High infectivity and unique genomic sequence characteristics of Cryptosporidium parvum in China

Jia, Ruilian; Huang, Wanyi; Huang, Ni; Yu, Zhengjie; Li, Na; Xiao, Lihua; Feng, Yaoyu; Guo, Yaqiong

doi:10.1371/journal.pntd.0010714

Cited by 21 publications

(37 citation statements)

References 34 publications

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“…The IId-HLJ isolate of the IIdA20G1 subtype was initially obtained from a dairy calf on a farm in Heilongjiang (HLJ), China and maintained through passages in GKO mice. This isolate has higher infectivity and some unique genomic sequence characteristics in comparison with the IIaA17G2R1-Waterborne isolate as previously described (25). Oocysts of nonhuman primate-adapted C. hominis isolates of the IbA12G3, ImA20, InA17 subtypes were initially obtained from crab-eating macaques (Macaca fascicularis) in China using sucrose and cesium chloride gradient centrifugations (36,37).…”

Section: Discussionmentioning

confidence: 98%

Cultivation of host-adaptedCryptosporidium parvumandCryptosporidium hominisusing enteroids for cryopreservation of isolates and transcriptomic studies of infection

Deng,

Hou,

Mao

et al. 2023

Preprint

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Cryptosporidium hominisandCryptosporidium parvumare major causes of severe diarrhea in humans. Comparative studies of them are hampered by the lack of effective cultivation and cryopreservation methods, especially forC. hominis. Here, we described adapted murine enteroids for the cultivation of oneC. parvumIId subtype and nonhuman primate-adaptedC. hominisIb, Im, and In subtypes, which allowed the complete development of the pathogens, producing oocysts infectious to mice. Using the system, we developed a novel cryopreservation method forCryptosporidiumisolates. In comparative RNA-seq analyses ofC. hominiscultures, the enteroid system generated significantly more transcriptomic responses of both pathogen and host genes than the conventional HCT-8 cell system. In particular, the infection was shown to upregulate PI3K-Akt, Wnt, Ras,TNF, NF-κB, IL-17, MAPK, and innate immunity signaling pathways and downregulate Wnt and Hippo signaling pathways, host cell metabolism, and parasites in enteroid cultures had significantly higher expression of genes involved in oocyst formation. Therefore, the new culture model provides a valuable tool for comparative studies of the biology of divergentCryptosporidiumspecies.IMPORTANCEThe two dominant species for human cryptosporidiosis,Cryptosporidium hominisandCryptosporidium parvum, differ significantly in host range and virulence. Up to date, biological studies ofCryptosporidiumspp. are almost exclusively done with bovine-adapted IIa subtypes ofC. parvum, which is the species with effective laboratory animal models and in vitro cultivation methods. Here, we describe modified procedures for the generation of murine enteroids for successful cultivation of both nonhuman primate-adaptedC. hominissubtypes and aC. parvumIId subtype, producing oocysts infective to mice. In addition, we have developed a novel cryopreservation method using the system for long-term storage ofCryptosporidiumisolates. RNA-seq analyses ofC. hominiscultures indicate that the enteroid culture system generates host and pathogen transcriptomic responses similar to those in natural infection. This new development alleviates a technical bottleneck in cryptosporidiosis research, and provides an example for other difficult-to-culture pathogens of major public health importance.

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Section: Discussionmentioning

confidence: 98%

Cultivation of host-adaptedCryptosporidium parvumandCryptosporidium hominisusing enteroids for cryopreservation of isolates and transcriptomic studies of infection

Deng,

Hou,

Mao

et al. 2023

Preprint

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“…Differences in host preference and virulence have been reported between GP60 subtype families of C. parvum and C. hominis , respectively (14, 15). Mice experimentally infected with different GP60 subtypes of C. parvum have shown significant differences in infection intensity, duration, and pathogenicity (20). However, few studies have been undertaken to examine the biological processes of GP60-mediated host cell infection.…”

