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CONSTRUCTION OF HU-PBL/scid CHIMERAS AND DEVELOPMENTOF EBV-RELATED LYMPHOMAS

Chinese Medical Sciences Journal 2005年第1期20卷 16-22,页

作者： Run-liangGan KeLan Zhi-huaYin Li-jiangWang YingSong Kai-taiYao CancerResearchInstitute SchoolofMedicineNanhuaUniversityHengyang421001 KeyLabofCarcinogenesisofChineseHealthMinistry XiangyaMedicalCollegeCentralSouthUniversityChangsha410078

Objective To construct hu-PBL/scid chimeras and to investigate the development of lymphoma and oncogenicity of the Epstein-Barr virus (EBV). Methods Human peripheral blood lymphocytes (PBLs) were isolated from healthy adult donors and transplanted intraperitoneally into severe combined immunodeficient(scid) mice. Mice with hu-PBL engraftment from healthy EBV seronegative donors were injected intraperitoneally with EBV-containing supernatant from suspension culture of B95-8 cell line (active infection), whereas mice receiving lymphocytes from healthy EBV seropositive donors were not re-infected with B95-8 derived EBV (latent infection). Pathological examination and molecular analysis were performed on experime-ntal animals and induced neoplasms. Results In the early stage of this experiment, 12 mice died of acute graft-versus-host disease, mortality was 34.3% (12/35 mice) with an average life span of 17.5 days. In 19 survival hu-PBL/scid chimeric recipients from 12 healthy donors, tumor incidence was 84.2% (16/19 mice). The average survival time of tumor-bearing mice was 65.5 days. EBV-related neoplasms in scid mice were nodular tumors with aggressive and fatal features. Histological morphology of tumors exhibited diffuse large cell lymphomas. Immunohistochemistry revealed that LCA (CD45) and L26 (CD20) were positive, but both PS1 (CD3) and UCHL-1 (CD45RO) were negative, and EBV products ZEBRA, LMP1, and EBNA2 were expressed in a small number of tumor cells. EB virus particles were seen in the nuclei of some tumor cells by electron microscopy, and EBV DNA could be amplified in the tumor tissues by PCR. In situ hybridization indicated that the nuclei of tumor cells contained human-specific Alu sequence. Conclusions EBV-induced tumors were human B-cell malignant lymphomas. We obtained direct causative evidence dealing with EBV-associated tumor deriving from normal human cells.

关键词： lymphoma Epstein-Barr virus oncogenicity scid mouse

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Hepatocellular carcinoma xenograft supports HCV replication:A mouse model for evaluating antivirals

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World Journal of Gastroenterology 2011年第3期17卷 300-312页

作者： Sidhartha Hazari Henry J Hefler Partha K Chandra Bret Poat Feyza Gunduz Tara Ooms Tong Wu Luis A Balart Srikanta Dash Department of Pathology and Laboratory Medicine Tulane University Health Sciences Center New Orleans LA 70112 United States Department of Gastroenterology and Hepatology Tulane University Health Sciences Center New Orleans LA 70112 United States Department of Comparative Medicine Tulane University Health Sciences Center New Orleans LA 70112 United States

AIM: To develop a hepatocellular carcinoma (HCC) xenograft model for studying hepatitis C virus (HCV) replication in a mice, and antiviral ***: We developed a stable S3-green fluorescence protein (GFP) cell line that replicated the GFP-tagged HCV sub-genomic RNA derived from a highly efficient JFH1 virus. S3-GFP replicon cell line was injected subcutaneously into γ-irradiated scid mice. We showed that the S3-GFP replicon cell line formed human HCC xenografts in scid mice. Cells were isolated from subcutaneous tumors and then serially passaged multiple times in scid mice by culturing in growth medium supplemented with G-418. The mouse-adapted S3-GFP replicon cells were implanted subcutaneously and also into the liver of scid mice via intrasplenic infusion to study the replication of HCV in the HCC xenografts. The tumor model was validated for antiviral testing after intraperitoneal injection of interferon-α (IFN-α). RESULTS: A highly tumorigenic S3-GFP replicon cell line was developed that formed subcutaneous tumors within 2 wk and diffuse liver metastasis within 4 wk in scid mice. Replication of HCV in the subcutaneous and liver tumors was confirmed by cell colony assay, detection of the viral RNA by ribonuclease protection assay and real-time quantitative reverse transcription polymerase chain reaction. High-level replication of HCV sub-genomic RNA in the tumor could be visualized by GFP expression using fluorescence microscopy. IFN-α cleared HCV RNA replication in the subcutaneous tumors within 2 wk and 4 wk in the liver tumor model. CONCLUSION: A non-infectious mouse model allows us to study replication of HCV in subcutaneous and metastatic liver tumors. Clearance of HCV by IFN-α supports use of this model to test other anti-HCV drugs.

