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dna vaccine encoding Der p2 allergen down-regulates STAT6 expression in mouse model of allergen-induced allergic airway inflammation

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Chinese Medical Journal 2006年第3期119卷 185-190页

作者： QIU Jing LI Guo-ping LIU Zhi-gang RAN Pei-xing ZHONG Nan-shan College of Life Sciences Shenzhen University Shenzhen 518060China Affiliated Hospital Luzhou Medical College Guangzhou Institute of Respiratory Diseases Guangzhou MedicalCollege Guangzhou 510120 China

Background Activation of signal transducer and activator of transcription 6 （STAT6） plays a critical role in the late phase of Th2-dependent allergy induction. STAT6 is essential to Th2 cell differentiation, recruitment, and effector function. Our previous study confirmed that dna vaccination inhibited STAT6 expression of spleen cells induced by allergen. In the present study, we determined whether dna vaccine encoding Dermatophagoides pteronyssinus group 2 （Der p2 ） could down-regulate the expression and activation of STAT6 in lung tissue from asthmatic mice. Methods After dna vaccine immunization, BALB/c mice were sensitized by intmperitoneal injection and challenged by intranasal instillation of rDer p2. The levels of the cytokines IL-4 and IL-13 in BAL fluid were measured by enzyme-linked immunosorbent assay. The lung tissue was assessed by immunohistochemical staining with anti-STAT6. The protein expression of STAT6 was determined by Western blot. The activation of STAT6 binding ability was analyzed with electrophoretic mobility shift assay. Results dna vaccine encoding Der p2 allergen effectively decreased the levels of IL-4 and IL-13 in the asthmatic mice. Histological evidence and Western blot showed that the expression of STAT6 in the dna treated mice was markedly attenuated. STAT6 binding to specific dna motif in lung tissue from the gene vaccinated mice was inhibited. Condusion dna vaccine encoding Der p2 prevents allergic pulmonary inflammation probably by inhibiting the STAT6 signaling pathway in mice with Der p2 allergen-induced allergic airway infammafion.

关键词： dna vaccine Dermatophagoides pteronyssinus group 2 allergic airway inflammation signal transducer and activator of transcription 6 （STAT6）

dna vaccine encoding L7/L12-P39 of Brucella abortus induces protective immunity in BALB/c mice

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Chinese Medical Journal 2006年第4期119卷 331-334页

作者： LUO De-yan LI Peng XlNG Li ZHAO Guang-yu SHI Wei ZHANG Song-le WANG Xi-liang Institute of Microbioiogy and Epidemiology Academy of Military Medical Sciences Beijing 100071 China

Brucella abortus is a gram-negative, facultative, intracellular bacterium that infects both cattle and humans, causing abortion and infertility in the former and undulant fever, endocarditis, arthritis, and osteomyelitis. Resistance to Brucella depends on acquired cell-mediated immunity （CMI）. Live attenuated vaccines can stimulate strong CMI response, which are usually very effective against brucellosis and are used to control brucellosis in domestic animals. However, there is no safe and effective vaccine available for human because the vaccine strains used for animals are considered too virulent for humans. A vaccine that will be noninfectious to humans but effective in stimulating a broad protective immune response is needed.

关键词： Brucella abortus dna vaccine fusion protein

Vaccination against Schistosoma japonicum Infection by dna vaccine Encoding Sj22.7 Antigen

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Acta Biochimica et Biophysica Sinica 2007年第1期39卷 27-36页

作者： Gan DAI Shiping WANG Junlong YU Shaorui XU Xianchu PENG Zhuo HE Xueqin LIU Songhua ZHOU and Fen LIU Institute of Schistosomiasis Research.Xiangya School of Medicine,Central South University,Changsha 410078 China Institute of Schistosomiasis Research Xiangya School of Medicine Central South UniversityChangsha 410078 China

