摘要:
The present study aimed to determine the intracellular localization of Cpn0425 in Chlamydophila pneumoniae-infected cells. The recombinant plasmid pGEX-6p/Cpn0425 was transformed into E.coli Bl21 cells to express the fusion protein. Following purification with glutathione S-transferase (GST) resin chromatography, the Cpn0425 fusion protein was used to induce immunity in mice to develop monoclonal and polyclonal antibodies, which were subsequently used to localize the endogenous Cpn0425 protein by indirect immunofluorescence assay (IFA). ELISA was used to determine the immunogenicity of the Cpn0425 plasmid protein by recognizing the pool sera of patients infected with Chlamydia trachomatis and the pool sera of mice immunized with the Cpn0425 fusion protein. The Cpn0425 gene was expressed as the GST-Cpn0425 fusion protein in E. coli and its antibody was prepared by immunizing mice with the fusion protein. An anti-GST-Cpn0425 antibody was used to localize the protein in cells infected with Chlamydophila pneumoniae AR-39 using an IFA. The anti-GST-CT058 antibody detected an inclusion signal in the IFA. Cpn0425 protein strongly reacted with antiserum. Although Cpn0425 protein is not a secreted protein, it has good immunogenicity. Therefore, this protein may be useful for developing vaccines against Chlamydophila pneumoniae infection.
作者机构:
[Lu, Chunxue; Li, Zhihong; Lei, Lei; Zeng, Hao; Zhong, Guangming] Univ Texas Hlth Sci Ctr San Antonio, Dept Microbiol & Immunol, San Antonio, TX 78229 USA.;[Yeh, I-Tien] Univ Texas Hlth Sci Ctr San Antonio, Dept Pathol, San Antonio, TX 78229 USA.;[Lu, Chunxue; Wu, Yimou] Univ S China, Dept Microbiol & Immunol, Hengyang 421001, Hunan, Peoples R China.;[Zhong, Guangming] Univ Texas Hlth Sci Ctr San Antonio, Dept Microbiol & Immunol, 7703 Floyd Curl Dr, San Antonio, TX 78229 USA.
通讯机构:
[Zhong, Guangming] U;Univ Texas Hlth Sci Ctr San Antonio, Dept Microbiol & Immunol, 7703 Floyd Curl Dr, San Antonio, TX 78229 USA.
关键词:
Chlamydia muridarum;Live infection;Pathology;IFN gamma and IL-17;Secretion proteins
摘要:
To search for optimal immunization conditions for inducing protective immunity against upper genital tract pathologies caused by chlamydial intravaginal infection, we compared protection efficacy in mice immunized intranasally or intramuscularly with live or inactivated Chlamydia muridarum organisms. Mice immunized intranasally with live organisms developed strong protection against both vaginal shedding of infectious organisms and upper genital tract pathologies. The protection correlated with a robust antigen-specific T cell response with high IFN gamma but low IL-17. Although a significant level of IL-5 was also detected, these mice maintained an overall Th1-dorminant immunity following immunization and challenge infection. On the contrary, mice immunized intranasally with inactivated organisms or intramuscularly with live or inactivated organisms produced high levels of IL-17 and still developed significant upper genital tract pathologies. High titers of antibodies against chlamydial secretion antigens were detected only in mice immunized intranasally with live organisms but not mice in other groups, suggesting that the intranasally inoculated live organisms were able to undergo replication and immune responses to the chlamydial secretion proteins may contribute to protective immunity. These observations have provided important information on how to develop subunit vaccines for inducing protective immunity against urogenital infection with Chlamydia trachomatis organisms. (C) 2011 Elsevier Ltd. All rights reserved.
期刊:
CANADIAN JOURNAL OF MICROBIOLOGY,2012年58(5):644-652 ISSN:0008-4166
通讯作者:
Wang, Shiping
作者机构:
[Wang, Shiping; Zhu, Cuiming] Cent S Univ, Xiangya Sch Med, Changsha 410078, Hunan, Peoples R China.;[Wu, Yimou; Xiao, Jinhong; Yu, Minjun; You, Xiaoxing; Zeng, Yanhua; Chen, Sufang] Univ S China, Dept Microbiol & Immunol, Hengyang 421001, Peoples R China.
通讯机构:
[Wang, Shiping] C;Cent S Univ, Xiangya Sch Med, Changsha 410078, Hunan, Peoples R China.
关键词:
Mycoplasma pneumoniae;P1 adhesin protein carboxy terminal region;DNA vaccine;immune response;Mycoplasma pneumoniae;région carboxy terminale de l’adhésine P1;vaccin à ADN;réponse immune
摘要:
Mycoplasma pneumoniae is an important causative agent of atypical pneumonia. This study was to determine the ability of a DNA expression vector, which encodes the carboxy terminal region of the M. pneumoniae P1 protein (P1C), to induce humoral and cellular immune responses and to protect against M. pneumoniae infection in BALB/c mice. Mice were immunized with pcDNA3.1/P1C by either intramuscular injection (i.m.) or intranasal inoculation (i.n.). Our results showed that p1c DNA immunization generates detectable antibodies specific to M. pneumoniae, and elicits high levels of IgG1, IgG2a, and IgG2b isotypes (P < 0.01). The levels of IFN-γ and IL-4 in spleen cells of the immunized mice were significantly elevated by immunization via both the i.m. and i.n. methods. Moreover, p1c DNA-immunized mice exhibited detectable protection against M. pneumoniae infection. The lung tissue inflammation was relieved and the histopathologic score (HPS) of pcDNA3.1/P1C-immunized mice was significantly decreased than those in phosphate-buffed saline (PBS) or vaccine-vector-immunized mice (P < 0.01), whereas there were no significant differences in HPS between i.m. and i.n. vaccination (P > 0.05). Our results suggest that pcDNA3.1/P1C could be useful for developing a vaccine against M. pneumoniae infection.
