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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MDPI AG
2022-06-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/6/1343 |
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| author | Yunfei Tian Yuxin Hao Maoli Dong Shuai Li Dongyue Wang Fei Jiang Qingqing Wang Xiaoli Hao Yi Yang Nanhua Chen Jianzhong Zhu Junqing Guo Jiajun Wu Shaobin Shang Jiyong Zhou |
| author_facet | Yunfei Tian Yuxin Hao Maoli Dong Shuai Li Dongyue Wang Fei Jiang Qingqing Wang Xiaoli Hao Yi Yang Nanhua Chen Jianzhong Zhu Junqing Guo Jiajun Wu Shaobin Shang Jiyong Zhou |
| author_sort | Yunfei Tian |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T22:13:39Z |
| format | Article |
| id | doaj.art-b617529ca9d241ff8385ce257aad0ae9 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T22:13:39Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-b617529ca9d241ff8385ce257aad0ae92023-11-23T19:27:47ZengMDPI AGViruses1999-49152022-06-01146134310.3390/v14061343Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV InfectionYunfei Tian0Yuxin Hao1Maoli Dong2Shuai Li3Dongyue Wang4Fei Jiang5Qingqing Wang6Xiaoli Hao7Yi Yang8Nanhua Chen9Jianzhong Zhu10Junqing Guo11Jiajun Wu12Shaobin Shang13Jiyong Zhou14Institute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaThe Biosafety High-Level Laboratory Management Office, China Animal Disease Control Center, Beijing 102609, ChinaInstitute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaInstitute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaThe Biosafety High-Level Laboratory Management Office, China Animal Disease Control Center, Beijing 102609, ChinaThe Biosafety High-Level Laboratory Management Office, China Animal Disease Control Center, Beijing 102609, ChinaThe Biosafety High-Level Laboratory Management Office, China Animal Disease Control Center, Beijing 102609, ChinaInstitute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaInstitute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaInstitute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaInstitute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaKey Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450008, ChinaThe Biosafety High-Level Laboratory Management Office, China Animal Disease Control Center, Beijing 102609, ChinaInstitute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaCollege of Animal Science, Zhejiang University, Hangzhou 310058, ChinaThe 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.https://www.mdpi.com/1999-4915/14/6/1343porcine CD69monoclonal antibodyT cell activationPRRSVASFV |
| spellingShingle | Yunfei Tian Yuxin Hao Maoli Dong Shuai Li Dongyue Wang Fei Jiang Qingqing Wang Xiaoli Hao Yi Yang Nanhua Chen Jianzhong Zhu Junqing Guo Jiajun Wu Shaobin Shang Jiyong Zhou Development of a Monoclonal Antibody to Pig CD69 Reveals Early Activation of T Cells in Pig after PRRSV and ASFV Infection Viruses porcine CD69 monoclonal antibody T cell activation PRRSV ASFV |
| 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 | porcine CD69 monoclonal antibody T cell activation PRRSV ASFV |
| url | https://www.mdpi.com/1999-4915/14/6/1343 |
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