To observe the in vitro expression of dna vaccine pcdna3-Sj22.7 and its immunologicaleffect in mice,the recombinant plasmid pcdna3-Sj22.7 was used to transfect HeLa cells with liposome-mediated method and the expression of Sj22.7 mRNA and protein was examined using reversetranscription-polymerase chain reaction, sodium dodecylsulfate-polyacrylamide gel electrophoresis andWestern blot.Then,the ability of pcdna3-Sj22.7 to protect against Schistosoma japonicum challengeinfections was analyzed according to worm reduction rate and egg reduction rate after vaccination of mice.The serum levels of specific IgG antibody and T lymphocyte proliferation response were also determined.After the challenge infection,Sj22.7-driven interferon (IFN)-γand interleukin (IL)-4 was also quantified.Results showed that pcdna3-Sj22.7 could express Sj22.7 mRNA and protein in vitro.Immunizationresulted in a worm reduction rate of 29.70%,egg reduction rate of 47.25% (liver) and 51.73% (intestine),and egg reduction rate of 25.90% (eggs per female),suggesting induction of significant anti-fecundityin the pcdna3-Sj22.7 group. Enzyme-linked immunosorbent assay and Western blot analysis indicatedthat immunized mice generated specific IgG against Sj22.7.T lymphocytes from mice immunized withpcdna3-Sj22.7 showed a significant proliferation response to rSj22.7.The culture of spleen cells showedthat secretion of IFN-γ increased but IL-4 decreased. The results indicate that dna vaccination bypcdna3-Sj22.7 is sufficient to elicit significant levels of protective immunity against S.japonicum infection.The dna vaccine could induce significant cellular and humoral immune response,and display predominantT helper cell type 1 type immune responses,which contribute to the protective immunity against challengeinfection in mice.

关键词： response Schistosoma japonicum dna vaccine Sj22.7 gene expression immune protection immune

dna vaccine pCD-Sj32 and its efficacy of protective immunity against infection of Schistosoma japonicum

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Chinese Medical Journal 1999年第8期112卷 5-8页

作者：石佑恩谢来平李传明韩家俊 Department of Parasitology Tongji Medical University Wuhan 430030 China Department of Immunology University of Alabama Briminghan al(USA)

Objective To study protective immunity afforded by murine immunization with dna vaccine of Schistosoma japonicum (S. japonicum) as measured by reduction in worm burden and host antibody, cytokines.Methods dna vaccine pCD Sj32 was constructed. identificated and expressed. pCD Sj32 could induce substantial protective immunity against infection of S. japonicum in BALB/c mice. The best efficacy can be produced with one injection of 100?μg dna into the quadriceps muscle, combined with challenge for 8 weeks after immunization. T lymphocyte subsets of CD 8 +, IL 2. TNF and IFN γ of experimental animal could play important roles in regulating immune functions of schistosomiasis. Results High titre of specific antibody IgG could be induced by vaccinated with pCD Sj32, and antibody can mediate macrophage to produce ADCC effects in vitro. Conclusion pCD Sj32 may represent a new approach to developing subunit vaccine.

关键词： Schistosoma japonicum dna vaccine protective immunity

Protection of Guinea Pigs against Leptospira interrogans Serovar Lai by LipL21 dna vaccine

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Cellular & Molecular Immunology 2008年第5期5卷 385-391页

作者： Hanjiang He Wenyu Wang ZhongdaoWu Zhiyue Lv Jun Li Lizhi Tan Department of Parasitology Zhongshan Medical College Sun Yat-sen University Guangzhou 510080 China Key Laboratory for Tropical Disease Control Chinese Ministry of Education Guangzhou 510080 China Department of Pathogenic Biology Xiangnan University Chenzhou 423000 China Institute of Pathogenic Biology Nanhua University Hengyang 421001 China These authors contributed equally to this work

In this study, the full lipL21 gene fragment encoding outer membrane protein LipL21 was cloned from L. interrogans serovar Lai and inserted into eukaryotic expression vector pcdna3.1（＋）. The guinea pigs were immunized with pcdna3.1（＋）-lipL21, pcdna3.1（＋） or PBS. Six weeks after the second immunization, the splenocytes were isolated to detect their proliferative ability by lymphocyte transformation experiments. In addition, microscopic agglutination test was used for quantitative detection of specific antibodies. The rest guinea pigs were challenged intraperitoneally with L. interogans sorevar Lai. Then, protective effect was evaluated on the basis of survival and histopathological lesions in the kidneys, lungs, and liver. The lipL21 gene was successfully expressed in COS-7 cells through recombinant pcdna3.1（＋）-lipL21. The titer of specific antibodies substantially increased, and the stimulation index of splenocytes increased significantly. Hence, the pcdna3.1（＋）-lipL21 could protect the immunized guinea pigs from homotypic Leptospira infection. Furthermore, no obvious pathologic changes were observed in the pcdna3.1（＋）-lipL21 immunized guinea pigs. The results showed that the protective effect with pathogenic strains of Leptospira was shared by LipL21 mediated through a plasmid vector. Consequently, these results indicated that the lipL21 dna vaccine was a promising candidate for the prevention of leptospirosis.

