绵羊李斯特菌在小鼠体内的感染增殖规律及诱导的细胞因子分泌
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摘要
目的研究绵羊李斯特菌(Li)静脉接种和滴鼻接种C57BL/6J小鼠后,细菌在小鼠体内感染增殖规律及诱导的细胞免疫应答水平,为Li作为一种新型疫苗载体提供科学依据。方法采用寇氏法测定Li静脉接种和滴鼻接种C57BL/6J小鼠的半数致死剂量(LD_(50));以0.1×LD50剂量静脉或滴鼻接种小鼠,测定不同时间点小鼠肝、脾和肺中的细菌载量、组织病理学改变,以及肝和脾组织匀浆液中IFN-γ、肺组织匀浆液中IFN-γ、TNF-α和IL-6的含量。结果Li静脉接种和滴鼻接种小鼠的半数致死剂量分别为6.3×10~6(cfu)和2.5×10~8(cfu)。Li静脉感染小鼠后能够在肝、脾和肺中增殖,引起组织细胞坏死和炎性浸润并诱导IFN-γ分泌水平上调,三种器官中肝的各指标变化最显著。Li滴鼻感染小鼠后主要在肺部增殖,引起肺部病理损害并诱导肺组织IFN-γ、TNF-α和IL-6分泌水平上调。结论静脉接种Li可以在小鼠体内引起全身性多器官的病理和免疫反应,而滴鼻接种Li可以针对性地在小鼠肺部引起病理和免疫反应;Li可以作为一种新型疫苗载体。
Objective To study the infection and proliferation mechanism and levels of cytokines production in Listeria ivanovii(Li)intravenously or intranasally infected C57BL/6J mice,and to provide scientific basis for using Li as a novel vaccine vector.Methods The median lethal doses of Li for intravenous or intranasal infection of C57 BL/6J mice were determined by Kaeber method.Then mice were infected intravenously or intranasally with Li at the doses of 0.1 × median lethal,bacterial loads and histopathologic changes in the liver,spleen and lung at indicated time points post infection after each inoculation were determined and analyzed,and IFN-γ titers in liver homogenate and spleen homogenate,as well as IFN-γ,TNF-α and IL-6 titers in lung homogenate were determined.Results Median lethal dose of Li for intravenous inoculation and intranasal inoculation were 6.3 × 10~6 cfu and 2.5 × 10~8 cfu respectively.After intravenous inoculation,Li could proliferate in liver,spleen and lung,causing cell destruction and inflammatory infiltration and lead to promoted IFN-γ production,especially in the liver.After intranasal inoculation,Li proliferated mostly in the lung,causing lung tissue damages and promoted production of IFN-γ,TNF-α and IL-6.Conclusion Intravenous inoculation of Li could induce multi-organs pathogenic and immune reactions in mice while intranasal inoculation of Li could only induce notable pathogenic and immune reactions in the lung;Li could be considered as a novel vaccine vector.
Objective To study the infection and proliferation mechanism and levels of cytokines production in Listeria ivanovii(Li)intravenously or intranasally infected C57BL/6J mice,and to provide scientific basis for using Li as a novel vaccine vector.Methods The median lethal doses of Li for intravenous or intranasal infection of C57 BL/6J mice were determined by Kaeber method.Then mice were infected intravenously or intranasally with Li at the doses of 0.1 × median lethal,bacterial loads and histopathologic changes in the liver,spleen and lung at indicated time points post infection after each inoculation were determined and analyzed,and IFN-γ titers in liver homogenate and spleen homogenate,as well as IFN-γ,TNF-α and IL-6 titers in lung homogenate were determined.Results Median lethal dose of Li for intravenous inoculation and intranasal inoculation were 6.3 × 10~6 cfu and 2.5 × 10~8 cfu respectively.After intravenous inoculation,Li could proliferate in liver,spleen and lung,causing cell destruction and inflammatory infiltration and lead to promoted IFN-γ production,especially in the liver.After intranasal inoculation,Li proliferated mostly in the lung,causing lung tissue damages and promoted production of IFN-γ,TNF-α and IL-6.Conclusion Intravenous inoculation of Li could induce multi-organs pathogenic and immune reactions in mice while intranasal inoculation of Li could only induce notable pathogenic and immune reactions in the lung;Li could be considered as a novel vaccine vector.
引文
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[4]Karunasagar I,Krohne G,Goebel W.Listeria ivanovii is capable of cell-to-cell spread involving actin polymerization[J].Infection and Immunity,1993,61(1):162-169.
[5]Lin QQ,Zhou MY,Xu ZK,et al.Construction of two Listeria ivanovii attenuated strains expressing Mycobacterium tuberculosis antigens for TB vaccine purposes[J].Journal of Biotechnology,2015,196:20-26.
[6]Mizuki M,Nakane A,Sekikawa K,et al.Comparison of host resistance to primary and secondary Listeria monocytogenes infections in mice by intranasal and intravenous routes[J].Infection and Immunity,2002,70(9):4805-4811.
[7]Mainou-Fowler T,Macgowan AP,Postlethwaite R.Virulence of listeria spp.:course of infection in resistant and susceptible mice[J].Journal of Medical Microbiology,1988,27(2):131-140.
[8]Hof H,Hefner P.Pathogenicity of listeria monocytogenes in comparison to other listeria species[J].Infection,1988,16(Suppl2):S141-S144.