Involvement of INS15 in the development and pathogenicity of the zoonotic pathogen Cryptosporidium parvum.

Autor: He W; State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China.; School of Biology and Agriculture, Shaoguan University, Shaoguan, China., Cui H; State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China., Li N; State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China., Guo Y; State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China., Zeng S; School of Biology and Agriculture, Shaoguan University, Shaoguan, China., Feng Y; State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China., Xiao L; State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China., Xu R; State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, China.
Jazyk: angličtina
Zdroj: PLoS neglected tropical diseases [PLoS Negl Trop Dis] 2024 Oct 03; Vol. 18 (10), pp. e0012569. Date of Electronic Publication: 2024 Oct 03 (Print Publication: 2024).
DOI: 10.1371/journal.pntd.0012569
Abstrakt: Background: Cryptosporidium parvum is a common protozoan pathogen responsible for moderate to severe diarrhea in humans and animals. The C. parvum genome contains 22 genes encoding insulinase-like M16 proteases (INS) with diverse structures and sequences, suggesting that members of the protein family may have distinct biological functions in the life cycle of parasites. Here, we investigated the role of INS15 and INS16, two proteases encoded by neighboring genes with high sequence identity, in the growth and development of C. parvum in vivo and in vitro.
Methodology/principal Findings: INS15 and INS16 genes were tagged and knocked out using CRISPR/Cas9 technology in C. parvum IIdA20G1-HLJ isolate. The expression of INS15 and INS16 was determined by immunofluorescence analysis and immunoelectron microscopy. The effect of depletion of INS15 and INS16 on parasite growth and pathogenicity were assessed on HCT-8 cells and in interferon-γ knockout mice. Endogenous tagging showed that INS15 and INS16 expressed in the oocyst, trophozoite, meront and female gametes. INS15 also expressed in male gamonts, while INS16 was not detected in the male gamonts. Although depletion of the INS15 or INS16 gene affected late development of C. parvum in vitro, only depletion of INS15 significantly reduced parasite burden in infected mice. Mice infected with the INS15-depleted strain had reduced clinical signs, body weight, intestinal villus length to crypt height ratio, and survival time compared to infected with the tagging mutant.
Conclusions/significance: The results of this study indicate that INS15 is mainly involved in the late development of C. parvum. Depletion of this gene attenuates the pathogenicity of this important zoonotic parasite.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 He et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
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