弓形虫感染细胞来源的外泌体对慢性鼠弓形虫病的保护作用。

Protective efficacy of Toxoplasma gondii infected cells-derived exosomes against chronic murine toxoplasmosis.

机构信息

Medical Parasitology Department, Faculty of Medicine, Ain Shams University, Cairo, Egypt.

Medical Parasitology Department, Faculty of Medicine, Alexandria University, Alexandria, Egypt.

出版信息

Acta Trop. 2023 Dec;248:107041. doi: 10.1016/j.actatropica.2023.107041. Epub 2023 Oct 17.

DOI:10.1016/j.actatropica.2023.107041

PMID:37858877

Abstract

Exosomes were isolated from T. gondii infected human hepatoblastoma cells using the exosome isolation kit and characterized by electron microscopy and Western blotting. Exosomes adsorbed to alum adjuvant were evaluated as a potential immunizing agent against murine chronic toxoplasmosis compared to excretory secretory antigens (ESA)-alum. Mice were immunized at days 1, 15 and 29. The levels of IgG, IFN-γ, IL-4 and IL-10, CD4 and CD8 T cells were determined using sandwich enzyme-linked immunosorbent assay (sandwich ELISA) at days 14, 28 and 56 of the experiment. Then mice were infected orally with 10 cysts of T. gondii. The protective efficacy of the antigens were evaluated by counting the brain cysts and measuring the aforementioned humoral and cellular parameters 60 days post infection. The results showed that alum increased the protective efficacy of the exosomes. Immunization with exosome-alum induced both humoral and mixed Th1/Th2 cellular immune responses. Exosome-alum gave higher levels of the humoral and cellular parameters, compared to ESA-alum. After challenge infection, exosome-alum significantly reduced the brain cyst burden by 75 % while ESA-alum gave 42 % reduction and evoked higher humoral and cellular immune responses. Therefore, the possibility of using T. gondii infected cells-derived exosome-alum as a vaccine is a new perspective in toxoplasmosis.

摘要

从 T. gondii 感染的人肝癌细胞中使用外泌体分离试剂盒分离外泌体，并通过电子显微镜和 Western blot 进行表征。将吸附在明矾佐剂上的外泌体与排泄分泌抗原（ESA）-明矾进行比较，作为针对慢性弓形虫病的潜在免疫原进行评估。在第 1、15 和 29 天给小鼠免疫。在实验的第 14、28 和 56 天，使用夹心酶联免疫吸附测定（夹心 ELISA）测定 IgG、IFN-γ、IL-4 和 IL-10、CD4 和 CD8 T 细胞的水平。然后用 10 个 T. gondii 囊虫口服感染小鼠。通过计算脑囊虫和测量感染后 60 天上述体液和细胞参数来评估抗原的保护效力。结果表明，明矾增加了外泌体的保护效力。用外泌体-明矾免疫诱导体液和混合 Th1/Th2 细胞免疫应答。与 ESA-明矾相比，外泌体-明矾诱导产生更高水平的体液和细胞参数。在挑战感染后，外泌体-明矾可使脑囊虫负担减少 75%，而 ESA-明矾则减少 42%，并引起更高的体液和细胞免疫应答。因此，使用 T. gondii 感染细胞衍生的外泌体-明矾作为疫苗的可能性是弓形虫病的一个新视角。