作者机构:
[陆春雪; 吴移谋; 李忠玉; 胡四海] 南华大学微生物与免疫学教研室;[彭波] 南华大学肿瘤研究所;[钟光明] Department of Microbiology and Immunology,University of Texas Health Science Center at San Antonio
作者机构:
[Zhao Feijun; Zeng Tiebing; Yu Jian; Chen Xi; Wu Yimou] Pathogenic Biology Institute,University of South China,Hengyang City,Hunan Province 421001,China;[Liu Shuangquan] Department of Anatomy & Neurobiology,Xiangya School of Medicine,Central South University,Changsha,410013,China;[Liu Shuangquan] First Affiliated Hospital,University of South China,Hengyang City,Hunan Province 411000,China;[Zhang Xiaohong] Department of Histology and Embryology,School of Medicine,University of South China,Hengyang City,Hunan Province 421001,China;[Gu Weiming] Skin Diseases and STD Hospital,Shanghai 200050,China
作者机构:
[Zhou Zhou; Zhang Jun Hua; Liu Guang Chao; Chen Li Li; Zhou An Wen; Wu Yi Mou; Liu Liang Zhuan] Univ S China, Dept Microbiol & Immunol, Hengyang 421001, Hunan, Peoples R China.
通讯机构:
[Wu Yi Mou] U;Univ S China, Dept Microbiol & Immunol, Hengyang 421001, Hunan, Peoples R China.
摘要:
In the present study, immunomodulatory responses of a DNA vaccine constructed by fusing Treponema pallidum (Tp) glycerophosphodiester phosphodiesterase (Gpd) to interleukin-2 (IL-2) and using chitosan (CS) nanoparticles as vectors were investigated. New Zealand white rabbits were immunized by intramuscular inoculation of control DNAs, Tp Gpd DNA vaccine, or Gpd-IL-2 fusion DNA vaccine, which were vectored by CS nanoparticles. Levels of the anti-Gpd antibodies and levels of IL-2 and interferon-gamma in rabbits were increased upon inoculation of Gpd-IL-2 fusion DNA vaccine, when compared with the inoculation with Gpd DNA vaccine, with CS vectoring increasing the effects. The Gpd-IL-2 fusion DNA vaccine efficiently enhanced the antigen-specific lymphocyte proliferative response. When the rabbits were challenged intradermally with 10(5) Tp (Nichols) spirochetes, the Gpd-IL-2 fusion DNA vaccine conferred better protection than the Gpd DNA vaccine (P < 0.05), as characterized by lower detectable amounts of dark field positive lesions (17.5%), lower ulcerative lesion scores (15%), and faster recovery. Individuals treated with the Tp Gpd-IL-2 fusion DNA vaccine vectored by CS nanoparticles had the lowest amounts of dark field positive lesions (10%) and ulcerations (5%) observed and the fastest recovery (42 days). These results indicate that the Gpd-IL-2 fusion DNA vaccine vectored by CS nanoparticles can efficiently induce Th1-dominant immune responses, improve protective efficacy against Tp spirochete infection, and effectively attenuate development of syphilitic lesions.
作者机构:
[Zhou Hui; Li Zhongyu; Wu Yimou; Ma Kangkang; Zhou Zhou; Lu Chunxue] Pathogenic Biology Institute,University of South China,Hengyang 421001,China
通讯机构:
[Wu, Yimou] U;Univ S China, Pathogen Biol Inst, Dept Microbiol & Immunol, Hengyang 421001, Peoples R China.
关键词:
Mycoplasma pneumoniae;P1 adhesin protein carboxy terminal region;the B subunit of Escherichia coliheat-labile enterotoxin;DNA vaccine;Mycoplasma pneumoniae;région carboxyterminale de l’adhésine P1;sous-unité B de l’entérotoxine thermolabile d’Escherichia coli;vaccin à ADN
摘要:
In the present study, we investigated the immunomodulatory responses of a DNA vaccine constructed by fusing Mycoplasma pneumoniae P1 protein carboxy terminal region (P1C) with the Escherichia coli heat-labile toxin B subunit (LTB). BALB/c mice were immunized by intranasal inoculation with control DNAs, the P1C DNA vaccine or the LTB-P1C fusion DNA vaccine. Levels of the anti-M. pneumoniae antibodies and levels of interferon-gamma and IL-4 in mice were increased significantly upon inoculation of the LTB-P1C fusion DNA vaccine when compared with the inoculation with P1C DNA vaccine. The LTB-P1C fusion DNA vaccine efficiently enhanced the M. pneumoniae-specific IgA and IgG levels. The IgG2a/IgG1 ratio was significantly higher in bronchoalveolar lavages fluid and sera from mice fusion with LTB and P1C than mice receiving P1C alone. When the mice were challenged intranasally with 10(7) CFU M. pneumoniae strain (M129), the LTB-P1C fusion DNA vaccine conferred significantly better protection than P1C DNA vaccine (P < 0.05), as suggested by the results, such as less inflammation, lower histopathological score values, lower detectable number of M. pneumoniae strain, and lower mortality of challenging from 5 x 10(8) CFU M. pneumoniae. These results indicated that the LTB-P1C fusion DNA vaccine efficiently improved protective efficacy against M. pneumoniae infection and effectively attenuated development of M. pneumoniae in mice.