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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...

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Autores principales: Zechmann, Maria, Reese, Sven, Göbel, Thomas W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
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.
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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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