Involvement of a variant secretory protein in virulence of emerging Cryptosporidium parvum subtypes

Several divergent Cryptosporidium parvum subtypes have emerged in people in recent years, but their infectivity, pathogenicity, and genetic characteristics are unclear. In the present study, IFN-γ knockout C57BL/6 (GKO) mice were infected with the novel IIoA15G1 and IIpA11 subtypes of C. parvum and...

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Main Authors: Falei Li, Jiayu Li, Yongping Tang, Wei He, Yingying Fan, Ni Huang, Zhuowei Wan, Martin Kváč, Yaqiong Guo, Na Li, Lihua Xiao, Yaoyu Feng
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:Virulence
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Online Access:https://www.tandfonline.com/doi/10.1080/21505594.2025.2514077
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author Falei Li
Jiayu Li
Yongping Tang
Wei He
Yingying Fan
Ni Huang
Zhuowei Wan
Martin Kváč
Yaqiong Guo
Na Li
Lihua Xiao
Yaoyu Feng
author_facet Falei Li
Jiayu Li
Yongping Tang
Wei He
Yingying Fan
Ni Huang
Zhuowei Wan
Martin Kváč
Yaqiong Guo
Na Li
Lihua Xiao
Yaoyu Feng
author_sort Falei Li
collection DOAJ
description Several divergent Cryptosporidium parvum subtypes have emerged in people in recent years, but their infectivity, pathogenicity, and genetic characteristics are unclear. In the present study, IFN-γ knockout C57BL/6 (GKO) mice were infected with the novel IIoA15G1 and IIpA11 subtypes of C. parvum and the common IIaA17G2R1 subtype. The genomes of these isolates were sequenced and compared with each other. Further gene tagging and deletion were performed on the most polymorphic virulence-associated cgd8_5420 gene encoding a hypothetical protein using the CRISPR/Cas9 technology. IIpA11 and IIoA15G1 were highly infectious in GKO mice, with an ID50 of 2.4 and 3.6 oocysts, respectively. The duration of oocyst shedding for IIpA11 (>58.0 ± 1.4 d) and IIoA15G1 (>57.5 ± 0.9 d) was significantly longer than for IIaA17G2R1 (5.5 ± 0.9 d; p < 0.001). One of the mice infected with IIpA11 died on day 33 post infection. The genomes of IIaA17G2R1, IIoA15G1, and IIpA11 had 203, 46839, and 47,122 single nucleotide polymorphisms, respectively, compared to C. parvum IOWA II. In contrast, only 3,361 nucleotide differences were found between IIoA15G1 and IIpA11, with several genes encoding invasion-associated mucin glycoproteins and cgd8_5420 encoding a secretory protein being highly polymorphic. The latter is mainly expressed in trophozoites, merozoites, and macrogametes. Deletion of this gene reduced the intensity of IIpA11 infection and increased the survival of infected mice. Therefore, the emerging IIoA15G1 and IIpA11 subtypes have divergent genomes compared to common IIa subtypes and are highly infectious and pathogenic in GKO mice. Several secretory proteins, including a variant protein encoded by the subtelomeric cgd8_5420 gene, are associated with differences in virulence between the two subtypes.
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spelling doaj-art-15f82429efed49cba67058c9be5809502025-08-20T03:07:46ZengTaylor & Francis GroupVirulence2150-55942150-56082025-12-0116110.1080/21505594.2025.2514077Involvement of a variant secretory protein in virulence of emerging Cryptosporidium parvum subtypesFalei Li0Jiayu Li1Yongping Tang2Wei He3Yingying Fan4Ni Huang5Zhuowei Wan6Martin Kváč7Yaqiong Guo8Na Li9Lihua Xiao10Yaoyu Feng11State Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaInstitute of Parasitology, Biology Centre of the Academy of Sciences of the Czech Republic, České Budějovice, Czech RepublicState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaState Key Laboratory for Animal Disease Control and Prevention, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong, ChinaSeveral divergent Cryptosporidium parvum subtypes have emerged in people in recent years, but their infectivity, pathogenicity, and genetic characteristics are unclear. In the present study, IFN-γ knockout C57BL/6 (GKO) mice were infected with the novel IIoA15G1 and IIpA11 subtypes of C. parvum and the common IIaA17G2R1 subtype. The genomes of these isolates were sequenced and compared with each other. Further gene tagging and deletion were performed on the most polymorphic virulence-associated cgd8_5420 gene encoding a hypothetical protein using the CRISPR/Cas9 technology. IIpA11 and IIoA15G1 were highly infectious in GKO mice, with an ID50 of 2.4 and 3.6 oocysts, respectively. The duration of oocyst shedding for IIpA11 (>58.0 ± 1.4 d) and IIoA15G1 (>57.5 ± 0.9 d) was significantly longer than for IIaA17G2R1 (5.5 ± 0.9 d; p < 0.001). One of the mice infected with IIpA11 died on day 33 post infection. The genomes of IIaA17G2R1, IIoA15G1, and IIpA11 had 203, 46839, and 47,122 single nucleotide polymorphisms, respectively, compared to C. parvum IOWA II. In contrast, only 3,361 nucleotide differences were found between IIoA15G1 and IIpA11, with several genes encoding invasion-associated mucin glycoproteins and cgd8_5420 encoding a secretory protein being highly polymorphic. The latter is mainly expressed in trophozoites, merozoites, and macrogametes. Deletion of this gene reduced the intensity of IIpA11 infection and increased the survival of infected mice. Therefore, the emerging IIoA15G1 and IIpA11 subtypes have divergent genomes compared to common IIa subtypes and are highly infectious and pathogenic in GKO mice. Several secretory proteins, including a variant protein encoded by the subtelomeric cgd8_5420 gene, are associated with differences in virulence between the two subtypes.https://www.tandfonline.com/doi/10.1080/21505594.2025.2514077Cryptosporidiumvirulencegenomicsanimal modelCRISPR/Cas9
spellingShingle Falei Li
Jiayu Li
Yongping Tang
Wei He
Yingying Fan
Ni Huang
Zhuowei Wan
Martin Kváč
Yaqiong Guo
Na Li
Lihua Xiao
Yaoyu Feng
Involvement of a variant secretory protein in virulence of emerging Cryptosporidium parvum subtypes
Virulence
Cryptosporidium
virulence
genomics
animal model
CRISPR/Cas9
title Involvement of a variant secretory protein in virulence of emerging Cryptosporidium parvum subtypes
title_full Involvement of a variant secretory protein in virulence of emerging Cryptosporidium parvum subtypes
title_fullStr Involvement of a variant secretory protein in virulence of emerging Cryptosporidium parvum subtypes
title_full_unstemmed Involvement of a variant secretory protein in virulence of emerging Cryptosporidium parvum subtypes
title_short Involvement of a variant secretory protein in virulence of emerging Cryptosporidium parvum subtypes
title_sort involvement of a variant secretory protein in virulence of emerging cryptosporidium parvum subtypes
topic Cryptosporidium
virulence
genomics
animal model
CRISPR/Cas9
url https://www.tandfonline.com/doi/10.1080/21505594.2025.2514077
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