Characterization of Membrane-Associated Progesterone Receptor Component-2 (MAPRC2) from Trichinella spiralis and Its Interaction with Progesterone and Mifepristone
Trichinellosis is a foodborne zoonotic disease caused by Trichinella spp., including Trichinella spiralis. In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized using protein sequencing analysis. Furthermore, the express...
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| Veröffentlicht in: | Vaccines (Basel) 2021-08, Vol.9 (8), p.934 |
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| creator | Aleem, Muhammad Tahir Shi, Jiawen Yu, Zhengqing Wen, Zhaohai Zhang, Yang Liang, Meng Lakho, Shakeel Ahmed Haseeb, Muhammad Ali, Haider Hassan, Muhammad Waqas Song, Xiaokai Li, Xiangrui Xu, Lixin Yan, Ruofeng |
| description | Trichinellosis is a foodborne zoonotic disease caused by Trichinella spp., including Trichinella spiralis. In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized using protein sequencing analysis. Furthermore, the expression, purification, immunoblot assay, binding ability with progesterone antibody, and immunofluorescence assay were performed. A direct effect of progesterone (P4) and mifepristone (RU486) on the Ts-MAPRC2 gene was determined using in vitro cell culture that showed different expression levels at all developmental stages (muscle larvae (ML), female adult worm (F-AL), male adult worm (M-AL), and newborn larvae (NBL)). Subsequently, the in vitro phenotypic effects of P4, RU486, and rTs-MAPRC2-Ab on F-AL and ML stages were measured. Later, the in vivo phenotypic effect and relative mRNA expression of mifepristone on the F-AL stage were studied. Our results revealed that the Ts-MAPRC2 gene is critical to maintaining pregnancy in the female adult worm (F-AL) of T. spiralis. The 300 ng/mL of P4 and 100 ng/mL of RU486 showed downregulation of the Ts-MAPRC2 gene in F-AL (p ≤ 0.05). This plays an important role in abortion and possibly decreases the worm burden of T. spiralis in the host. Only 30 ng/mL P4 showed significant upregulation in F-AL (p ≤ 0.05). The current study provides new insights regarding the antihormone (P4 and RU486) drug design and vaccine therapy of recombinant (rTs-MAPRC2) protein as well as their combined effects to control T. spiralis infection. |
| doi_str_mv | 10.3390/vaccines9080934 |
| format | Article |
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In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized using protein sequencing analysis. Furthermore, the expression, purification, immunoblot assay, binding ability with progesterone antibody, and immunofluorescence assay were performed. A direct effect of progesterone (P4) and mifepristone (RU486) on the Ts-MAPRC2 gene was determined using in vitro cell culture that showed different expression levels at all developmental stages (muscle larvae (ML), female adult worm (F-AL), male adult worm (M-AL), and newborn larvae (NBL)). Subsequently, the in vitro phenotypic effects of P4, RU486, and rTs-MAPRC2-Ab on F-AL and ML stages were measured. Later, the in vivo phenotypic effect and relative mRNA expression of mifepristone on the F-AL stage were studied. Our results revealed that the Ts-MAPRC2 gene is critical to maintaining pregnancy in the female adult worm (F-AL) of T. spiralis. The 300 ng/mL of P4 and 100 ng/mL of RU486 showed downregulation of the Ts-MAPRC2 gene in F-AL (p ≤ 0.05). This plays an important role in abortion and possibly decreases the worm burden of T. spiralis in the host. Only 30 ng/mL P4 showed significant upregulation in F-AL (p ≤ 0.05). The current study provides new insights regarding the antihormone (P4 and RU486) drug design and vaccine therapy of recombinant (rTs-MAPRC2) protein as well as their combined effects to control T. spiralis infection.</description><identifier>ISSN: 2076-393X</identifier><identifier>EISSN: 2076-393X</identifier><identifier>DOI: 10.3390/vaccines9080934</identifier><identifier>PMID: 34452060</identifier><language>eng</language><publisher>Basel: MDPI AG</publisher><subject>Animals ; Antibodies ; By products ; Cell culture ; Cloning ; Combined vaccines ; Developmental stages ; Drug development ; Drug dosages ; Drug resistance ; Enzymes ; Females ; Gene expression ; hormones ; Immunofluorescence ; Infections ; Larvae ; MAPRC2 ; Membranes ; Mifepristone ; Muscles ; Parasites ; Parasitic diseases ; Progesterone ; progesterone receptor ; Protein purification ; Proteins ; Receptors ; Sequence analysis ; Trichinella spiralis ; Trichinellosis ; Vaccines ; Zoonoses</subject><ispartof>Vaccines (Basel), 2021-08, Vol.9 (8), p.934</ispartof><rights>2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2021 by the authors. 