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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.
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Received: 08 October 2018
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Corresponding Authors:
* wyj@nxu.edu.cn
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[1] 李武, 邓光存, 李敏, 等. 2017. BCG初免-单次重组腺病毒Ad5-CEAB加强免疫策略的免疫效应研究(英文)[J]. 中国人兽共患病学报, 33(6): 501-507.
(Li W, Deng G, Li M, et al.2017. Immune responses induced by BCG prime and single dose of recombinant adenovirus Ad5-CEAB boosted strategy in mice (in English)[J]. Chinese Journal of Zoonoses, 33(6): 501-507.)
[2] Basaraba R J, Izzo A A, Brandt L, et al.2006. Decreased survival of guinea pigs infected with Mycobacterium tuberculosis after multiple BCG vaccinations[J]. Vaccine, 24(3): 280-286.
[3] Bean A G, Roach D R, Briscoe H, et al.1999. Structural deficiencies in granuloma formation in TNF gene-targeted mice underlie the heightened susceptibility to aerosol Mycobacterium tuberculosis infection, which is not compensated for by lymphotoxin[J]. The Journal of Immunology, 162(6): 3504-3511.
[4] Bolger T, O'connell M, Menon A, et al.2006. Complications associated with the Bacille Calmette-Guérin vaccination in Ireland[J]. Archives of Disease in Childhood, 91(7): 594-597.
[5] Buddle B, Wedlock D, Parlane N, et al.2003. Revaccination of neonatal calves with Mycobacterium bovis BCG reduces the level of protection against bovine tuberculosis induced by a single vaccination[J]. Infection and Immunity, 71(11): 6411-6419.
[6] Cooper A, Roberts A, Rhoades Eet al.1995. The role of interleukin-12 in acquired immunity to Mycobacterium tuberculosis infection[J]. Immunology, 84(3): 423-432.
[7] Cooper A M.2009. Cell-mediated immune response in tuberculosis[J]. Annual Review of Immunology, 27: 393-422.
[8] Cooper A M, Khader S A.2008. The role of cytokines in the initiation, expansion, and control of cellular immunity to tuberculosis[J]. Immunological Reviews, 226: 191-204.
[9] Dantas O, De a Ximenes R, De FPM de Albuquerque M, et al.2006. A case-control study of protection against tuberculosis by BCG revaccination in Recife, Brazil[J]. The International Journal of Tuberculosis and Lung Disease, 10(5): 536-541.
[10] De Val B P, Villarreal-Ramos B, Nofrarías M, et al.2012. Goats primed with Mycobacterium bovis BCG and boosted with a recombinant adenovirus expressing Ag85A show enhanced protection against tuberculosis[J]. Clinical and Vaccine Immunology, 19(9): 1339-1347.
[11] Dean G, Whelan A, Clifford D, et al.2014. Comparison of the immunogenicity and protection against bovine tuberculosis following immunization by BCG-priming and boosting with adenovirus or protein based vaccines[J]. Vaccine, 32(11): 1304-1310.
[12] Dou J, Wang Y, Yu F, et al.2012. Protection against Mycobacterium tuberculosis challenge in mice by DNA vaccine Ag85A-ESAT-6-IL-21 priming and BCG boosting[J]. International Journal ofImmunogenetics, 39(2): 183-190.
[13] Fenton M J, Vermeulen M W, Kim S, et al.1997. Induction of gamma interferon production in human alveolar macrophages by Mycobacterium tuberculosis[J]. Infection and Immunity, 65(12): 5149-5156.
[14] Flynn J, Goldstein M M, Triebold K J, et al.1995. IL-12 increases resistance of BALB/c mice to Mycobacterium tuberculosis infection[J]. The Journal of Immunology, 155(5): 2515-2524.
[15] Flynn J L, Chan J.2001. Immunology of tuberculosis[J]. Annual Review of Immunology, 19(1): 93-129.
[16] Flynn J L, Goldstein M M, Chan J, et al.1995. Tumor necrosis factor-α is required in the protective immune response against Mycobacterium tuberculosis in mice[J]. Immunity, 2(6): 561-572.
[17] Hoft D F, Blazevic A, Stanley J, et al.2012. A recombinant adenovirus expressing immunodominant TB antigens can significantly enhance BCG-induced human immunity[J]. Vaccine, 30(12): 2098-2108.
[18] Li W, Deng G, Li M, et al.2014. A recombinant adenovirus expressing CFP10, ESAT6, Ag85A and Ag85B of Mycobacterium tuberculosis elicits strong antigen-specific immune responses in mice[J]. Molecular Immunology, 62(1): 86-95.
[19] Li W, Li M, Deng G, et al.2015. Prime-boost vaccination with Bacillus Calmette Guerin and a recombinant adenovirus co-expressing CFP10, ESAT6, Ag85A and Ag85B of Mycobacterium tuberculosis induces robust antigen-specific immune responses in mice[J]. Molecular Medicine Reports, 12(2): 3073-3080.
[20] Lowrie D B, Tascon R E, Bonato V L, et al.1999. Therapy of tuberculosis in mice by DNA vaccination[J]. Nature, 400(6741): 269-271.
[21] Matucci A, Maggi E, Vultaggio A.2014. Cellular and humoral immune responses during tuberculosis infection: Useful knowledge in the era of biological agents[J]. The Journal of Rheumatology Supplement, 91: 17-23.
[22] Murray P J, Wang L, Onufryk C, et al.1997. T cell-derived IL-10 antagonizes macrophage function in mycobacterial infection[J]. The Journal of Immunology, 158(1): 315-321.
[23] Nuttall J J, Davies M A, Hussey G D, et al.2008. Bacillus Calmette-Guérin (BCG) vaccine-induced complications in children treated with highly active antiretroviral therapy[J]. International Journal of Infectious Diseases, 12(6): e99-e105.
[24] Orlando S, Triulzi I, Ciccacci F, et al.2018. Delayed diagnosis and treatment of tuberculosis in HIV+ patients in Mozambique: A cost-effectiveness analysis of screening protocols based on four symptom screening, smear microscopy, urine LAM test and Xpert MTB/RIF[J]. PLoS One 13(7): e0200523.
[25] Ramshaw I A, Ramsay A J.2000. The prime-boost strategy: Exciting prospects for improved vaccination[J]. Immunology Today, 21(4): 163-165.
[26] Shaban K, Amoudy H A, Mustafa A S.2013. Cellular immune response to recombinant Mycobacterium bovis BCG constructs expressing major antigens of region of difference 1 of Mycobacterium tuberculosis[J]. Clinical and Vaccine Immunology, 20(8): 1230-1237.
[27] Wang J, Wakeham J, Harkness R, et al.1999. Macrophages are a significant source of type 1 cytokines during mycobacterial infection[J]. Journal of Clinical Investigation, 103(7): 1023-1029. |
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