Cytotoxic Activity of CD4 T Cells During the Early Stage of Autoimmune Neuroinflammation

Pradella, Fernando; Vo, Boldrini; Am, Marques; Gad, Morais; Francelin, Carolina; Rs, Cocenza; Vc, Lima; Bonora, Massimo; Ns, Brunetti; Bb, Campos; Esm, Fonseca; Rocha-Parise, Michelle; Crv, Stella; Damasceno, Alfredo; F, von Glehn; Alf, Longhini; Lmb, Santos; Farias, Alessandro S.

doi:10.1101/2020.03.10.985614

Cited by 5 publications

(6 citation statements)

References 57 publications

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“…Interestingly, we observed that the expression (raw counts) of CD28 mRNA was generally low, especially in cells with high expression of GZMB in Th17-polarized cells in coculture with OLs by single-cell RNA sequencing ( Supplementary Figure 9B ). Granzyme B levels are elevated in the CSF of MS patients ( 42 ) and CD4 + T cells from MS patients secrete more granzyme B than healthy controls in the relapsing–remitting ( 85 ) and progressive forms ( 86 ). In addition, pro-inflammatory CD4 + T cells secreting granzyme B can disrupt the blood–brain barrier ( 87 ) and cause glial fibrillary acid protein fragmentation in human primary astrocytes in vitro ( 22 ).…”

Section: Discussionmentioning

confidence: 99%

Contact-Dependent Granzyme B-Mediated Cytotoxicity of Th17-Polarized Cells Toward Human Oligodendrocytes

et al. 2022

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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 natural granzyme B blocker (serpina3N) improved survival of MO3.13 cells upon coculture with Th17 cells. In conclusion, we showed that human Th17-polarized cells form biologically significant contacts with human OLs and exert direct toxicity by releasing granzyme B.

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Section: Discussionmentioning

confidence: 99%

Contact-Dependent Granzyme B-Mediated Cytotoxicity of Th17-Polarized Cells Toward Human Oligodendrocytes

et al. 2022

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“…Here, we provide evidence of cytotoxic behavior derived from CD3+CD20+ T cell during progressive MS. Recently, we demonstrated that, despite not known as a classical cytotoxic subset, the ectopic expression of GzmB by CD4+ T cells is increased in the peripheral blood from RRMS patients particularly during Natalizumab treatment when compared to other MS therapies, untreated MS patients and healthy donors (Pradella et al, 2020). Importantly, classical CD8+GzmB+ and ectopic CD4+GzmB+ T cells were shown into CNS-lesions from fulminant MS relapses after Natalizumab discontinuation (Larochelle et al, 2017;Serafini et al, 2017).…”

Section: Discussionmentioning

confidence: 92%

“…Perforin-and serine protease granzyme-B (GzmB)-derived circulating CD8+ and CD4+ T lymphocytes from MS patients were previously identified (Broux et al, 2015;Cencioni et al, 2017;Fujii et al, 2016;Pradella et al, 2020). However, this cytotoxic behavior was not yet investigated in CD3+CD20+ T cells from MS patients.…”

Section: Expression Of Perforin and Serine-protease Granzyme-b (Gzmb)...mentioning

confidence: 99%

Cytotoxic profile of CD3+CD20+ T cells in progressive multiple sclerosis

Boldrini

Quintiliano

Silva

et al. 2021

Multiple Sclerosis and Related Disorders

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“…While these findings supported the idea that CD4 + T cells responding to Myhc-α 334-352 may possess the characteristics of cytotoxic CD4 + T cells, we sought to further validate them by testing the expression of other newly described markers of CD4 + CTLs. These included surface expression of NKG2A [50], NKG2D [51], and CRTAM [52] and intracellular expression of EOMES [53][54][55]. In a flow cytometric analysis, we compared the expression of CD107a with that of NKG2A, NKG2D, CRTAM, and EOMES, expecting to find that their detection aligned with CD107a expression.…”

Section: Resultsmentioning

confidence: 99%

Investigation into Cardiac Myhc-α 334–352-Specific TCR Transgenic Mice Reveals a Role for Cytotoxic CD4 T Cells in the Development of Cardiac Autoimmunity

Sur,

Rasquinha,

Mone

et al. 2024

Cells

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Myocarditis is one of the major causes of heart failure in children and young adults and can lead to dilated cardiomyopathy. Lymphocytic myocarditis could result from autoreactive CD4+ and CD8+ T cells, but defining antigen specificity in disease pathogenesis is challenging. To address this issue, we generated T cell receptor (TCR) transgenic (Tg) C57BL/6J mice specific to cardiac myosin heavy chain (Myhc)-α 334–352 and found that Myhc-α-specific TCRs were expressed in both CD4+ and CD8+ T cells. To investigate if the phenotype is more pronounced in a myocarditis-susceptible genetic background, we backcrossed with A/J mice. At the fourth generation of backcrossing, we observed that Tg T cells from naïve mice responded to Myhc-α 334–352, as evaluated by proliferation assay and carboxyfluorescein succinimidyl ester staining. The T cell responses included significant production of mainly pro-inflammatory cytokines, namely interferon (IFN)-γ, interleukin-17, and granulocyte macrophage-colony stimulating factor. While the naïve Tg mice had isolated myocardial lesions, immunization with Myhc-α 334–352 led to mild myocarditis, suggesting that further backcrossing to increase the percentage of A/J genome close to 99.99% might show a more severe disease phenotype. Further investigations led us to note that CD4+ T cells displayed the phenotype of cytotoxic T cells (CTLs) akin to those of conventional CD8+ CTLs, as determined by the expression of CD107a, IFN-γ, granzyme B natural killer cell receptor (NKG)2A, NKG2D, cytotoxic and regulatory T cell molecules, and eomesodermin. Taken together, the transgenic system described in this report may be a helpful tool to distinguish the roles of cytotoxic cardiac antigen-specific CD4+ T cells vs. those of CD8+ T cells in the pathogenesis of myocarditis.

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Cytotoxic Activity of CD4 T Cells During the Early Stage of Autoimmune Neuroinflammation

Cited by 5 publications

References 57 publications

Contact-Dependent Granzyme B-Mediated Cytotoxicity of Th17-Polarized Cells Toward Human Oligodendrocytes

Contact-Dependent Granzyme B-Mediated Cytotoxicity of Th17-Polarized Cells Toward Human Oligodendrocytes

Cytotoxic profile of CD3+CD20+ T cells in progressive multiple sclerosis

Investigation into Cardiac Myhc-α 334–352-Specific TCR Transgenic Mice Reveals a Role for Cytotoxic CD4 T Cells in the Development of Cardiac Autoimmunity

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