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:	Maria Zechmann, Sven Reese, Thomas W Göbel
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e603f386c3c944d7beee1b2b120ba22c
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spelling	oai:doaj.org-article:e603f386c3c944d7beee1b2b120ba22c2021-11-18T08:42:42ZChicken CRTAM binds nectin-like 2 ligand and is upregulated on CD8+ αβ and γδ T lymphocytes with different kinetics.1932-620310.1371/journal.pone.0081942https://doaj.org/article/e603f386c3c944d7beee1b2b120ba22c2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24339981/?tool=EBIhttps://doaj.org/toc/1932-6203During 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.Maria ZechmannSven ReeseThomas W GöbelPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 12, p e81942 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maria Zechmann Sven Reese Thomas W Göbel Chicken CRTAM binds nectin-like 2 ligand and is upregulated on CD8+ αβ and γδ T lymphocytes with different kinetics.
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	article
author	Maria Zechmann Sven Reese Thomas W Göbel
author_facet	Maria Zechmann Sven Reese Thomas W Göbel
author_sort	Maria Zechmann
title	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_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_sort	chicken crtam binds nectin-like 2 ligand and is upregulated on cd8+ αβ and γδ t lymphocytes with different kinetics.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/e603f386c3c944d7beee1b2b120ba22c
work_keys_str_mv	AT mariazechmann chickencrtambindsnectinlike2ligandandisupregulatedoncd8abandgdtlymphocyteswithdifferentkinetics AT svenreese chickencrtambindsnectinlike2ligandandisupregulatedoncd8abandgdtlymphocyteswithdifferentkinetics AT thomaswgobel chickencrtambindsnectinlike2ligandandisupregulatedoncd8abandgdtlymphocyteswithdifferentkinetics
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Chicken CRTAM binds nectin-like 2 ligand and is upregulated on CD8+ αβ and γδ T lymphocytes with different kinetics.

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