Protective effect and cellular immune response induced by recombinant Bb-OprH vaccine of Pseudomonas aeruginosa
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摘要:
目的 探讨铜绿假单胞菌(Pa)重组两歧双歧杆菌(rBb)-OprH疫苗免疫及Pa PAO1株攻击后,诱导的小鼠保护力、脾细胞增殖、亚群及凋亡变化情况。 方法 将疫苗口服灌胃免疫Balb/c鼠,在首免后4周用5×107 CFU的PAO1株滴鼻攻击。攻击后2周杀鼠取脾,计数小鼠肺细胞负荷,MTT检测小鼠脾细胞增殖,流式细胞术检测小鼠脾细胞亚群及脾细胞凋亡率。 结果 疫苗免疫及PAO1株攻击后,小鼠肺细菌负荷减少,脾细胞增殖、CD4+ T细胞明显增加,而脾细胞凋亡明显减少。 结论 铜绿假单胞菌重组Bb-OprH疫苗可诱导铜绿假单胞菌感染小鼠产生保护性细胞免疫应答。 -
关键词:
- 铜绿假单胞菌 /
- 重组Bb-OprH疫苗 /
- 脾细胞增殖 /
- 脾细胞亚群 /
- 脾细胞凋亡
Abstract:Objective To investigate protection and changes of splenocyte proliferation, apoptosis and subsets after immunization with recombinant Bifidobacterium (rBb)-OprH vaccine of Pseudomonas aeruginosa (Pa). Methods Balb/c mice were vaccinated intragastrically with the vaccine and challenged by Pa PAO1 strain intranasally at the 4th week after the first immunization. The mice were sacrificed two weeks after the challenge to isolate their lungs and spleens, and the pulmonary bacterial loads were counted. MTT assay was used to detect splenocyte proliferation, while apoptic rate and subsets of splenocytes were measured by flow cytometry. Results After immunization with the vaccine and challenged by Pa PAO1 strain, the bacterial loads in the lungs decreased, while the splenocyte proliferation and the number of CD4+ T cells increased. The apoptic rate decreased. Conclusion The recombinant Bb-OprH vaccine can induce a protective cellular immune response in mice against the Pseudomonas aeruginosa. -
表 1 疫苗免疫及PA01株攻击后小鼠肺组织细菌的菌落数(n=7,Mean±SD)
Group Pulmonary bacterial colonies (lgCFU/g) rBb-OprH group 7.691±0.069* Blank vector control 8.855±0.027 Bb control 8.958±0.037 *P<0.01vs Blank vector control and Bb control. -
[1] Athanasiou CI, Kopsini A. A systematic review on the use of time series data in the study of antimicrobial consumption and Pseudomonas aeruginosa resistance[J]. J Glob Antimicrob Resist, 2018, 15(2): 69-73 [2] Nguyen L, Garcia J, Gruenberg K, et al. Multidrug-Resistant pseudomonas infections: hard to treat, but hope on the horizon[J]. Curr Infect Dis Rep, 2018, 20(8): 1-10 [3] Bassetti M, Vena A, Croxatto A, et al. How to manage Pseudomonas aeruginosa infections[J]. Drugs Context, 2018, 35(7): 212527-35 [4] da Silva AJ, Zangirolami TC, Marques Novo-Mansur MT, et al. Live bacterial vaccine vectors: An overview[J]. Braz J Microbiol, 2014, 45(4): 1117-29 [5] Kucharska I, Liang BY, Ursini N, et al. Molecular interactions of lipopolysaccharide with an outer membrane protein from pseudomonas aeruginosa probed by solution NMR[J]. Biochemistry, 2016, 55(36): 5061-72 [6] Chevalier S, Bouffartigues E, Bodilis JA, et al. Structure, function and regulation of Pseudomonas aeruginosa porins[J]. FEMS Microbiol Rev, 2017, 41(5): 698-722 [7] 徐波, 曹郁生, 陈 燕, 等. 乳酸乳球菌食品级诱导表达系统的构建及异源蛋白的表达[J]. 