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农业生物技术学报  2019, Vol. 27 Issue (3): 488-494    DOI: 10.3969/j.issn.1674-7968.2019.03.013
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A Single Dose of Recombinant Adenovirus Boosts BCG-primed Specific T Cell Immune Response in Mice (Mus musculus)
LI Wu, DENG Guang-Cun, LIU Xiao-Ming, WANG Yu-Jiong*
Key Laboratory of Ministry of Education for Conservation and Utilization of Special Biological Resources in the Western, Ningxia University, Yinchuan 750021, China
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Abstract  Tuberculosis (TB) is a fatal disease that is caused by Mycobacterium tuberculosis (Mtb). Mycobacterium bovis Bacillus Calmette Guerin (BCG) has been widely used as the unique vaccine for TB prevention for many years, though its protective efficacy against Mtb infection is limited. The development of better vaccines and vaccination strategies to prevent the global spread of Mtb infection is therefore urgently needed. Previous studies had demonstrated that a recombinant adenoviral vector Ad5-CEAB was able to induce robust antigen-specific immune responses in mice (Mus musculus). In this study, antigen-specific lymphocyte proliferation test, interferon-γ enzyme-linked immunospot assay (INF-γ ELISPOT), flow cytometry analysis of splenocytes as well as cytokines enzyme linked immunosorbent assay (ELISA) were used to examine antigen-specific cellular immunological effects of Ad5-CEAB in the mice primed with BCG and boosted with a single dose of the recombinant adenovirus Ad5-CEAB. Female mice of ICR between 6 and 8 weeks of age were randomly divided into 4 groups (8 mice per group), mice in the control groups were injected subcutaneously with phosphate buffer saline (PBS) or with 1×106 CFU of BCG, while mice in the experimental groups were primed subcutaneously with 1×106 CFU of BCG and then intranasally boosted with 1×109 PFU of Ad5-CEAB once or twice. The mice were euthanized 2 weeks after the final immunization for analysis of T cell immune responses.The results displayed significantly elevated splenic T cell proliferation in mice of BCG/Ad5-2 group, BCG/Ad5-1 group and BCG group as compared to the PBS control group (P<0.05). In addition, the results of IFN-γ-ELISPOT also showed a dramatically increased frequency of Mtb antigen-specific IFN-γ-secreting splenic T cells in mice immunized with BCG or BCG/Ad5-CEAB as compared to the PBS-treated mice (P<0.05). The frequencies of CD4+ and CD8+ T cell populations were higher in these immunized mice relative to the PBS-treated group (P<0.05). Noticeably, the above examined indexes of immune responses in mice immunized with one dose of BCG priming and one dose of Ad5-CEAB boosting (BCG/Ad5-1 group) were statistically greater than that in the BCG group and the PBS group (P<0.05). Furthermore, the results of cytokines ELISA showed that significantly elevated antigen-induced cytokines interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α) were found in the BCG/Ad5-1 and BCG/Ad5-2 groups as compared to the BCG and PBS groups (P<0.05). Noticeably, there was no difference in the tested cytokines between the BCG prime-recombinant adenovirus boost groups (P>0.05), an indication that a single dose of recombinant adenovirus boost can effectively stimulate antigen-specific cytokines responses in mice as compared to the BCG alone.These data clearly demonstrate that the single dose of mucosal Ad5-CEAB boost is efficient in stimulating a stronger antigen-specific T cell response in BCG-primed mice as compared to the BCG group as well as PBS control group. Collectively, the results in this study showed that the heterologous prime-boost strategy that subcutaneously primed with BCG and intranasally boosted with a single dose of Ad5-CEAB could elicit robust antigen-specific cellular immune responses in mice. These results provide an additional insight into developing novel Ad-basis TB vaccines and novel mucosal-targeted prime-boost anti-TB vaccination strategies.
Key wordsTuberculosis      Mycobacterium tuberculosis      Prime-boost      Recombinant adenovirus      T cell immunity     
Received: 08 October 2018     
ZTFLH:  S859.79  
Corresponding Authors: * wyj@nxu.edu.cn   
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LI Wu
DENG Guang-Cun
LIU Xiao-Ming
WANG Yu-Jiong
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LI Wu,DENG Guang-Cun,LIU Xiao-Ming, et al. A Single Dose of Recombinant Adenovirus Boosts BCG-primed Specific T Cell Immune Response in Mice (Mus musculus)[J]. 农业生物技术学报, 2019, 27(3): 488-494.
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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2019.03.013     OR     http://journal05.magtech.org.cn/Jwk_ny/EN/Y2019/V27/I3/488
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