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Original article Inhibition of collagen-induced arthritis by dna vaccines encoding TCR Vβ5.2 and TCR Vβ8.2

Chinese Medical Journal 2009年第9期122卷 1039-1048页

作者： GE Ping-ling MA Li-ping WANG Wei LI Yun ZHAO Wen-ming Department of Transfusion Aerospace Center Hospital Beijing100049 China Department of Immunology Capital Medical University Beijing100069 China

Background Arthritogenic T lymphocytes with common T cell receptor （TCR） Vβ clonotypes, infiltrating in the articulars of rheumatoid arthritis （RA） patients, play a central role in the pathogenesis of RA. TCR Vβ5.2 and TCR Vβ8.2 are the main pathogenic T cell clonotypes in the course of collagen-induced arthritis （CIA） progression in Lewis rats. To investigate a TCR-based immunotherapy for RA, we constructed recombinant dna vaccines encoding TCR Vβ5.2 and TCR Vβ8.2, and evaluated the inhibitive effects of the two vaccines on CIA rats.Methods Genes encoding TCR Vβ5.2 and TCR Vβ8.2 were amplified by RT-PCR from spleen lymphocytes of Lewis rats and cloned into the eukaryotic expression vector pTargeT. The expression of vaccines was confirmed by RT-PCR and immunohistochemistry. The inhibitive effects of the vaccines on articulars of CIA rats were assessed with arthritis index evaluation and histology. Interferon γ （IFN-γ） and interleukin （IL）-4 production by spleen lymphocytes were tested with enzyme-linked immunospot assay （ELISPOT） technique, the changes in peripheral CD4^＋ and CD8^＋ lymphocyte populations were tested by flow cytometry, and the level of anti-CII antibody in serum was assayed by enzyme-linked immunosorbent assay （ELISA）.Results Recombinant dna vaccines pTargeT-TCR Vβ5.2 and pTargeT-pTCR Vβ8.2 were successfully constructed. Both vaccines inhibited CIA, which alleviated the arthritis index score （P 〈0.05）, decreased the level of IFN-γ （P 〈0.05）, and reduced the ratio of CD4^＋/CD8^＋ lymphocytes （P 〈0.05） and the anti-CII antibody in serum （P 〈0.05）. In addition, the histological change in dna-vaccinated rats was less serious than CIA rats. Compared to pTCR Vβ 8.2 and pTCR Vβ 5.2 groups, the group that was injected with a combination of the two vaccines showed stronger inhibitive effects on CIA than either individual vaccine.Conclusion The recombinant plasmids pTargeT-TCR Vβ5.2 and pTargeT-TCR Vβ8.2 have obvious inhibatory effects on CIA rats and bet

关键词： arthritis, collagen-induced T cell receptor vaccines, dna

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Hepatitis E virus chimeric dna vaccine elicits immunologic response in mice

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World Journal of Gastroenterology 2005年第42期11卷 6713-6715页

作者： Yan Hong Bing Ruan Lian-Hua Yang Yong Chen Luo Jing Yi-Ting Wang Hua-Jun Hu Institute of Bioengineering.ZhejiangAcademy of Medical Sciences Hangzhou 310013 ZhejiangProvince China Zhejiang University Medical School Hangzhou310003 Zhejiang Province China

AIM： To construct the plasmid pcHEV23 containing fragments of HEV ORF2 and ORF3 chimeric gene and to assess its ability to elicit specific immunologic response in mice. METHODS： The gene encoding the structural protein of HEV ORF2 fragment and full-length ORF3 was amplified by PCR. The PCR products were cloned into an eucaryotic expression plasmid pcdna3. The resulting plasmid pcHEV23 was used as a dna vaccine to inoculate BALB/c mice intramuscularly thrice at a dose of 100 or 200 ug. Mice injected with empty pcdna3 dna or saline served as control and then specific immune responses in the mice were detected. RESULTS： After 2-3 times of inoculation, all mice injected with pcHEV23 had anti-HEV IgG seroconversion and specific T lymphocyte proliferation. The lymphocyte stimulation index in the group immunized with pcHEV23 （3.1＋0.49） was higher than that in the control group （0.787±0.12, P〈0.01）. None in the control group had a detectable level of anti-HEV IgG. CONCLUSION： dna vaccine containing HEV ORF2 and ORF3 chimeric gene can successfully induce specific humoral and cellular immune response in mice.