Section: Discussionmentioning

confidence: 99%

Variant Surface Protein GP60 Contributes to Host Infectivity ofCryptosporidium parvum

Li,

Yang,

Hou

et al. 2024

Preprint

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Biological studies of the determinants of Cryptosporidium infectivity and virulence are lacking despite the fact that cryptosporidiosis is a major public health problem. Here we have used advanced genetic tools to investigate the processing, fate and function of the 60-kDa glycoprotein (GP60), an immunodominant variable antigen associated with protection against reinfection. Endogenous gene tagging revealed that GP60 is highly expressed in sporozoites, merozoites and male gametes, suggesting that it may be involved in both invasion and sexual replication. GP60 is translocated to the parasite membrane and cleaved at the furin cleavage site into GP40 and GP15. During invasion, GP40 translocates to the apical end of the zoites and remains detectable at the parasite-host interface. Although GP60 is dispensable, both gene deletion and replacement reduce parasite growth and severity of infection. Depletion of either GP40 or GP15 alone has less effect. These findings advance our understanding of the parasite-host interaction and pathogenesis of cryptosporidiosis.

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“…We have generated genetic crosses of fluorescently tagged C. parvum isolates IIdA20G1-HLJ and IIdA19G1-GD, which differ significantly in infectivity, infection pattern, and virulence in GKO mice. 9 Linkage mapping using BSA of genomes collected over the course of infection with F1 progeny has identified three QTL containing 26 genes that potentially control the virulence difference between the two isolates. Reverse genetic studies of the two best candidates have confirmed the involvement of SKSR1, a secretory small granule protein encoded by a polymorphic subtelomeric gene, in C. parvum virulence.…”

Section: Discussionmentioning

confidence: 99%

“…4,10 However, the genetic determinants of Cryptosporidium virulence and infectivity are not clear, although the results of comparative genomic analysis have provided some clues. 9 In Plasmodium falciparum and Toxoplasma gondii, linkage mapping of genetic crosses of isolates has been effective in the study of virulence factors. 11,12 It has led to the identification of P. falciparum erythrocyte membrane protein 1 (PfEMP1) and several rhoptry proteins as major determinants of virulence in P. falciparum and T. gondii, respectively.…”

Section: Introductionmentioning

confidence: 99%

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SKSR1 identified as key virulence factor inCryptosporidiumby genetic crossing

He,

Sun,

Hou

et al. 2024

Preprint

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Cryptosporidium parvum is a major cause of severe diarrhea. Although isolates of this zoonotic parasite exhibit significant differences in infectivity and virulence, the genetic determinants for these traits are not clear. In this study, we used classical genetics to cross two C. parvum isolates of different virulence and used bulked segregant analysis of whole-genome sequence data from the progeny to identify quantitative trait loci (QTL) associated with Cryptosporidium infectivity and virulence. Of the 26 genes in three QTL, two had loss-of-function mutations in the low-virulence isolates. Deletion of the SKSR1 gene or expression of the frame-shift mutant sequence reduced the pathogenicity of infection in vivo. SKSR1 is a polymorphic secretory protein expressed in small granules and secreted into the parasite-host interface. These results demonstrate that SKSR1 is an important virulence factor in Cryptosporidium, and suggest that this extended family may contribute to pathogenesis.

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High infectivity and unique genomic sequence characteristics of Cryptosporidium parvum in China

Cited by 21 publications

References 34 publications

Cultivation of host-adaptedCryptosporidium parvumandCryptosporidium hominisusing enteroids for cryopreservation of isolates and transcriptomic studies of infection

Cultivation of host-adaptedCryptosporidium parvumandCryptosporidium hominisusing enteroids for cryopreservation of isolates and transcriptomic studies of infection

Variant Surface Protein GP60 Contributes to Host Infectivity ofCryptosporidium parvum

SKSR1 identified as key virulence factor inCryptosporidiumby genetic crossing

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