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Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8(+) αβ and γδ T Lymphocytes with Different Kinetics
During a search for immunomodulatory receptors in the chicken genome, we identified a previously cloned chicken sequence as CRTAM homologue by its overall identity and several conserved sequence features. For further characterization, we generated a CRTAM specific mab. No staining was detectable in...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2013
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3858274/ https://www.ncbi.nlm.nih.gov/pubmed/24339981 http://dx.doi.org/10.1371/journal.pone.0081942 |
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author | Zechmann, Maria Reese, Sven Göbel, Thomas W. |
author_facet | Zechmann, Maria Reese, Sven Göbel, Thomas W. |
author_sort | Zechmann, Maria |
collection | PubMed |
description | During a search for immunomodulatory receptors in the chicken genome, we identified a previously cloned chicken sequence as CRTAM homologue by its overall identity and several conserved sequence features. For further characterization, we generated a CRTAM specific mab. No staining was detectable in freshly isolated cell preparations from thymus, bursa, caecal tonsils, spleen, blood and intestine. Activation of splenocytes with recombinant IL-2 increased rapid CRTAM expression within a 2 h period on about 30% of the cells. These CRTAM(+) cells were identified as CD8(+) γδ T lymphocytes. In contrast, CRTAM expression could not be stimulated on PBL with IL-2, even within a 48 h stimulation period. As a second means of activation, T cell receptor (TCR) crosslinking using an anti-αβ-TCR induced CRTAM on both PBL and splenocytes. While CRTAM expression was again rapidly upregulated on splenocytes within 2 h, it took 48 h to reach maximum levels of CRTAM expression in PBL. Strikingly, albeit the stimulation of splenocytes was performed with anti-αβ-TCR, CRTAM expression after 2 h was mainly restricted to CD8(+) γδ T lymphocytes, however, the longer anti-TCR stimulation of peripheral blood lymphocytes (PBL) resulted in CRTAM expression on αβ T lymphocytes. In order to characterize the potential ligand we cloned and expressed chicken Necl-2, a member of the nectin and nectin-like family which is highly homologous to its mammalian counterpart. Three independent assays including a reporter assay, staining with a CRTAM-Ig fusion protein and a cell conjugate assay confirmed the interaction of CRTAM with Necl-2 which could also be blocked by a soluble CRTAM-Ig fusion protein or a CRTAM specific mab. These results suggest that chicken CRTAM represents an early activation antigen on CD8(+) T cells which binds to Necl-2 and is upregulated with distinct kinetics on αβ versus γδ T lymphocytes. |
format | Online Article Text |
id | pubmed-3858274 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-38582742013-12-11 Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8(+) αβ and γδ T Lymphocytes with Different Kinetics Zechmann, Maria Reese, Sven Göbel, Thomas W. PLoS One Research Article During a search for immunomodulatory receptors in the chicken genome, we identified a previously cloned chicken sequence as CRTAM homologue by its overall identity and several conserved sequence features. For further characterization, we generated a CRTAM specific mab. No staining was detectable in freshly isolated cell preparations from thymus, bursa, caecal tonsils, spleen, blood and intestine. Activation of splenocytes with recombinant IL-2 increased rapid CRTAM expression within a 2 h period on about 30% of the cells. These CRTAM(+) cells were identified as CD8(+) γδ T lymphocytes. In contrast, CRTAM expression could not be stimulated on PBL with IL-2, even within a 48 h stimulation period. As a second means of activation, T cell receptor (TCR) crosslinking using an anti-αβ-TCR induced CRTAM on both PBL and splenocytes. While CRTAM expression was again rapidly upregulated on splenocytes within 2 h, it took 48 h to reach maximum levels of CRTAM expression in PBL. Strikingly, albeit the stimulation of splenocytes was performed with anti-αβ-TCR, CRTAM expression after 2 h was mainly restricted to CD8(+) γδ T lymphocytes, however, the longer anti-TCR stimulation of peripheral blood lymphocytes (PBL) resulted in CRTAM expression on αβ T lymphocytes. In order to characterize the potential ligand we cloned and expressed chicken Necl-2, a member of the nectin and nectin-like family which is highly homologous to its mammalian counterpart. Three independent assays including a reporter assay, staining with a CRTAM-Ig fusion protein and a cell conjugate assay confirmed the interaction of CRTAM with Necl-2 which could also be blocked by a soluble CRTAM-Ig fusion protein or a CRTAM specific mab. These results suggest that chicken CRTAM represents an early activation antigen on CD8(+) T cells which binds to Necl-2 and is upregulated with distinct kinetics on αβ versus γδ T lymphocytes. Public Library of Science 2013-12-10 /pmc/articles/PMC3858274/ /pubmed/24339981 http://dx.doi.org/10.1371/journal.pone.0081942 Text en © 2013 Zechmann et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Zechmann, Maria Reese, Sven Göbel, Thomas W. Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8(+) αβ and γδ T Lymphocytes with Different Kinetics |
title | Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8(+) αβ and γδ T Lymphocytes with Different Kinetics |
title_full | Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8(+) αβ and γδ T Lymphocytes with Different Kinetics |
title_fullStr | Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8(+) αβ and γδ T Lymphocytes with Different Kinetics |
title_full_unstemmed | Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8(+) αβ and γδ T Lymphocytes with Different Kinetics |
title_short | Chicken CRTAM Binds Nectin-Like 2 Ligand and Is Upregulated on CD8(+) αβ and γδ T Lymphocytes with Different Kinetics |
title_sort | chicken crtam binds nectin-like 2 ligand and is upregulated on cd8(+) αβ and γδ t lymphocytes with different kinetics |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3858274/ https://www.ncbi.nlm.nih.gov/pubmed/24339981 http://dx.doi.org/10.1371/journal.pone.0081942 |
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