Department of Dermatology, University of Zurich and University Hospital Zurich, Zurich, Switzerland.
Institute of Medical Microbiology, University of Zurich, Zurich, Switzerland.
Front Immunol. 2022 Jan 31;13:815609. doi: 10.3389/fimmu.2022.815609. eCollection 2022.
Conventional vaccines are very efficient in the prevention of bacterial infections caused by extracellular pathogens due to effective stimulation of pathogen-specific antibodies. In contrast, considering that intracellular surveillance by antibodies is not possible, they are typically less effective in preventing or treating infections caused by intracellular pathogens such as . The objective of the current study was to use so-called photochemical internalization (PCI) to deliver a live bacterial vaccine to the cytosol of antigen-presenting cells (APCs) for the purpose of stimulating major histocompatibility complex (MHC) I-restricted CD8 T-cell responses. For this purpose, BCG (BCG) was combined with the photosensitiser tetraphenyl chlorine disulfonate (TPCS2a) and injected intradermally into mice. TPCS2a was then activated by illumination of the injection site with light of defined energy. Antigen-specific CD4 and CD8 T-cell responses were monitored in blood, spleen, and lymph nodes at different time points thereafter using flow cytometry, ELISA and ELISPOT. Finally, APCs were infected and PCI-treated for analysis of their activation of T cells or after autologous vaccination of mice. Combination of BCG with PCI induced stronger BCG-specific CD4 and CD8 T-cell responses than treatment with BCG only or with BCG and TPCS2a without light. The overall T-cell responses were multifunctional as characterized by the production of IFN-γ, TNF-α, IL-2 and IL-17. Importantly, PCI induced cross-presentation of BCG proteins for stimulation of antigen-specific CD8 T-cells that were particularly producing IFN-γ and TNF-α. PCI further facilitated antigen presentation by causing up-regulation of MHC and co-stimulatory proteins on the surface of APCs as well as their production of TNF-α and IL-1β . Furthermore, PCI-based vaccination also caused local inflammation at the site of vaccination, showing strong infiltration of immune cells, which could contribute to the stimulation of antigen-specific immune responses. This study is the first to demonstrate that a live microbial vaccine can be combined with a photochemical compound and light for cross presentation of antigens to CD8 T cells. Moreover, the results revealed that PCI treatment strongly improved the immunogenicity of BCG.
传统疫苗在预防由细胞外病原体引起的细菌感染方面非常有效,因为它们能有效地刺激针对病原体的抗体。相比之下,由于抗体无法对细胞内进行监测,因此它们在预防或治疗细胞内病原体(如 )引起的感染方面通常效果较差。本研究的目的是使用所谓的光化学内化(PCI)将活细菌疫苗递送至抗原呈递细胞(APC)的细胞质中,以刺激主要组织相容性复合体(MHC)I 受限的 CD8 T 细胞反应。为此,将 BCG(BCG)与光敏剂四苯基氯二砜(TPCS2a)结合,并皮内注射到小鼠体内。然后,用特定能量的光照射注射部位激活 TPCS2a。此后,在不同时间点通过流式细胞术、ELISA 和 ELISPOT 在血液、脾脏和淋巴结中监测抗原特异性 CD4 和 CD8 T 细胞反应。最后,用感染和 PCI 处理的 APC 进行分析,以检测它们对 T 细胞的激活 或 在小鼠自体接种后。BCG 与 PCI 的组合诱导的 BCG 特异性 CD4 和 CD8 T 细胞反应强于单独使用 BCG 或使用 BCG 和无光照的 TPCS2a 处理。总的 T 细胞反应是多功能的,其特征是产生 IFN-γ、TNF-α、IL-2 和 IL-17。重要的是,PCI 诱导了 BCG 蛋白的交叉呈递,以刺激抗原特异性 CD8 T 细胞,这些细胞特别产生 IFN-γ 和 TNF-α。PCI 还通过引起 APC 表面 MHC 和共刺激蛋白的上调以及它们产生 TNF-α 和 IL-1β 来促进抗原呈递 。此外,基于 PCI 的疫苗接种还在接种部位引起局部炎症,表现为免疫细胞的强烈浸润,这可能有助于刺激抗原特异性免疫反应。本研究首次证明,活微生物疫苗可以与光化学化合物和光结合,用于将抗原交叉呈递给 CD8 T 细胞。此外,结果表明 PCI 处理强烈改善了 BCG 的免疫原性。