Preparation and Characterization of a New Monoclonal Antibody Specific Against Lawsonia intracellularis and Its Application in Indirect Immunofluorescence and Immunocytochemistry Assay.
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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DOI：
10.3389/fvets.2021.753610
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发表时间：
2021
影响因子：
3.2
通讯作者：
Fan H
中科院分区：
文献类型：
--
作者：
Xiao N;Li J;Li M;Hu Y;Lin H;Fan H
增殖性肠病（PE）是由胞内Lawsonia intracellularis （L.胞内Lawsonia intracellularis）引起的一种传染性肠道疾病，在世界范围内的猪群中流行。然而，胞内乳杆菌特异性单克隆抗体在体外评价胞内乳杆菌感染中起着重要作用。因此，本研究的目的是制备和鉴定一种新的抗胞内乳杆菌外膜蛋白（Omp2）单克隆抗体，并将其应用于间接免疫荧光法（IFA）和免疫细胞化学（IHC）。结果表明，以纯化的Omp2为免疫原，可获得3种高特异性的单克隆抗体（4D9、3G2和7G5）， 4D9、3G2和7G5细胞的腹水滴度分别为1:20,48,000、1:51,12,000和1:25,6,000。IFA分析表明，4D9、3G2和7G5与猪回肠中常见或与胞内乳杆菌密切相关的其他肠道细菌如Desulfovibrio、Bilophila wadsworthia （B. wadsworthia）、沙门氏菌choleraesuis （S. choleraesuis）、鼠伤寒沙门氏菌（S. typhimurium）、大肠杆菌（E. coli）、水痢短螺旋体（B. hyodysenteriae）无交叉反应性。IFA和IHC结果表明，该单克隆抗体可成功地作为一抗检测PE感染组织中感染细胞和回肠隐窝中的胞内乳杆菌。本研究结果提示，该单克隆抗体可用于胞内乳杆菌体内和体外感染评价。
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.
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影响因子：
3.7
作者：
Ti J;Li Z;Li X;Lu Y;Diao Y;Li F
通讯作者：
Li F
影响因子：
2.4
作者：
Boye, M;Jensen, TK;Jorsal, SE
通讯作者：
Jorsal, SE
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3.3
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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1.5
作者：
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通讯作者：
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