Xu Mengfei, Zhang Xichen, Yu Qile, Zhao Qi, Li Jianhua, Gong Pengtao, Wang Xiaocen, Li Xin, Zhang Xu, Zhang Nan
State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun 130062, China.
State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun 130062, China.
Vet Parasitol. 2025 Aug;338:110539. doi: 10.1016/j.vetpar.2025.110539. Epub 2025 Jul 9.
Cryptosporidium parvum (C. parvum) is a common zoonotic protozoan pathogen that can cause severe diarrhea in both humans and animals. However, the factors involved in its pathogenicity remain incompletely understood. The C. parvum genome contains nine genes that encode peptidyl-prolyl cis/trans isomerases (PPIases). Previous bioinformatics analyses have indicated that proteins within this family may be associated with the pathogenicity of C. parvum. Here, we explored the role of cyclophilin 23 (CpCyP23), a member of the PPIase family, in the development and pathogenicity of C. parvum. In this study, the CpCyP23 gene was tagged and deleted using CRISPR/Cas9 technology in C. parvum. Immunofluorescence analysis demonstrated that CpCyP23 is expressed in all key developmental stages of C. parvum. The impact of CpCyP23 deficiency on parasite development and pathogenicity were assessed in HCT-8 cells and interferon-γ knockout mice, and the results revealed that the lack of CpCyP23 delayed the development of C. parvum in vitro. Moreover, compared with mice inoculated with the tagged strain, those infected with the knockout strain exhibited a reduction in parasite burden and small intestinal damage. These findings demonstrate that CpCyP23 plays a role in the development of C. parvum, and the deletion of the CpCyP23 gene reduces the pathogenicity of the C. parvum. Overall, these results advance our understanding of the pathogenic mechanisms of C. parvum and suggest CpCyP23 is a promising target for intervention in cryptosporidiosis.
微小隐孢子虫（C. parvum）是一种常见的人畜共患原生动物病原体，可导致人类和动物严重腹泻。然而，其致病相关因素仍未完全明确。微小隐孢子虫基因组包含九个编码肽基脯氨酰顺/反异构酶（PPIases）的基因。先前的生物信息学分析表明，该家族中的蛋白质可能与微小隐孢子虫的致病性有关。在此，我们探究了PPIase家族成员亲环素23（CpCyP23）在微小隐孢子虫发育和致病性中的作用。在本研究中，利用CRISPR/Cas9技术在微小隐孢子虫中对CpCyP23基因进行标记和缺失。免疫荧光分析表明，CpCyP23在微小隐孢子虫的所有关键发育阶段均有表达。在HCT-8细胞和干扰素-γ基因敲除小鼠中评估了CpCyP23缺失对寄生虫发育和致病性的影响，结果显示，缺乏CpCyP23会延迟微小隐孢子虫在体外的发育。此外，与接种标记菌株的小鼠相比，感染基因敲除菌株的小鼠体内寄生虫负荷降低，小肠损伤减轻。这些发现表明，CpCyP23在微小隐孢子虫的发育中发挥作用，CpCyP23基因的缺失降低了微小隐孢子虫的致病性。总体而言，这些结果增进了我们对微小隐孢子虫致病机制的理解，并表明CpCyP23是隐孢子虫病干预的一个有前景的靶点。