关键词： Hepatitis E virus Animals Female Humans Lymphocyte Activation Mice Mice, Inbred BALB C Open Reading Frames Plasmids Recombinant Fusion Proteins Research Support, Non-U.S. Gov't T-Lymphocytes vaccines, dna Viral Hepatitis vaccines

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Protective effect of dna-mediated immunization with a combination of SAG1 and IL-2 gene adjuvant against infection of Toxoplasma gondii in mice

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Chinese Medical Journal 2002年第10期115卷 1448-1452页

作者：陈观今陈海峰郭虹郑焕钦汪琦中山大学中山医学院寄生虫学研究所广州510089

OBJECTIVE: To characterize the immune response induced by SAG1 encoding plasmid combined with IL-2 gene adjuvant in mice and to assess the protective effect of this vaccination against toxoplasmosis. METHODS: Mice were co-injected intramuscularly with plasmid encoding Toxoplasma gondii SAG1 plus murine IL-2 expression vector at a dose of 100 microg. Booster immunizations were employed 2 more times at 3-week interval. As controls, mice were inoculated with PBS or empty plasmid pcdna3. Humoral and cellular responses were assayed using ELISA for the determination of Ab, Ab isotype and IFN-gamma, as well as IL-4. To detect the integration and dissemination of dna in the injected mice, PCR and in situ hybridization were performed. All mice were then infected with highly virulent RH tachyzoites of Toxoplasma gondii intraperitoneally. RESULTS: Significant increases in specific IgG levels were observed in mice after immunization three times with SAG1 expression plasmid. With respect to the IgG isotype, co-inoculation of IL-2 expression plasmid enhanced the level of IgG2a and the production of IFN-gamma. Challenging mice by vaccinating with combined plasmids with RH tachyzoites resulted in prolonged survival. CONCLUSION: Humoral and cytokine responses elicited by SAG1 dna immunization can be modulated by co-inoculation with IL-2 expression plasmid. The use of dna vaccine in combination with an appropriate cytokine gene to prevent T. gondii infection warrants further investigation.

关键词： Antigens, Protozoan Animals Antibodies, Protozoan Cytokines Female Immunization Immunoglobulin G Interleukin-2 Mice Protozoan Proteins Protozoan vaccines Research Support, Non-U.S. Gov't Toxoplasma Toxoplasmosis, Animal vaccines, dna

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Humoral and cellular immunogenecity of dna vaccine based on hepatitis B core gene in rhesus monkeys

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World Journal of Gastroenterology 2001年第1期7卷 102-106页

作者： Zu Hu Huang1 Hui Zhuang2 Shan Lu3 Ren Hua Guo1 Guo Min Xu2 Jie Cai1 Wan Fu Zhu2 1Department of Infectious Diseases. The First Affiliated Hospital of Nanjing Medical University, Nenjing 210029, Jiangsu Province. China2Faculty of Microbiology, Beijing University, Beijing 100000, China3University of Massachusetts Medical Center The First Affiliated Hospital of Nanjing Medical University 300 Guangzhou Road Nanjing 210029 Jiangsu Province China.

INTRODUCTIONHepatitis B virus (HBV) is the most commonetiologic agent for infectious liver diseases. It isestimated that there are more than 250 millionchronic HBV carriersin the world today and thereis a significant ... 详细信息

关键词： vaccines, dna Animals Antibodies, Viral Antibody Formation Antibody Specificity Cell Division Cells, Cultured Enzyme-Linked Immunosorbent Assay Female Hepatitis B control Hepatitis B Core Antigens Immunity, Cellular Immunoglobulin G Interferon Type II Interleukin-4 Leukocytes, Mononuclear Macaca mulatta Male Research Support, Non-U.S. Gov't

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pcdnaL1 genetic immunization can induce specific cell-mediated immune responses in C57BL/6 mice

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Chinese Medical Journal 2002年第11期115卷 1697-1700,154页

作者：宋建明刘天菊孙向乐王一理司履生深圳市第二人民医院病理科深圳518035 西安交通大学生命科学与技术学院肿瘤研究所西安710061

OBJECTIVE: To investigate the specific cell-mediated immune efficacy of the an HPV16 prophylactic vaccine. METHODS: C57BL/6 mice were randomly divided into 3 groups: experimental group I (treated with pcdna L1), control group II (treated with pcdna3.1) and control group III (treated with PBS buffer). The mice were immunized three times during a three-week interval. Ten to fourteen days after the third inoculation, a footpad swelling test was used to detect delayed-type hypersensitivity (DTH) responses. Antigen-specific splenocyte proliferation assay and quantitation of IFN-gamma cells in splenocytes were performed by FACS assay. RESULTS: In the experimental group, splenocytes actively proliferated after stimulation with HPV16 VLP, and had developed a markedly larger amount of CD8(+) IFN-gamma(+) cells, which is an index for special CTL. Also, the footpad was significantly thickened upon inoculation with HPV16 VLP. CONCLUSION: Naked dna vaccine of HPV16 L1 can induce specific cell-mediated immune responses in mice, which should be considered for evaluation of HPV16 dna vaccine feasibility.

关键词： Animals Hypersensitivity, Delayed Immunization Interferon Type II Lymphocyte Activation Mice Mice, Inbred C57BL Papillomavirus, Human Research Support, Non-U.S. Gov't Spleen vaccines, dna Viral vaccines

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