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Construction and humoral immune response of Epstein-Barrvirus latent membrane protein 2 DNA vaccine in mice

Frontiers of Medicine 2009年第4期3卷 390-395页

作者： Jianqing PAN Qin ZHANG Daowen WANG Department of Internal Medicine Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China Research Center for Neural Engineering Shenzhen Institute of Advanced TechnologyChinese Academy of ScienceShenzhen 518055China

We constructed a eukaryotic expression plas-mid encoding Epstein-Barr virus latent membrane protein 2(EBV,LMP2)and evaluated its effects on humoral ***,the encoding sequence of the EBV LMP2 was ampliﬁed from B95-8 cell RNA by reverse transcrip-tion polymerase chain reaction(RT-PCR)and then was directionally cloned into eukaryotic expression vector *** was employed to evaluate immune response of the mice inoculated doubly with the DNA *** serum antibody against LMP2 was detected with enzyme-linked immunosorbent assay(ELISA).The recombinant plasmid pcDNA3.1-LMP2 was conﬁrmed by the restrictive endonuclease analysis and sequence *** serum titer of IgG antibody against LMP2 epitope in the mice immunized with the DNA vaccine encoding LMP2 was up to 1∶*** conclusion,the EBV LMP2 DNA vaccine can induce a strong humoral immune response in mice.

关键词： Epstein-Barr virus latent membrane protein 2 nasopharyngeal carcinoma humoral immunity

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Analysis of Epstein-Barr viral DNA load, EBV-LMP2 specific cytotoxic T-lymphocytes and levels of CD4^＋CD25^＋ T ceils in patients with nasopharyngeal carcinomas positive for IgA antibody to EBV viral capsid antigen

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Chinese Medical Journal 2009年第10期122卷 1173-1178页

作者： MO Wu-ning TANG An-zhou ZHOU Ling HUANG Guang-wu WANG Zhan ZENG Yi Laboratory Department First Affiliated Hospital Guangxi Medical University Nanning Guangxi 530021 China Department of Otolaryngology & Head and Neck Surgery First Affiliated Hospital Guangxi Medical University Nanning Guangxi 530021 China State Key Laboratory for Infectious Disease Prevention and Control Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention Beijing 100052 China

Background Epstein-Barr virus （EBV） is a herpesvirus commonly associated with several malignant diseases including nasopharyngeal carcinoma （NPC）, which is a common cancer in Southeastern Asia. Previous studies showed that plasma levels of EBV-DNA might be a sensitive and reliable biomarker for the diagnosis, staging and evaluating of therapy for NPC. There are a few analyses of the levels of EBV-latent membrane protein 2 （LMP2）-specific cytotoxic T-lymphocytes （CTLs） in patients with NPC. This study was conducted to investigate the levels of EBV-LMP2-specific CTLs, EBV-DNA load and the level of CD4^＋CD25^＋T cells in such patients. Methods From February 2006 to April 2006, 62 patients with NPC, 40 healthy virus carriers positive for EBV viral capsid antigen （EBV-IgA-VCA） and 40 controls were enrolled in the study. We used a highly sensitive ELISPOT assay, real-time polymerase chain reaction （PCR） and flow cytometry to measure the EBV-LMP2-specific CTL response, the EBV DNA load and the level of CD4^＋CD25^＋T cells, respectively. Results The EBV-LMP2-specific CTL responses of the samples from the control, healthy virus carriers and patients with NPC were significantly different from the LMP2 epitopes, with the control and healthy virus carrier samples displaying a stronger response in three cases. There were significant differences in EBV DNA load in serum between NPC and the healthy groups; patients with NPC at stages Ⅲ or Ⅳ had significantly higher viral loads compared with those at stages Ⅰ or Ⅱ. A significantly higher percentage of CD4^＋CD25^＋ T lymphocytes were detected in the patients, compared with healthy virus carriers and healthy controls. Moreover, patients with advanced stages of NPC （Ⅲ and Ⅳ） had significantly higher percentages than the patients with early stages （Ⅰ and Ⅱ）. Conclusions Patients with NPC are frequently unable to establish or maintain sufficient immunosurveillance to control proliferating B cells harboring EBV and to destroy the tumor

关键词： nasopharyngeal carcinoma cell immunity Epstein-Barr virus latent membrane protein 2 cytotoxic T lymphocyte

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Constructing TC-1-CLUC-LMP2 Model Tumor Cell to Evaluate the Anti-Tumor Effects of LMP2-Related Vaccines

Constructing TC-1-CLUC-LMP2 Model Tumor Cell to Evaluate the...

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第十四届全国免疫学学术大会

作者： Liying Sun Yanzhe Hao Zhan Wang Yi Zeng 中国疾病预防控制中心病毒病预防控制所

Epstein-Barr virus(EBV) is related to a variety of malignant tumors,and its encoded protein,latent membrane protein 2(LMP2),is an effective to rget antigen that is widely used to construct vector ***,the model cells carrying LMP2 have still not been established to assess the oncolytic effect of LMP2-related vaccines at *** this study,TC-1-GLUC-LMP2 tumor cells were constructed as target cells to evaluate the anti-tumor effects of LMP2-assosiated *** results showed that both LMP2 and Gaussia luciferase(GLuc) genes could be detected by polymerase chain reaction(PCR) and reverse transcription-polymerase chain reaction(RT-PCR) in TC-1-GLUC-LMP2 *** blot results showed that the LMP2 and Gaussia luciferase proteins were stably expressed in tumor cells for at least 30 *** mixed 5 × 10 LMP2-specific mouse splenic lymphocytes with 5 × 10 TC-1-GLUC-LMP2 target cells and found that the to rget cells were killed as the specific killing effect was obviously enhanced by the increased quantities of LMP2-peptide stimulated ***,the tumor cells could not be observed in the mice inoculated TC-1-GLUC-LMP2 cells after being immunized with vaccine-LMP2,while the vaccine-NULL immunized mice showed that tumor volume gradually grew with increased inoculation *** results indicated that the TC-1-GLUC-LMP2 cells stably expressing LMP2 and GLuc produced tumors in mice,and that the LMP2-specific cytotoxic T lymphocyte(CTL) effectively killed the cells in vitro and in vivo,suggesting that TC-1-GLUC-LMP2 cells can be used as model cells to assess the immune and antitumor effects of LMP2-related vaccines.

关键词： Epstein-Barr virus latent membrane protein 2 Gaussia luciferase cytotoxic T lymphocyte in vivo imaging system

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