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Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection

The CD69 molecule, as an early activation marker of lymphocytes, is often used to assess the activation of cellular immunity. However, for pigs, an anti-pig CD69 antibody is not yet available for this purpose after infection or vaccination. In this study, a monoclonal antibody (mAb) against pig CD69...

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Autores principales: Tian, Yunfei, Hao, Yuxin, Dong, Maoli, Li, Shuai, Wang, Dongyue, Jiang, Fei, Wang, Qingqing, Hao, Xiaoli, Yang, Yi, Chen, Nanhua, Zhu, Jianzhong, Guo, Junqing, Wu, Jiajun, Shang, Shaobin, Zhou, Jiyong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9231377/
https://www.ncbi.nlm.nih.gov/pubmed/35746813
http://dx.doi.org/10.3390/v14061343
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author Tian, Yunfei
Hao, Yuxin
Dong, Maoli
Li, Shuai
Wang, Dongyue
Jiang, Fei
Wang, Qingqing
Hao, Xiaoli
Yang, Yi
Chen, Nanhua
Zhu, Jianzhong
Guo, Junqing
Wu, Jiajun
Shang, Shaobin
Zhou, Jiyong
author_facet Tian, Yunfei
Hao, Yuxin
Dong, Maoli
Li, Shuai
Wang, Dongyue
Jiang, Fei
Wang, Qingqing
Hao, Xiaoli
Yang, Yi
Chen, Nanhua
Zhu, Jianzhong
Guo, Junqing
Wu, Jiajun
Shang, Shaobin
Zhou, Jiyong
author_sort Tian, Yunfei
collection PubMed
description The CD69 molecule, as an early activation marker of lymphocytes, is often used to assess the activation of cellular immunity. However, for pigs, an anti-pig CD69 antibody is not yet available for this purpose after infection or vaccination. In this study, a monoclonal antibody (mAb) against pig CD69 was produced by peptide immunization and hybridoma technique. One mAb (5F12) showed good reactivity with pig CD69 that was expressed in transfected-HEK-293T cells and on mitogen-activated porcine peripheral blood mononuclear cells (PBMCs) by indirect immunofluorescence assay and flow cytometry. This mAb did not cross-react with activated lymphocytes from mouse, bovine, and chicken. Epitope mapping showed that the epitope recognized by this mAb was located at amino acid residues 147–161 of pig CD69. By conjugating with fluorochrome, this mAb was used to detect the early activation of lymphocytes in PRRSV- and ASFV-infected pigs by flow cytometry. The results showed that PRRSV infection induced the dominant activation of CD4 T cells in mediastinal lymph nodes and CD8 T cells in the spleen at 14 days post-infection, in terms of CD69 expression. In an experiment on ASFV infection, we found that ASFV infection resulted in the early activation of NK cells, B cells, and distinct T cell subsets with variable magnitude in PBMCs, spleen, and submandibular lymph nodes. Our study revealed an early event of lymphocyte and T cell activation after PRRSV and ASFV infections and provides an important immunological tool for the in-depth analysis of cellular immune response in pigs after infection or vaccination.
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spelling pubmed-92313772022-06-25 Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection Tian, Yunfei Hao, Yuxin Dong, Maoli Li, Shuai Wang, Dongyue Jiang, Fei Wang, Qingqing Hao, Xiaoli Yang, Yi Chen, Nanhua Zhu, Jianzhong Guo, Junqing Wu, Jiajun Shang, Shaobin Zhou, Jiyong Viruses Article The CD69 molecule, as an early activation marker of lymphocytes, is often used to assess the activation of cellular immunity. However, for pigs, an anti-pig CD69 antibody is not yet available for this purpose after infection or vaccination. In this study, a monoclonal antibody (mAb) against pig CD69 was produced by peptide immunization and hybridoma technique. One mAb (5F12) showed good reactivity with pig CD69 that was expressed in transfected-HEK-293T cells and on mitogen-activated porcine peripheral blood mononuclear cells (PBMCs) by indirect immunofluorescence assay and flow cytometry. This mAb did not cross-react with activated lymphocytes from mouse, bovine, and chicken. Epitope mapping showed that the epitope recognized by this mAb was located at amino acid residues 147–161 of pig CD69. By conjugating with fluorochrome, this mAb was used to detect the early activation of lymphocytes in PRRSV- and ASFV-infected pigs by flow cytometry. The results showed that PRRSV infection induced the dominant activation of CD4 T cells in mediastinal lymph nodes and CD8 T cells in the spleen at 14 days post-infection, in terms of CD69 expression. In an experiment on ASFV infection, we found that ASFV infection resulted in the early activation of NK cells, B cells, and distinct T cell subsets with variable magnitude in PBMCs, spleen, and submandibular lymph nodes. Our study revealed an early event of lymphocyte and T cell activation after PRRSV and ASFV infections and provides an important immunological tool for the in-depth analysis of cellular immune response in pigs after infection or vaccination. MDPI 2022-06-20 /pmc/articles/PMC9231377/ /pubmed/35746813 http://dx.doi.org/10.3390/v14061343 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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/).
spellingShingle Article
Tian, Yunfei
Hao, Yuxin
Dong, Maoli
Li, Shuai
Wang, Dongyue
Jiang, Fei
Wang, Qingqing
Hao, Xiaoli
Yang, Yi
Chen, Nanhua
Zhu, Jianzhong
Guo, Junqing
Wu, Jiajun
Shang, Shaobin
Zhou, Jiyong
Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection
title Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection
title_full Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection
title_fullStr Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection
title_full_unstemmed Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection
title_short Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection
title_sort development of a monoclonal antibody to pig cd69 reveals early activation of t cells in pig after prrsv and asfv infection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9231377/
https://www.ncbi.nlm.nih.gov/pubmed/35746813
http://dx.doi.org/10.3390/v14061343
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