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Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus

Cytotoxic T lymphocytes (CTL) are key players of the adaptive immune system that target tumors and infected cells. A central step to that is the formation of a cell–cell contact zone between the CTL and its target called an immune synapse (IS). Here, we investigate the influence of the initial T cel...

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Autores principales: Estl, Michael, Blatt, Pascal, Li, Xuemei, Becherer, Ute, Chang, Hsin-Fang, Rettig, Jens, Pattu, Varsha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7177831/
https://www.ncbi.nlm.nih.gov/pubmed/32252488
http://dx.doi.org/10.3390/ijms21072475
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author Estl, Michael
Blatt, Pascal
Li, Xuemei
Becherer, Ute
Chang, Hsin-Fang
Rettig, Jens
Pattu, Varsha
author_facet Estl, Michael
Blatt, Pascal
Li, Xuemei
Becherer, Ute
Chang, Hsin-Fang
Rettig, Jens
Pattu, Varsha
author_sort Estl, Michael
collection PubMed
description Cytotoxic T lymphocytes (CTL) are key players of the adaptive immune system that target tumors and infected cells. A central step to that is the formation of a cell–cell contact zone between the CTL and its target called an immune synapse (IS). Here, we investigate the influence of the initial T cell receptor (TCR) trigger of a cytolytic IS on the distinct steps leading to cytotoxic granule (CG) exocytosis. We stimulated primary CTLs from mouse using lipid bilayers with varying anti-CD3 but constant ICAM concentrations. We fluorescently labeled molecular markers of distinct IS zones such as actin, CD3, granzyme B, and Synaptobrevin2 in CTLs and imaged cytolytic IS formation by total internal reflection fluorescence microscopy (TIRFM). We found that an intermediate anti-CD3 concentration of 10 µg/mL induces the fastest adhesion of CTLs to the bilayers and results in maximal CG fusion efficiency. The latency of actin ring formation, dwell time, and maximum surface area at the IS exhibit different dependencies on the stimulatory anti-CD3 concentrations. The number and surface area of CD3 clusters at the IS seem to show a different dependency to the TCR trigger when compared to their dwell time. Finally, the mode of full CG exocytosis appears to be independent of the TCR trigger.
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spelling pubmed-71778312020-04-28 Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus Estl, Michael Blatt, Pascal Li, Xuemei Becherer, Ute Chang, Hsin-Fang Rettig, Jens Pattu, Varsha Int J Mol Sci Article Cytotoxic T lymphocytes (CTL) are key players of the adaptive immune system that target tumors and infected cells. A central step to that is the formation of a cell–cell contact zone between the CTL and its target called an immune synapse (IS). Here, we investigate the influence of the initial T cell receptor (TCR) trigger of a cytolytic IS on the distinct steps leading to cytotoxic granule (CG) exocytosis. We stimulated primary CTLs from mouse using lipid bilayers with varying anti-CD3 but constant ICAM concentrations. We fluorescently labeled molecular markers of distinct IS zones such as actin, CD3, granzyme B, and Synaptobrevin2 in CTLs and imaged cytolytic IS formation by total internal reflection fluorescence microscopy (TIRFM). We found that an intermediate anti-CD3 concentration of 10 µg/mL induces the fastest adhesion of CTLs to the bilayers and results in maximal CG fusion efficiency. The latency of actin ring formation, dwell time, and maximum surface area at the IS exhibit different dependencies on the stimulatory anti-CD3 concentrations. The number and surface area of CD3 clusters at the IS seem to show a different dependency to the TCR trigger when compared to their dwell time. Finally, the mode of full CG exocytosis appears to be independent of the TCR trigger. MDPI 2020-04-02 /pmc/articles/PMC7177831/ /pubmed/32252488 http://dx.doi.org/10.3390/ijms21072475 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Estl, Michael
Blatt, Pascal
Li, Xuemei
Becherer, Ute
Chang, Hsin-Fang
Rettig, Jens
Pattu, Varsha
Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus
title Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus
title_full Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus
title_fullStr Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus
title_full_unstemmed Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus
title_short Various Stages of Immune Synapse Formation Are Differently Dependent on the Strength of the TCR Stimulus
title_sort various stages of immune synapse formation are differently dependent on the strength of the tcr stimulus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7177831/
https://www.ncbi.nlm.nih.gov/pubmed/32252488
http://dx.doi.org/10.3390/ijms21072475
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