2021</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c464t-1320bba67b2234fd4599174a9a84f393d2112d439e0a5167b57fb4b6c8a627023</citedby><cites>FETCH-LOGICAL-c464t-1320bba67b2234fd4599174a9a84f393d2112d439e0a5167b57fb4b6c8a627023</cites><orcidid>0000-0003-2305-2092 ; 0000-0002-7143-6827 ; 0000-0001-8899-2777 ; 0000-0002-2587-0171</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8402905/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8402905/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,861,882,2096,27905,27906,53772,53774</link.rule.ids></links><search><creatorcontrib>Aleem, Muhammad Tahir</creatorcontrib><creatorcontrib>Shi, Jiawen</creatorcontrib><creatorcontrib>Yu, Zhengqing</creatorcontrib><creatorcontrib>Wen, Zhaohai</creatorcontrib><creatorcontrib>Zhang, Yang</creatorcontrib><creatorcontrib>Liang, Meng</creatorcontrib><creatorcontrib>Lakho, Shakeel Ahmed</creatorcontrib><creatorcontrib>Haseeb, Muhammad</creatorcontrib><creatorcontrib>Ali, Haider</creatorcontrib><creatorcontrib>Hassan, Muhammad Waqas</creatorcontrib><creatorcontrib>Song, Xiaokai</creatorcontrib><creatorcontrib>Li, Xiangrui</creatorcontrib><creatorcontrib>Xu, Lixin</creatorcontrib><creatorcontrib>Yan, Ruofeng</creatorcontrib><title>Characterization of Membrane-Associated Progesterone Receptor Component-2 (MAPRC2) from Trichinella spiralis and Its Interaction with Progesterone and Mifepristone</title><title>Vaccines (Basel)</title><description>Trichinellosis is a foodborne zoonotic disease caused by Trichinella spp., including Trichinella spiralis. In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized using protein sequencing analysis. Furthermore, the expression, purification, immunoblot assay, binding ability with progesterone antibody, and immunofluorescence assay were performed. A direct effect of progesterone (P4) and mifepristone (RU486) on the Ts-MAPRC2 gene was determined using in vitro cell culture that showed different expression levels at all developmental stages (muscle larvae (ML), female adult worm (F-AL), male adult worm (M-AL), and newborn larvae (NBL)). Subsequently, the in vitro phenotypic effects of P4, RU486, and rTs-MAPRC2-Ab on F-AL and ML stages were measured. Later, the in vivo phenotypic effect and relative mRNA expression of mifepristone on the F-AL stage were studied. Our results revealed that the Ts-MAPRC2 gene is critical to maintaining pregnancy in the female adult worm (F-AL) of T. spiralis. The 300 ng/mL of P4 and 100 ng/mL of RU486 showed downregulation of the Ts-MAPRC2 gene in F-AL (p ≤ 0.05). This plays an important role in abortion and possibly decreases the worm burden of T. spiralis in the host. Only 30 ng/mL P4 showed significant upregulation in F-AL (p ≤ 0.05). 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receptor</subject><subject>Protein purification</subject><subject>Proteins</subject><subject>Receptors</subject><subject>Sequence analysis</subject><subject>Trichinella 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of Membrane-Associated Progesterone Receptor Component-2 (MAPRC2) from Trichinella spiralis and Its Interaction with Progesterone and Mifepristone</atitle><jtitle>Vaccines (Basel)</jtitle><date>2021-08-23</date><risdate>2021</risdate><volume>9</volume><issue>8</issue><spage>934</spage><pages>934-</pages><issn>2076-393X</issn><eissn>2076-393X</eissn><abstract>Trichinellosis is a foodborne zoonotic disease caused by Trichinella spp., including Trichinella spiralis. In the present study, T. spiralis membrane-associated progesterone receptor component-2 (Ts-MAPRC2) gene was cloned and characterized using protein sequencing analysis. Furthermore, the expression, purification, immunoblot assay, binding ability with progesterone antibody, and immunofluorescence assay were performed. A direct effect of progesterone (P4) and mifepristone (RU486) on the Ts-MAPRC2 gene was determined using in vitro cell culture that showed different expression levels at all developmental stages (muscle larvae (ML), female adult worm (F-AL), male adult worm (M-AL), and newborn larvae (NBL)). Subsequently, the in vitro phenotypic effects of P4, RU486, and rTs-MAPRC2-Ab on F-AL and ML stages were measured. Later, the in vivo phenotypic effect and relative mRNA expression of mifepristone on the F-AL stage were studied. Our results revealed that the Ts-MAPRC2 gene is critical to maintaining pregnancy in the female adult worm (F-AL) of T. spiralis. The 300 ng/mL of P4 and 100 ng/mL of RU486 showed downregulation of the Ts-MAPRC2 gene in F-AL (p ≤ 0.05). This plays an important role in abortion and possibly decreases the worm burden of T. spiralis in the host. Only 30 ng/mL P4 showed significant upregulation in F-AL (p ≤ 0.05). The current study provides new insights regarding the antihormone (P4 and RU486) drug design and vaccine therapy of recombinant (rTs-MAPRC2) protein as well as their combined effects to control T. spiralis infection.</abstract><cop>Basel</cop><pub>MDPI AG</pub><pmid>34452060</pmid><doi>10.3390/vaccines9080934</doi><orcidid>https://orcid.org/0000-0003-2305-2092</orcidid><orcidid>https://orcid.org/0000-0002-7143-6827</orcidid><orcidid>https://orcid.org/0000-0001-8899-2777</orcidid><orcidid>https://orcid.org/0000-0002-2587-0171</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | Vaccines (Basel), 2021-08, Vol.9 (8), p.934 |
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| source | DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central Open Access; MDPI - Multidisciplinary Digital Publishing Institute; PubMed Central |
| subjects | Animals Antibodies By products Cell culture Cloning Combined vaccines Developmental stages Drug development Drug dosages Drug resistance Enzymes Females Gene expression hormones Immunofluorescence Infections Larvae MAPRC2 Membranes Mifepristone Muscles Parasites Parasitic diseases Progesterone progesterone receptor Protein purification Proteins Receptors Sequence analysis Trichinella spiralis Trichinellosis Vaccines Zoonoses |
| title | Characterization of Membrane-Associated Progesterone Receptor Component-2 (MAPRC2) from Trichinella spiralis and Its Interaction with Progesterone and Mifepristone |
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