Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay

Proliferative enteropathy (PE) is an infectious enteric disease caused by Lawsonia intracellularis (L. intracellularis) and is endemic in pig herds worldwide. However, a L. intracellularis-specific monoclonal antibody plays an important role in the evaluation of L. intracellularis infection in vitro...

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Autores principales:	Ning Xiao, Jiannan Li, Minxue Li, Yuting Hu, Huixing Lin, Hongjie Fan
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	proliferative enteropathy Lawsonia intracellularis Omp2 protein monoclonal antibody indirect immunofluorescence assay immunocytochemistry Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/7f326a1807b544d4b5ec8d19701e92d4
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spelling	oai:doaj.org-article:7f326a1807b544d4b5ec8d19701e92d42021-12-01T13:40:59ZPreparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay2297-176910.3389/fvets.2021.753610https://doaj.org/article/7f326a1807b544d4b5ec8d19701e92d42021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fvets.2021.753610/fullhttps://doaj.org/toc/2297-1769Proliferative enteropathy (PE) is an infectious enteric disease caused by Lawsonia intracellularis (L. intracellularis) and is endemic in pig herds worldwide. However, a L. intracellularis-specific monoclonal antibody plays an important role in the evaluation of L. intracellularis infection in vitro. Therefore, the objective of this study was to produce and identify the characteristics of a new monoclonal antibody against the outer membrane protein (Omp2) of L. intracellularis and apply it in an indirect immunofluorescence assay (IFA) and immunocytochemistry (IHC). The results indicated that three highly specific monoclonal antibodies against the Omp2 protein (4D9, 3G2, and 7G5) of L. intracellularis were obtained by using purified Omp2 as an immunogen, the titers of ascitic fluids of 4D9, 3G2, and 7G5 cells were 1:2,048,000, 1:512,000, and 1:256,000, respectively. IFA analysis showed that the 4D9, 3G2, and 7G5 have no cross-reactivity with other enteric bacteria commonly found in the ilea of pigs or closely related to L. intracellularis, such as Desulfovibrio, Bilophila wadsworthia (B. wadsworthia), Salmonella choleraesuis (S. choleraesuis), Salmonella typhimurium (S. typhimurium), Escherichia coli (E. coli), and Brachyspira hyodysenteriae (B. hyodysenteriae). IFA and IHC results indicated that the monoclonal antibodies can be successfully used as primary antibodies to detect L. intracellularis in infected cells and in the crypt of the ileum from infected tissues of PE. Our findings suggested that the new monoclonal antibody specific against L. intracellularis will be useful for the evaluation of L. intracellularis infection in vivo and in vitro.Ning XiaoJiannan LiMinxue LiYuting HuHuixing LinHongjie FanHongjie FanFrontiers Media S.A.articleproliferative enteropathyLawsonia intracellularisOmp2 proteinmonoclonal antibodyindirect immunofluorescence assayimmunocytochemistryVeterinary medicineSF600-1100ENFrontiers in Veterinary Science, Vol 8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	proliferative enteropathy Lawsonia intracellularis Omp2 protein monoclonal antibody indirect immunofluorescence assay immunocytochemistry Veterinary medicine SF600-1100
spellingShingle	proliferative enteropathy Lawsonia intracellularis Omp2 protein monoclonal antibody indirect immunofluorescence assay immunocytochemistry Veterinary medicine SF600-1100 Ning Xiao Jiannan Li Minxue Li Yuting Hu Huixing Lin Hongjie Fan Hongjie Fan Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay
description	Proliferative enteropathy (PE) is an infectious enteric disease caused by Lawsonia intracellularis (L. intracellularis) and is endemic in pig herds worldwide. However, a L. intracellularis-specific monoclonal antibody plays an important role in the evaluation of L. intracellularis infection in vitro. Therefore, the objective of this study was to produce and identify the characteristics of a new monoclonal antibody against the outer membrane protein (Omp2) of L. intracellularis and apply it in an indirect immunofluorescence assay (IFA) and immunocytochemistry (IHC). The results indicated that three highly specific monoclonal antibodies against the Omp2 protein (4D9, 3G2, and 7G5) of L. intracellularis were obtained by using purified Omp2 as an immunogen, the titers of ascitic fluids of 4D9, 3G2, and 7G5 cells were 1:2,048,000, 1:512,000, and 1:256,000, respectively. IFA analysis showed that the 4D9, 3G2, and 7G5 have no cross-reactivity with other enteric bacteria commonly found in the ilea of pigs or closely related to L. intracellularis, such as Desulfovibrio, Bilophila wadsworthia (B. wadsworthia), Salmonella choleraesuis (S. choleraesuis), Salmonella typhimurium (S. typhimurium), Escherichia coli (E. coli), and Brachyspira hyodysenteriae (B. hyodysenteriae). IFA and IHC results indicated that the monoclonal antibodies can be successfully used as primary antibodies to detect L. intracellularis in infected cells and in the crypt of the ileum from infected tissues of PE. Our findings suggested that the new monoclonal antibody specific against L. intracellularis will be useful for the evaluation of L. intracellularis infection in vivo and in vitro.
format	article
author	Ning Xiao Jiannan Li Minxue Li Yuting Hu Huixing Lin Hongjie Fan Hongjie Fan
author_facet	Ning Xiao Jiannan Li Minxue Li Yuting Hu Huixing Lin Hongjie Fan Hongjie Fan
author_sort	Ning Xiao
title	Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay
title_short	Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay
title_full	Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay
title_fullStr	Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay
title_full_unstemmed	Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay
title_sort	preparation and characterization of a new monoclonal antibody specific against lawsonia intracellularis and its application in indirect immunofluorescence and immunocytochemistry assay
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/7f326a1807b544d4b5ec8d19701e92d4
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Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay

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