A productive immunocompetent mouse model of cryptosporidiosis with long oocyst shedding duration for immunological studies

He, Xi; Huang, Wanyi; Sun, Lianbei; Hou, Tianyi; Wan, Zhuowei; Li, Na; Guo, Yaqiong; Kváč, Martin; Xiao, Lihua; Feng, Yaoyu

doi:10.1016/j.jinf.2022.02.019

Cited by 11 publications

(12 citation statements)

References 43 publications

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“…This is expected, as the latter consists of one single type of adenocarcinoma cells of the colon compared with diverse populations of intestinal cells of the enteroids. The host cell responses detected in C. hominis -infected ALI cultures, however, are similar to those seen recently in C. parvum and C. tyzzeri -infected ileum (28). The RNA-seq analysis has further shown significant differences in the expression of C. hominis genes between ALI and HCT-8 cultures, with the former having a significantly higher expression of genes encoding proteins associated with oocysts (such as oocyst wall proteins) and sporozoites (such as crystalloid body proteins) (21).…”

Section: Discussionsupporting

confidence: 87%

“…After removing the supernatant, 0.2 ml of Trizol was added into each sample and pipetted several times until the suspension became opaque. The samples were frozen in liquid-nitrogen immediately and stored at -80°C before RNA-seq using the Illumina NovaSeq 6000 at Guangzhou Genedenovo Biotechnology Co. Transcriptomic analysis of the RNA-seq data for host cell responses and C. hominis gene expression was performed as described previously (28). KEGG and GO analyses of DEGs were used to identify biological pathways and terms involved.…”

Section: Methodsmentioning

confidence: 99%

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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: Discussionsupporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

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

Deng,

Hou,

Mao

et al. 2023

Preprint

Self Cite

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show abstract

“…The host cell responses detected in C. hominis -infected ALI cultures, however, are similar to those seen recently in C. parvum and C. tyzzeri -infected ileum. 30 …”

Section: Discussionmentioning

confidence: 99%

“…The samples were frozen in liquid-nitrogen immediately and stored at -80°C before RNA-seq using the Illumina NovaSeq 6000 at Guangzhou Genedenovo Biotechnology Co. Transcriptomic analysis of the RNA-seq data for host cell responses and C. hominis gene expression was performed as described previously. 30 Briefly, the raw sequence reads were assessed for quality control using FastQC v0.11.2. Adapter and low-quality sequences were trimmed using Trimmomatic v0.39.…”

Section: Methodsmentioning

confidence: 99%

Cultivation, cryopreservation, and transcriptomic studies of host-adapted Cryptosporidium parvum and Cryptosporidium hominis using enteroids

Deng,

Hou,

Zhang

et al. 2024

iScience

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“…Previously, differences in infectivity and virulence have been observed between different subtype families of C. parvum and C. tyzzeri [29][30][31] , and between different isolates of the same gp60 subtype [32][33][34] . Results from comparative genomic analysis indicate that sequence differences in genes encoding mucin glycoproteins and other secretory proteins are associated with different phenotypic traits of isolates in mice 29,30 . This is consistent with the data obtained in the present study.…”

Section: Discussionmentioning

confidence: 99%

Unique genomic characteristics underlie the infectivity of divergent Cryptosporidium hominis subtypes to animals

Xiao,

Huang,

et al. 2023

Preprint

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The anthroponotic Cryptosporidium hominis differs from the zoonotic C. parvum in its lack of infectivity to rodents, but several divergent C. hominis subtypes has been recently found in nonhuman primates and equines. Here, we performed whole-genome sequencing of 17 animal C. hominis isolates. In a comparative analysis with 222 human isolates, the animal-adapted isolates showed significant genetic divergence, and genetic recombination shaped their population. In addition, these animal isolates had additional genes found in C. parvum and significant sequence differences in subtelomeric genes. In interferon-γ knockout mice, three monkey isolates showed differences in infectivity and induced higher and longer oocyst shedding than a reference C. parvum isolate. Two of the isolates were fluorescently tagged, allowing the screening of the progeny of a genetic cross. These data provide insight into genetic determinants of host range and virulence in Cryptosporidium, and a convenient animal model for biological studies of C. hominis.

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A productive immunocompetent mouse model of cryptosporidiosis with long oocyst shedding duration for immunological studies

Cited by 11 publications

References 43 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

Cultivation, cryopreservation, and transcriptomic studies of host-adapted Cryptosporidium parvum and Cryptosporidium hominis using enteroids

Unique genomic characteristics underlie the infectivity of divergent Cryptosporidium hominis subtypes to animals

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