关键词： Hepatitis C virus Hepatocellular carcinoma Tumor xenograft scid mouse Interferon-α Antiviral agent Virus replication

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Effects of dendritic cell vaccines on hematogenous micrometastasis of bladder cancer carrying for PBL-scid mice

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The Chinese-German Journal of Clinical Oncology 2009年第6期8卷 341-345页

作者： Bin Wang Zhenguo Mi Zhibin Li Xinjing Yang Jianwu Liu Jiwen Song Huiqing Chen Department of Urology Shanxi Tumor Hospital Taiyuan 030013 China

Objective： To study the effect of dendritic cells loaded with whole tumor antigen on hematogenous micrometastasis of bladder cancer model in hu-PBL-scid mice. Methods： T24-3 ceil subset was selected from human bladder transitional cell carcinoma T24 cell line by Boyden chamber system. The scid mice intraperitoneally injected with 4 × 10^7 hu-PBL and subcutaneously injected with 3 × 10^6 T24-3 cells were named hu-PBL-T24-3-scid model. Human IgG level in the blood plasma of mice was detected by ELISA, and human CD3^＋, CD4^＋, CD8^＋ T cells in blood and spleen cells of mice were detected by FCM analysis for human immune reconstruction study. Human CK20 mRNA expression in mice peripheral blood was detected by RT-PCR to investigate metastasis of tumor cells. The PBMCs were isolated from human peripheral blood, and were induced into DCs by co-culture with rhGM-CSF and rhlL-4 in vitro. The DC vaccines were produced by co-culturing with whole tumor antigen which was purified through freezing and melting T24-3 cell subset. After T24-3 cells injected into scid mice for 5 weeks, the mice were treated with DC vaccines. Results： All mice were initially treated at 5th week. The expression of CK20 mRNA in peripheral blood of DC vaccines treated mice was the lowest. There was 2 mice showing CK20 mRNA expression and 3 mice with metastasis tumor in PBS group. MMP-7 mRNA expression in tumor tissues of DC vaccines treated mice was statistically lower than that of PBS group （P 〈 0.01）. Conclusion： DC vaccines have a good effect on hu-PBL-scid mice bladder cancer model by reducing hematogenous micrometastasis.

关键词： dendritic cells bladder tumor scid mouse immune reconstruction rnicrometastasis

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Comparative serum metabolomics between scid mice and BALB/c mice with and without Schistosoma japonicum infection:insights into the abnormal growth and development of schistosome in scid mice

Comparative serum metabolomics between SCID mice and BALB/c ...

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第十一届全国寄生虫学青年工作者学术研讨会

作者： Rong Liu Feng Ye Qinping Zhong Shuhong Wang Ting Chai Huifen Dong Zhenping Ming School of Basic Medical Sciences Wuhan University

Objective The small blood flukes of genus Schistosoma,which cause one of the most prevalent and serious parasitic zoonosis schistosomiasis,are dependent on immunerelatedfactors from their mammalian host to facilitate their growth and development,and granulomatous pathology caused by eggs deposited in host’s liver and intestinal *** development is hampered in the mice lacking just T cells,and is even worse in the severe combined immunodeficient(scid) mice lacking both T and B ***,it’s still not clear about the underlying regulatory mechanisms of schistosome growth and development by host’s immune *** This study,therefore,detected and compared the serum metabolic profiles between the immunodeficient scid mice and immunocompetent BALB/c mice before and after *** infection for thirty-five days using liquid chromatography-mass spectrometry(LC-MS).Results Totally,705 ion features in electrospray ionization in positive-ion mode(ESI+) and 242 ion features in ESI-mode wereidentified,*** Component Analysis(PCA) and Orthogonal Partial Least Squares Discriminant Analysis(OPLS-DA) identified several metabolite sets that clearly differentiated samples of scid mice from those of BALB/c mice before and after *** ***,distinct serum metabolic profiles were identified between scid mice and BALB/c mice without ***,common and uniquely perturbed serum metabolites and their involved pathwayswere identified between scid mice and BALB/c mice with *** *** results revealedthe involvedsphingolipid metabolism,glycerophospholipid metabolism,glycosylphosphatidylinositol(GPI)-anchor biosynthesis,arachidonic acid metabolism,linoleic acid metabolism and alpha-linolenic acid metabolism were associated withthe retarded growth and development of *** in scid *** This research provides a new insight into the underlying molecular mechanisms of host’s systemic metabolic

关键词： Schistosoma japonicum scid mouse BALB/c mouse growth and development LC-MS/MS serum metabolomics

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