A novel, stain-free, natural auto-fluorescent signal, Sig M, identified from cytometric and transcriptomic analysis of infectivity of Cryptosporidium hominis and Cryptosporidium parvum.

Simple item page
dc.citation.volume13
dc.contributor.authorOgbuigwe P
dc.contributor.authorRoberts JM
dc.contributor.authorKnox MA
dc.contributor.authorHeiser A
dc.contributor.authorPita A
dc.contributor.authorHaack NA
dc.contributor.authorGarcia-Ramirez JC
dc.contributor.authorVelathanthiri N
dc.contributor.authorBiggs PJ
dc.contributor.authorFrench NP
dc.contributor.authorHayman DTS
dc.contributor.editorXu R
dc.coverage.spatialSwitzerland
dc.date.accessioned2024-07-18T03:03:18Z
dc.date.available2024-07-18T03:03:18Z
dc.date.issued2023-05-22
dc.description.abstractCryptosporidiosis is a worldwide diarrheal disease caused by the protozoan Cryptosporidium. The primary symptom is diarrhea, but patients may exhibit different symptoms based on the species of the Cryptosporidium parasite they are infected with. Furthermore, some genotypes within species are more transmissible and apparently virulent than others. The mechanisms underpinning these differences are not understood, and an effective in vitro system for Cryptosporidium culture would help advance our understanding of these differences. Using COLO-680N cells, we employed flow cytometry and microscopy along with the C. parvum-specific antibody Sporo-Glo™ to characterize infected cells 48 h following an infection with C. parvum or C. hominis. The Cryptosporidium parvum-infected cells showed higher levels of signal using Sporo-Glo™ than C. hominis-infected cells, which was likely because Sporo-Glo™ was generated against C. parvum. We found a subset of cells from infected cultures that expressed a novel, dose-dependent auto-fluorescent signal that was detectable across a range of wavelengths. The population of cells that expressed this signal increased proportionately to the multiplicity of infection. The spectral cytometry results confirmed that the signature of this subset of host cells closely matched that of oocysts present in the infectious ecosystem, pointing to a parasitic origin. Present in both C. parvum and C. hominis cultures, we named this Sig M, and due to its distinct profile in cells from both infections, it could be a better marker for assessing Cryptosporidium infection in COLO-680N cells than Sporo-Glo™. We also noted Sig M's impact on Sporo-Glo™ detection as Sporo-Glo™ uses fluoroscein-isothiocynate, which is detected where Sig M also fluoresces. Lastly, we used NanoString nCounter® analysis to investigate the transcriptomic landscape for the two Cryptosporidium species, assessing the gene expression of 144 host and parasite genes. Despite the host gene expression being at high levels, the levels of putative intracellular Cryptosporidium gene expression were low, with no significant difference from controls, which could be, in part, explained by the abundance of uninfected cells present as determined by both Sporo-Glo™ and Sig M analyses. This study shows for the first time that a natural auto-fluorescent signal, Sig M, linked to Cryptosporidium infection can be detected in infected host cells without any fluorescent labeling strategies and that the COLO-680N cell line and spectral cytometry could be useful tools to advance the understanding of Cryptosporidium infectivity.
dc.description.confidentialfalse
dc.edition.edition2023
dc.format.pagination1178576-
dc.identifier.author-urlhttps://www.ncbi.nlm.nih.gov/pubmed/37284498
dc.identifier.citationOgbuigwe P, Roberts JM, Knox MA, Heiser A, Pita A, Haack NA, Garcia-Ramirez JC, Velathanthiri N, Biggs PJ, French NP, Hayman DTS. (2023). A novel, stain-free, natural auto-fluorescent signal, Sig M, identified from cytometric and transcriptomic analysis of infectivity of Cryptosporidium hominis and Cryptosporidium parvum.. Front Cell Infect Microbiol. 13. (pp. 1178576-).
dc.identifier.doi10.3389/fcimb.2023.1178576
dc.identifier.eissn2235-2988
dc.identifier.elements-typejournal-article
dc.identifier.issn2235-2988
dc.identifier.number1178576
dc.identifier.urihttps://mro.massey.ac.nz/handle/10179/70230
dc.languageeng
dc.publisherFrontiers Media S.A.
dc.publisher.urihttps://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2023.1178576/full
dc.relation.isPartOfFront Cell Infect Microbiol
dc.rights(c) 2023 The Author/s
dc.rightsCC BY 4.0
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectcryptosporidiosis
dc.subjectflow cytometry
dc.subjectintracellular infection
dc.subjectnanostring
dc.subjectspectral cytometry
dc.subjectHumans
dc.subjectCryptosporidium
dc.subjectCryptosporidium parvum
dc.subjectCryptosporidiosis
dc.subjectTranscriptome
dc.subjectColoring Agents
dc.subjectEcosystem
dc.subjectDiarrhea
dc.titleA novel, stain-free, natural auto-fluorescent signal, Sig M, identified from cytometric and transcriptomic analysis of infectivity of Cryptosporidium hominis and Cryptosporidium parvum.
dc.typeJournal article
pubs.elements-id461998
pubs.organisational-groupOther
Files
Original bundle
Now showing 1 - 2 of 2
Thumbnail Image
Name:
Published version.pdf
Size:
2.52 MB
Format:
Adobe Portable Document Format
Description:
461998 PDF.pdf
Download
Thumbnail Image
Name:
Evidence.pdf
Size:
1.51 MB
Format:
Adobe Portable Document Format
Description:
Data Sheet 1.PDF
Download
License bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
license.txt
Size:
9.22 KB
Format:
Plain Text
Description:
Download
Collections
Journal Articles