微生物学报, 2007, 47(4): 604-9 [8] Robinson CM, Kobe BN, Schmitt DM, et al. Genetic engineering of Francisella tularensis LVS for use as a novel live vaccine platform against Pseudomonas aeruginosa infections[J]. Bioengineered, 2015, 6(2): 82-8 [9] Bridge DR, Whitmire JM, Makobongo MO. Heterologous pseudomonas aeruginosa o-antigen delivery using a salmonella enterica serovar typhimurium wecA mutant strain[J]. Int J Med Microbiol, 2016, 306(7): 529-40 [10] Arboleya S, Watkins C, Stanton C, et al. Gut bifidobacteria populations in human health and aging[J]. Front Microbiol, 2016, 7(11): 1204-9 [11] Kitagawa K, Omoto C, Oda T, et al. Oral combination vaccine, comprising bifidobacterium displaying hepatitis C virus nonstructural protein 3 and interferon-alpha, induces strong cellular immunity specific to nonstructural protein 3 in mice[J]. Viral Immunol, 2017, 30(3): 196-203 [12] 刘 潇, 李文桂, 罗广旭. 铜绿假单胞菌重组Bb-pGEX-OprI疫苗的构建及其保护力的研究[J]. 四川大学学报:医学版, 2018, 37(1): 13-7 [13] 朱佑明, 罗永艾, 李文桂. 铜绿假单胞菌重组Bb-OprF疫苗诱导小鼠细胞免疫应答的研究[J]. 免疫学杂志, 2012, 25(3): 217-21 [14] 刘 潇, 李文桂. 两歧双歧杆菌介导的铜绿假单胞菌外膜蛋白Ⅰ(Bb-OprⅠ)疫苗免疫增强小鼠对铜绿假单胞菌的抑制作用[J]. 细胞与分子免疫学杂志, 2017, 28(8): 1040-4 [15] Restagno D, Venet F, Paquet CA, et al. Mice survival and plasmatic cytokine secretion in a "two hit" model of sepsis depend on intratracheal pseudomonas aeruginosa bacterial load[J]. PLoS One, 2016, 11(8): e162109-16 [16] Qadi M, Izquierdo-Rabassa S, Mateu Borras M, et al. Sensing Mg2+ contributes to the resistance of Pseudomonas aeruginosa to complement-mediated opsonophagocytosis[J]. Environ Microbiol, 2017, 19(10, SI): 4278-86 [17] Dunkley ML, Clancy RL, Cripps AW. A role for CD4+ T cells from orally immunized rats in enhanced clearance of Pseudomonas aeruginosa from the lung[J]. Immunology, 1994, 83(3): 362-9 [18] Kamei A, Wu WH, Traficante DC, et al. Collaboration between macrophages and Vaccine-Induced CD4(+) T cells confers protection against lethal pseudomonas aeruginosa pneumonia during neutropenia[J]. J Infect Dis, 2013, 207(1): 39-49 [19] Zhang JL, Jiang R, Wang W, et al. Apoptosis are induced in J774 macrophages upon phagocytosis and killing of Pseudomonas aeruginosa[J]. Cell Immunol, 2013, 286(1/2): 11-5 [20] Hotchkiss RS, Tinsley KW, Swanson PE, et al. Sepsis-induced apoptosis causes progressive profound depletion of B and CD4+ T lymphocytes in humans[J]. J Immunol, 2001, 166(11): 6952-63 [21] Hotchkiss RS, Dunne WM, Swanson PE, et al. Role of apoptosis in Pseudomonas aeruginosa pneumonia[J]. Science, 2001, 294(5548): U1-2 [22] Schreiber T, Swanson PE, Chang KC, et al. Both gram-negative and gram-positive experimental pneumonia induce profound lymphocyte but not respiratory epithelial cell apoptosis[J]. Shock, 2006, 26(3): 271-6 [23] Hotchkiss RS, Tinsley KW, Swanson PE, et al. Prevention of lymphocyte cell death in sepsis improves survival in mice[J]. Proc Natl Acad Sci, 1999, 96(25): 14541-6 [24] Schwulst SJ, Grayson MH, Dipasco PJ, et al. Agonistic monoclonal antibody against CD40 receptor decreases lymphocyte apoptosis and improves survival in sepsis[J]. J Immunol, 2006, 177(1): 557-65 [25] Liang DY, Hou YQ, Lou XL, et al. Decoy receptor 3 improves survival in experimental sepsis by suppressing the inflammatory response and lymphocyte apoptosis[J]. PLoS One, 2015, 10(6): e131680-7