONCLUSION: In summary, the huPBMC-SCID mouse was expected to play an important roles as an animal model of human cancers including breast cancer.

Fumagillin inhibits colorectal cancer growth and metastasis in mice:an in vivo and in vitro study of antiangiogenesis / 中国癌症杂志

Background and purpose:Fumagillin is an inhibitor of type 2 methionine aminopeptidase that can block blood vessel formation. However, its molecular mechanism and therapeutic value in colon cancer still remain to be elucidated.In this study, the effect of Fumagillin on the growth of colon cancer was examined. Methods:Twenty mice were divided into 4 groups and injected subcutaneously with 5?105/L WiDr or HT-29 cells in 200 ?L phosphate-buffered saline (PBS) respectively. After 4 weeks, intraperitoneal injections of Fumagillin (0.1 mg/kg), Cyclo (1 mg/kg), or both were given every 2 days for 4 weeks. The tumor weight and microvessel density (MVD) were examined. Geneexpression profiles were examined by microarray analysis of human umbilical endothelial cells (HUVECs). Results: The Fumagillin-treated mice showed smaller tumor mass and lower MVD-CD105 levels than control ones. In vitro proliferation and tube formation of HUVEC was also significantly decreased by Fumagillin. Microarray analysis of Fumagillin-treated HUVECs showed up-regulation of 71 genes and down-regulation of 143 genes. Expression changes were involved in cell proliferation, migration, adhesion and gene transcription. Quantitative real time-polymerase chain reaction and Western blot revealed decreased expression of cyclin E2, activated leukocyte cell adhesion molecule (ALCAM), and intercellular adhesion molecule-1 (ICAM-1) genes in the presence of Fumagillin. Conclusion: Fumagillin was found to suppress colorectal cancer growth by suppressing angiogenesis. The down-regulation of cyclin E2, ALCAM and ICAM-1 by fumagillin may be involved in the anti-angiogenesis.

Investigation of severe combined immunodeficient mice transplanted with human hematopoietic cells intraperitoneally and intravenously / 中国免疫学杂志

Objective:To compare the seeding efficiency and graft versus host disease (GVHD) of severe combined immunodeficient (SCID) mice transplanted with human hematopoietic cells intraperitoneally (ip) and intravenously (iv) and to explore the method to set up psoriatic animal model by xenogeneic bone marrow transplantation.Methods:Normal human bone marrow mononuclear cells (BMMNC) were separated by density gradient centrifugation and BMMNC (4?10~7) were injected into lethally irradiated SCID mice by ip or iv injection. GVHD symptom and periphery blood white blood cell resuming dynamics in mice were observed after xenotransplantation and flow cytometry was performed to detect human source CD45~+ cells proportion in periphery blood and bone marrow of mice.Results:The mice transplanted by tail intravenous injection presented obvious GVHD symptoms promptly 2 weeks after transplanting and only one mouse survived in 12 weeks. Among the mice received tail intravenous injection and dealed with cydosporin(CsA) and methotrexate(MTX),some of the mice showed slight GVHD symptom and survival rate was 80%(8/10) in 12 weeks. Slight GVHD symptoms appeared after human bone marrow transplantation by intraperitoneal injection and then most of mice returned to the normal and the survival rate was 70%(7/10) in 12 weeks. The periphery blood white blood cells resuming dynamics, CD45~+ cell proportion of periphery blood and bone marrow after transplantation show no significant difference between the groups transplanted by intravenous and intraperitoneal injection.Conclusion:Human hematopoietic cells could home to bone marrow in SCID mice and result in hematopoietic reconstitution. The transplantation method by intraperitoneal injection, which showed efficient seeding capability, can be used to both bone-marrow transplantation and reducing GVHD.