The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.

Various aspects of the in vitro culture conditions can impact the functional response of immune cells. For example, it was shown that a Ca2+ concentration of at least 1.5 mM during in vitro stimulation is needed for optimal cytokine production by conventional αβ T cells. Here we extend these finding...

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Main Authors: Yusuf Cem Eskiocak, Zeynep Ozge Ayyildiz, Sinem Gunalp, Asli Korkmaz, Derya Goksu Helvaci, Yavuz Dogan, Duygu Sag, Gerhard Wingender
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2023-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0282037
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author Yusuf Cem Eskiocak
Zeynep Ozge Ayyildiz
Sinem Gunalp
Asli Korkmaz
Derya Goksu Helvaci
Yavuz Dogan
Duygu Sag
Gerhard Wingender
author_facet Yusuf Cem Eskiocak
Zeynep Ozge Ayyildiz
Sinem Gunalp
Asli Korkmaz
Derya Goksu Helvaci
Yavuz Dogan
Duygu Sag
Gerhard Wingender
author_sort Yusuf Cem Eskiocak
collection DOAJ
description Various aspects of the in vitro culture conditions can impact the functional response of immune cells. For example, it was shown that a Ca2+ concentration of at least 1.5 mM during in vitro stimulation is needed for optimal cytokine production by conventional αβ T cells. Here we extend these findings by showing that also unconventional T cells (invariant Natural Killer T cells, mucosal-associated invariant T cells, γδ T cells), as well as B cells, show an increased cytokine response following in vitro stimulation in the presence of elevated Ca2+ concentrations. This effect appeared more pronounced with mouse than with human lymphoid cells and did not influence their survival. A similarly increased cytokine response due to elevated Ca2+ levels was observed with primary human monocytes. In contrast, primary human monocyte-derived macrophages, either unpolarized (M0) or polarized into M1 or M2 macrophages, displayed increased cell death in the presence of elevated Ca2+ concentrations. Furthermore, elevated Ca2+ concentrations promoted phenotypic M1 differentiation by increasing M1 markers on M1 and M2 macrophages and decreasing M2 markers on M2 macrophages. However, the cytokine production of macrophages, again in contrast to the lymphoid cells, was unaltered by the Ca2+ concentration. In summary, our data demonstrate that the Ca2+ concentration during in vitro cultures is an important variable to be considered for functional experiments and that elevated Ca2+ levels can boost cytokine production by both mouse and human lymphoid cells.
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spelling doaj.art-74ddccde7b06412bbe4a9e10248401d72023-04-12T05:33:10ZengPublic Library of Science (PLoS)PLoS ONE1932-62032023-01-01182e028203710.1371/journal.pone.0282037The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.Yusuf Cem EskiocakZeynep Ozge AyyildizSinem GunalpAsli KorkmazDerya Goksu HelvaciYavuz DoganDuygu SagGerhard WingenderVarious aspects of the in vitro culture conditions can impact the functional response of immune cells. For example, it was shown that a Ca2+ concentration of at least 1.5 mM during in vitro stimulation is needed for optimal cytokine production by conventional αβ T cells. Here we extend these findings by showing that also unconventional T cells (invariant Natural Killer T cells, mucosal-associated invariant T cells, γδ T cells), as well as B cells, show an increased cytokine response following in vitro stimulation in the presence of elevated Ca2+ concentrations. This effect appeared more pronounced with mouse than with human lymphoid cells and did not influence their survival. A similarly increased cytokine response due to elevated Ca2+ levels was observed with primary human monocytes. In contrast, primary human monocyte-derived macrophages, either unpolarized (M0) or polarized into M1 or M2 macrophages, displayed increased cell death in the presence of elevated Ca2+ concentrations. Furthermore, elevated Ca2+ concentrations promoted phenotypic M1 differentiation by increasing M1 markers on M1 and M2 macrophages and decreasing M2 markers on M2 macrophages. However, the cytokine production of macrophages, again in contrast to the lymphoid cells, was unaltered by the Ca2+ concentration. In summary, our data demonstrate that the Ca2+ concentration during in vitro cultures is an important variable to be considered for functional experiments and that elevated Ca2+ levels can boost cytokine production by both mouse and human lymphoid cells.https://doi.org/10.1371/journal.pone.0282037
spellingShingle Yusuf Cem Eskiocak
Zeynep Ozge Ayyildiz
Sinem Gunalp
Asli Korkmaz
Derya Goksu Helvaci
Yavuz Dogan
Duygu Sag
Gerhard Wingender
The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.
PLoS ONE
title The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.
title_full The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.
title_fullStr The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.
title_full_unstemmed The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.
title_short The Ca2+ concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro.
title_sort ca2 concentration impacts the cytokine production of mouse and human lymphoid cells and the polarization of human macrophages in vitro
url https://doi.org/10.1371/journal.pone.0282037
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