关键词： Leptospira lipL21 dna vaccine immunoprotection

Therapeutic Effects of dna vaccine on Allergen-Induced Allergic Airway Inflammation in Mouse Model

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Cellular & Molecular Immunology 2006年第5期3卷 379-384页

作者： Guoping Li Zhigang Liu Nanshan Zhong Bin Liao Ying Xiong Inflammafion ＆ Allergic Diseases Research Unit Affiliated Hospital of Luzhou Medical College Luzhou 646000 Sichuan China College of Life Sciences Shenzhen University Shenzhen 518060 China Guangzhou Institute of Respiratory Diseases Guangzhou Medical College Guangzhou 510120 China Department of Respiratory Disease Affiliated Hospital of Luzhou Medical College Luzhou 646000 China

Vaccination with dna encoding Dermatophagoides pteronyssinus group 2 （Der p 2） allergen previously showed its effects of immunologic protection on Der p 2 allergen-induced allergic airway inflammation in mice. In present study, we investigated whether dna vaccine encoding Der p 2 could exert therapeutic role on allergen-induced allergic airway inflammation in mouse model and explored the mechanism of dna vaccination in asthma specific-allergen immunotherapy. After sensitized and challenged by Der p 2, the BALB/c mice were immunized with dna vaccine. The degrees of cellular infiltration were scored. IgE levels in serum and IL-4/IL-13 levels in BALF were determined by ELISA. The lung tissues were assessed by histological examinations. Expressions of STAT6 and NF-kB in lung were determined by immunohistochemistry staining. Vaccination of mice with dna vaccine inhibited the development of airway inflammation and the production of mucin induced by allergen, and reduced the level of Der p 2-specific IgE level. Significant reductions of eosinophil infiltration and levels of IL-4 and IL-13 in BALF were observed after vaccination. Further more, dna vaccination inhibited STAT6 and NF-kB expression in lung tissue in Der p 2-immunized mice. These results indicated that dna vaccine encoding Der p 2 allergen could be used for therapy of allergen-induced allergic airway inflammation in our mouse model. Cellular ＆ Molecular Immunology.

关键词： dna vaccine Dermatophagoides pteronyssinus group 2 allergic airway inflammation

A new dna vaccine fused with the C3d-p28 induces a Th2 immune response against amyloid-beta

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Neural Regeneration Research 2013年第27期8卷 2581-2590页

作者： Wanshu Guo Sha Sha Tongzi Jiang Xiaona Xing Yunpeng Cao Department of Neurology First Affiliated Hospital of China Medical University Department of Neurology First People's Hospital of Shenyang City

To enhance anti-amyloid-beta （Aβ） antibody generation and induce a Th2 immune response, we constructed a new dna vaccine p（Aβ3-10 ）10-C3d-p28.3 encoding ten repeats of Aβ3-10 and three copies of C3d-p28 as a molecular adjuvant. In this study, we administered this adjuvant intramus-cularly to female C57BL/6J mice at 8-10 weeks of age. Enzyme linked immunosorbent assay was used to detect the titer of serum anti-Aβ antibody, isotypes, and cytokines in splenic T cel s. A 3-（4,5-cimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide assay was used to detect the prolifera-tion rate of splenic T cel s. Brain sections from a 12-month-old APP/PS1 transgenic mouse were used for detecting the binding capacities of anti-Aβ antibodies to Aβ plaques. The p（Aβ3-10）10-C3d-p28.3 vaccine induced high titers of anti-amyloid-βantibodies, which bound to Aβplaques in APP/PS1 transgenic mouse brain tissue, demonstrating that the vaccine is effective against plaques in a mouse model of Alzheimer’s disease. Moreover, the vaccine elicited a pre-dominantly IgG1 humoral response and low levels of interferon-γ in ex vivo cultured splenocytes, indicating that the vaccine could shift the cel ular immune response towards a Th2 phenotype. This indicated that the vaccine did not elicit a detrimental immune response and had a favorable safety profile. Our results indicate that the p（Aβ3-10）10-C3d-p28.3 vaccine is a promising immunothera-peutic option for Aβvaccination in Alzheimer’s disease.

