Contact-Dependent Granzyme B-Mediated Cytotoxicity of Th17-Polarized Cells Toward Human Oligodendrocytes
In: Front Immunol, 2022
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Zugriff:
Multiple sclerosis (MS) is characterized by the loss of myelin and of myelin-producing oligodendrocytes (OLs) in the central nervous system (CNS). Pro-inflammatory CD4(+) Th17 cells are considered pathogenic in MS and are harmful to OLs. We investigated the mechanisms driving human CD4(+) T cell-mediated OL cell death. Using fluorescent and brightfield in vitro live imaging, we found that compared to Th2-polarized cells, Th17-polarized cells show greater interactions with primary human OLs and human oligodendrocytic cell line MO3.13, displaying longer duration of contact, lower mean speed, and higher rate of vesicle-like structure formation at the sites of contact. Using single-cell RNA sequencing, we assessed the transcriptomic profile of primary human OLs and Th17-polarized cells in direct contact or separated by an insert. We showed that upon close interaction, OLs upregulate the expression of mRNA coding for chemokines and antioxidant/anti-apoptotic molecules, while Th17-polarized cells upregulate the expression of mRNA coding for chemokines and pro-inflammatory cytokines such as IL-17A, IFN-γ, and granzyme B. We found that secretion of CCL3, CXCL10, IFN-γ, TNFα, and granzyme B is induced upon direct contact in cocultures of human Th17-polarized cells with human OLs. In addition, we validated by flow cytometry and immunofluorescence that granzyme B levels are upregulated in Th17-polarized compared to Th2-polarized cells and are even higher in Th17-polarized cells upon direct contact with OLs or MO3.13 cells compared to Th17-polarized cells separated from OLs by an insert. Moreover, granzyme B is detected in OLs and MO3.13 cells following direct contact with Th17-polarized cells, suggesting the release of granzyme B from Th17-polarized cells into OLs/MO3.13 cells. To confirm granzyme B–mediated cytotoxicity toward OLs, we showed that recombinant human granzyme B can induce OLs and MO3.13 cell death. Furthermore, pretreatment of Th17-polarized cells with a reversible granzyme B blocker (Ac-IEPD-CHO) or a ...
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Contact-Dependent Granzyme B-Mediated Cytotoxicity of Th17-Polarized Cells Toward Human Oligodendrocytes
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Autor/in / Beteiligte Person: | Jamann, Hélène ; Cui, Qiao-Ling ; Desu, Haritha L. ; Pernin, Florian ; Tastet, Olivier ; Halaweh, Alexandre ; Farzam-kia, Negar ; Mamane, Victoria Hannah ; Ouédraogo, Oumarou ; Cleret-Buhot, Aurélie ; Daigneault, Audrey ; Balthazard, Renaud ; Klement, Wendy ; Lemaître, Florent ; Arbour, Nathalie ; Antel, Jack ; Stratton, Jo Anne ; Larochelle, Catherine |
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Zeitschrift: | Front Immunol, 2022 |
Veröffentlichung: | Frontiers Media S.A., 2022 |
Medientyp: | academicJournal |
DOI: | 10.3389/fimmu.2022.850616 |
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