Monoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>

ABSTRACT Among the obstacles hindering Cryptosporidium research is the lack of an in vitro culture system that supports complete life development and propagation. This major barrier has led to a shortage of widely available anti-Cryptosporidium antibodies and a lack of markers for staging developmen...

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Autores principales:	Georgia Wilke, Soumya Ravindran, Lisa Funkhouser-Jones, Jennifer Barks, Qiuling Wang, Kelli L. VanDussen, Thaddeus S. Stappenbeck, Theresa B. Kuhlenschmidt, Mark S. Kuhlenschmidt, L. David Sibley
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Cryptosporidium cytoskeleton intracellular parasites membrane proteins monoclonal antibodies sexual development Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c3910f6574d3407b9f6a61d337be9bea
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spelling	oai:doaj.org-article:c3910f6574d3407b9f6a61d337be9bea2021-11-15T15:24:23ZMonoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>10.1128/mSphere.00124-182379-5042https://doaj.org/article/c3910f6574d3407b9f6a61d337be9bea2018-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00124-18https://doaj.org/toc/2379-5042ABSTRACT Among the obstacles hindering Cryptosporidium research is the lack of an in vitro culture system that supports complete life development and propagation. This major barrier has led to a shortage of widely available anti-Cryptosporidium antibodies and a lack of markers for staging developmental progression. Previously developed antibodies against Cryptosporidium were raised against extracellular stages or recombinant proteins, leading to antibodies with limited reactivity across the parasite life cycle. Here we sought to create antibodies that recognize novel epitopes that could be used to define intracellular development. We identified a mouse epithelial cell line that supported C. parvum growth, enabling immunization of mice with infected cells to create a bank of monoclonal antibodies (MAbs) against intracellular parasite stages while avoiding the development of host-specific antibodies. From this bank, we identified 12 antibodies with a range of reactivities across the parasite life cycle. Importantly, we identified specific MAbs that can distinguish different life cycle stages, such as trophozoites, merozoites, type I versus II meronts, and macrogamonts. These MAbs provide valuable tools for the Cryptosporidium research community and will facilitate future investigation into parasite biology. IMPORTANCE Cryptosporidium is a protozoan parasite that causes gastrointestinal disease in humans and animals. Currently, there is a limited array of antibodies available against the parasite, which hinders imaging studies and makes it difficult to visualize the parasite life cycle in different culture systems. In order to alleviate this reagent gap, we created a library of novel antibodies against the intracellular life cycle stages of Cryptosporidium. We identified antibodies that recognize specific life cycle stages in distinctive ways, enabling unambiguous description of the parasite life cycle. These MAbs will aid future investigation into Cryptosporidium biology and help illuminate growth differences between various culture platforms.Georgia WilkeSoumya RavindranLisa Funkhouser-JonesJennifer BarksQiuling WangKelli L. VanDussenThaddeus S. StappenbeckTheresa B. KuhlenschmidtMark S. KuhlenschmidtL. David SibleyAmerican Society for MicrobiologyarticleCryptosporidiumcytoskeletonintracellular parasitesmembrane proteinsmonoclonal antibodiessexual developmentMicrobiologyQR1-502ENmSphere, Vol 3, Iss 3 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cryptosporidium cytoskeleton intracellular parasites membrane proteins monoclonal antibodies sexual development Microbiology QR1-502
spellingShingle	Cryptosporidium cytoskeleton intracellular parasites membrane proteins monoclonal antibodies sexual development Microbiology QR1-502 Georgia Wilke Soumya Ravindran Lisa Funkhouser-Jones Jennifer Barks Qiuling Wang Kelli L. VanDussen Thaddeus S. Stappenbeck Theresa B. Kuhlenschmidt Mark S. Kuhlenschmidt L. David Sibley Monoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>
description	ABSTRACT Among the obstacles hindering Cryptosporidium research is the lack of an in vitro culture system that supports complete life development and propagation. This major barrier has led to a shortage of widely available anti-Cryptosporidium antibodies and a lack of markers for staging developmental progression. Previously developed antibodies against Cryptosporidium were raised against extracellular stages or recombinant proteins, leading to antibodies with limited reactivity across the parasite life cycle. Here we sought to create antibodies that recognize novel epitopes that could be used to define intracellular development. We identified a mouse epithelial cell line that supported C. parvum growth, enabling immunization of mice with infected cells to create a bank of monoclonal antibodies (MAbs) against intracellular parasite stages while avoiding the development of host-specific antibodies. From this bank, we identified 12 antibodies with a range of reactivities across the parasite life cycle. Importantly, we identified specific MAbs that can distinguish different life cycle stages, such as trophozoites, merozoites, type I versus II meronts, and macrogamonts. These MAbs provide valuable tools for the Cryptosporidium research community and will facilitate future investigation into parasite biology. IMPORTANCE Cryptosporidium is a protozoan parasite that causes gastrointestinal disease in humans and animals. Currently, there is a limited array of antibodies available against the parasite, which hinders imaging studies and makes it difficult to visualize the parasite life cycle in different culture systems. In order to alleviate this reagent gap, we created a library of novel antibodies against the intracellular life cycle stages of Cryptosporidium. We identified antibodies that recognize specific life cycle stages in distinctive ways, enabling unambiguous description of the parasite life cycle. These MAbs will aid future investigation into Cryptosporidium biology and help illuminate growth differences between various culture platforms.
format	article
author	Georgia Wilke Soumya Ravindran Lisa Funkhouser-Jones Jennifer Barks Qiuling Wang Kelli L. VanDussen Thaddeus S. Stappenbeck Theresa B. Kuhlenschmidt Mark S. Kuhlenschmidt L. David Sibley
author_facet	Georgia Wilke Soumya Ravindran Lisa Funkhouser-Jones Jennifer Barks Qiuling Wang Kelli L. VanDussen Thaddeus S. Stappenbeck Theresa B. Kuhlenschmidt Mark S. Kuhlenschmidt L. David Sibley
author_sort	Georgia Wilke
title	Monoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>
title_short	Monoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>
title_full	Monoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>
title_fullStr	Monoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>
title_full_unstemmed	Monoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>
title_sort	monoclonal antibodies to intracellular stages of <named-content content-type="genus-species">cryptosporidium parvum</named-content> define life cycle progression <italic toggle="yes">in vitro</italic>
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/c3910f6574d3407b9f6a61d337be9bea
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Monoclonal Antibodies to Intracellular Stages of <named-content content-type="genus-species">Cryptosporidium parvum</named-content> Define Life Cycle Progression <italic toggle="yes">In Vitro</italic>

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