关键词： neural regeneration Alzheimer＇s disease amyloid-β C57BL/6J mice dna vaccine activeimmunotherapy passive immunotherapy C3d-p28 molecular adjuvant Th2 immune response grants-supported paper neuroregeneration

Construction of sperm-specific lactate dehydrogenase dna vaccine and experimental study of its immuno-contraceptive effect on mice

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Science China(Life Sciences) 2008年第4期51卷 308-316页

作者： CHEN Yong ZHANG Duo XIN Na XIONG YongZhong CHEN Ping LI Bo TU XiangDong LAN FengHua Research Laboratory of Medical Genetics PLA Institute of Technology for Birth ControlFetal Health and Child GrowthFuzhou General Hospital of Nanjing Military RegionFuzhou 350025China

Lactate dehydrogenase C4 (LDHC4) is a key enzyme for sperm metabolism. It is distributed specifically in testis and is highly immunogenic. In this study, two dna vaccines pVAX1-hLDHC and pVAX1-mLDHC were constructed by inserting coding sequences of human and mice LDHC4 into the eukaryotic ex-pression vector pVAX1. The production of LDHC4 specific antibodies was induced in the sera of vac-cinated mice and the reproductive tract secretions of vaccinated female mice through immunization by mucosal surface instillation. Furthermore, the antibody titer increased with the times of immunization. In the mating experiment, the number of newborns of the vaccinated mice reduced significantly and some immunized female mice even lost the ability to bear any offsprings, suggesting that the difference between the immunized and control mice was statistically significant. Sperm agglutination analysis indicated that both the antisera from immunized mice and the reproductive tract secretions of vacci-nated female mice could agglutinate normal sperms. Results of immunohistochemistry showed that the antibodies present in the sera of immunized mice and the reproductive tract secretions of vaccinated female mice could specifically react with LDHC4 antigen, which mainly locates in the cytoplasm, acrosome membrane externa and acrosome capsule of the sperm. Taken together, our results indicated that the constructed contraceptive dna vaccines did yield immunocontraceptive effects on mice and this would enable clinical trials in near future.

关键词： sperm-specific lactate dehydrogenase dna vaccine contraceptive vaccine mucosal immunization

Immunogenicity and protective efficacy of dna vaccine against visceral leishmaniasis in BALB/c mice

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The Journal of Biomedical Research 2016年第4期30卷 304-313页

作者： Sukhbir Kaur Tejinder Kaur Jyoti Joshi Parasitology Laboratory Department of Zoology Panjab University Chandigarh 160014 India

The current study was designed to examine the protective efficacy of dna vaccines based on gp63 and Hsp70 against murine visceral leishmaniasis. Inbred BALB/c mice were immunized subcutaneously twice at an interval of three weeks with pcdna3.1 （＋） encoding T cell epitopes of gp63 and Hsp70 individually and in combination. Animals were challenged intracardially with 107 promastigotes ofLeishmania donovani 10 days post immunization and sacrificed 1,2 and 3 months post challenge. The immunized animals revealed a significant reduction （P 〈 0.05） in splenic and hepatic parasite burden as compared to the infected controls. Maximum reduction in parasite load （P 〈 0.05） was observed in animals treated with a combination ofpcdna/gp63 and pcdna/Hsp70. These animals also showed heightened DTH response, increased IgG2a, elevated Thl cytokines （IFN-γ and IL-2） and reduced IgG 1 and IL-10 levels. Thus, mice immunized with the cocktail vaccine exhibited significantly greater protection in comparison to those immunized with individual antigens.

关键词： visceral leishmaniasis dna vaccine T-cell epitopes gp63 Hsp70