Sección de Patología Experimental, Departamento de Patología, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubiran, Mexico City, México.
McMaster Immunology Research Centre & Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada.
Scand J Immunol. 2019 Mar;89(3):e12743. doi: 10.1111/sji.12743. Epub 2019 Jan 24.
High dose of Mycobacterium tuberculosis (Mtb) strain H37Rv administered by intratracheal injection in BALB/c mice induce progressive tuberculosis (TB). In this model, during the first month there is a temporal control of bacillary growth, in coexistence with macrophage activation, granuloma formation and Th-1 response. Then, bacterial proliferation recommences, accompanied by progressive pneumonia and decreasing expression of protective cytokines (IFN-γ and TNF-α). In this model, we studied the IL-12 gene expression kinetics and cellular source. There is a rapid and progressive IL-12 expression peaking at day 14, when granulomas start their formation and numerous macrophages show strong IL-12 immunostaining, while during progressive TB there is a significant decrease of IL-12 expression and occasional macrophages showed IL-12 immunolabeling. In the second part of this study, we determined the immunotherapeutic effect of recombinant adenoviruses that codify IL-12 (AdIL-12). Intratracheal administration of only one dose of AdIL-12 one day before Mtb infection produced significant decrease of bacterial loads, lesser pneumonia and higher expression of TNF-α, IFN-γ and iNOS. When only one dose of AdIL-12 was given in healthy mice cohoused with infected mice with highly virulent and transmissible Mtb, total prevention of infection was conferred. Moreover, when AdIL-12 was administered by intranasal route in animals suffering late active TB after 2 months of infection, a very low pulmonary bacilli burdens was detected. These experimental data confirm that IL-12 is a significant cytokine in the immune protection against Mtb, and gene therapy based in adenoviruses coding this cytokine increased protective immunity and prevent Mtb transmission.
高剂量的结核分枝杆菌(Mtb)菌株 H37Rv 通过气管内注射在 BALB/c 小鼠中诱导进行性肺结核(TB)。在该模型中,在第一个月中存在细菌生长的时间控制,同时存在巨噬细胞激活、肉芽肿形成和 Th-1 反应。然后,细菌增殖重新开始,伴随着进行性肺炎和保护性细胞因子(IFN-γ和 TNF-α)表达的降低。在该模型中,我们研究了 IL-12 基因表达的动力学和细胞来源。IL-12 表达迅速且呈进行性增加,在第 14 天达到峰值,此时肉芽肿开始形成,大量巨噬细胞表现出强烈的 IL-12 免疫染色,而在进行性 TB 中,IL-12 表达显著降低,偶尔有巨噬细胞表现出 IL-12 免疫标记。在本研究的第二部分,我们确定了编码 IL-12 的重组腺病毒(AdIL-12)的免疫治疗效果。在 Mtb 感染前一天仅通过气管内给予一剂 AdIL-12 就可显著降低细菌负荷,减轻肺炎并增加 TNF-α、IFN-γ 和 iNOS 的表达。当仅在与具有高毒力和传染性的 Mtb 感染的小鼠共住的健康小鼠中给予一剂 AdIL-12 时,可完全预防感染。此外,当在感染 2 个月后患有晚期活动性 TB 的动物中通过鼻内途径给予 AdIL-12 时,可检测到肺部细菌载量极低。这些实验数据证实,IL-12 是针对 Mtb 的免疫保护中的重要细胞因子,基于编码该细胞因子的腺病毒的基因治疗可增强保护性免疫并预防 Mtb 传播。