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The Macrophage Galactose-Type C-Type Lectin (MGL) Modulates Regulatory T Cell Functions.

Zizzari IG, Martufi P, Battisti F, Rahimi H, Caponnetto S, Bellati F, Nuti M, Rughetti A, Napoletano C - PLoS ONE (2015)

Bottom Line: In this study, we demonstrate that Treg immunosuppressive activity can be modulated by the cross-linking between the CD45RA expressed by Tregs and the C-type lectin MGL.This effect can be attributed to changes in CD45RA and TCR signalling through the inhibition of Lck and inactivation of Zap-70, an increase in the Foxp3 methylation status and, ultimately, the reduced production of suppressive cytokines.These results indicate a role of MGL as an immunomodulator within the tumour microenvironment interfering with Treg functions, suggesting its possible use in the design of anticancer vaccines.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medicine, "Sapienza" University, Rome, Italy.

ABSTRACT
Regulatory T cells (Tregs) are physiologically designed to prevent autoimmune disease and maintain self-tolerance. In tumour microenvironments, their presence is related to a poor prognosis, and they influence the therapeutic outcome due to their capacity to suppress the immune response by cell-cell contact and to release immunosuppressive cytokines. In this study, we demonstrate that Treg immunosuppressive activity can be modulated by the cross-linking between the CD45RA expressed by Tregs and the C-type lectin MGL. This specific interaction strongly decreases the immunosuppressive activity of Tregs, restoring the proliferative capacity of co-cultured T lymphocytes. This effect can be attributed to changes in CD45RA and TCR signalling through the inhibition of Lck and inactivation of Zap-70, an increase in the Foxp3 methylation status and, ultimately, the reduced production of suppressive cytokines. These results indicate a role of MGL as an immunomodulator within the tumour microenvironment interfering with Treg functions, suggesting its possible use in the design of anticancer vaccines.

No MeSH data available.


Related in: MedlinePlus

MGL-CD45RA interaction reduces immunosuppressive cytokine production.(A) Real-time PCR analysis of T-bet, RORγT and GATA3 genes in Tregs alone (grey columns) or after treatment with rhMGL-Fc (black columns). Relative gene expression was normalized with B2M and RLP0 gene expression levels. The results correspond to the mean value ± SD of three donors. (B) Intracellular staining of IL-10, IL-4, IFN-γ and IL-17 in Tregs alone (gray columns) or after treatment with rhMGL-Fc (black columns). The results correspond to the mean obtained from ten independent experiments ± SD. (C) Flow cytometry analysis of Tregs cultured with or without rhMGL-Fc and soluble anti-CD3. Quantification of cell death was performed using annexin-V-FITC and propidium iodide-PE. The results correspond to the mean value ± SD of three donors. * Corresponds to p<0.05.
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pone.0132617.g003: MGL-CD45RA interaction reduces immunosuppressive cytokine production.(A) Real-time PCR analysis of T-bet, RORγT and GATA3 genes in Tregs alone (grey columns) or after treatment with rhMGL-Fc (black columns). Relative gene expression was normalized with B2M and RLP0 gene expression levels. The results correspond to the mean value ± SD of three donors. (B) Intracellular staining of IL-10, IL-4, IFN-γ and IL-17 in Tregs alone (gray columns) or after treatment with rhMGL-Fc (black columns). The results correspond to the mean obtained from ten independent experiments ± SD. (C) Flow cytometry analysis of Tregs cultured with or without rhMGL-Fc and soluble anti-CD3. Quantification of cell death was performed using annexin-V-FITC and propidium iodide-PE. The results correspond to the mean value ± SD of three donors. * Corresponds to p<0.05.

Mentions: We also investigated whether the reduced Treg suppression observed might be attributed to a conversion toward another specific phenotype (Th1, Th2, and Th17). cDNA isolated by Tregs alone and Tregs treated with rhMGL-Fc were analysed by RT-PCR for the expression of T-bet, GATA-3 and RORγt, which are the transcription factors responsible for Th1, Th2 and Th17 cell commitment, respectively. We observed no significant differences between Tregs alone and Tregs after CD45RA stimulation, suggesting that MGL triggering did not favour the switching of Treg cells toward a specific Th lineage (Fig 3A).


The Macrophage Galactose-Type C-Type Lectin (MGL) Modulates Regulatory T Cell Functions.

Zizzari IG, Martufi P, Battisti F, Rahimi H, Caponnetto S, Bellati F, Nuti M, Rughetti A, Napoletano C - PLoS ONE (2015)

MGL-CD45RA interaction reduces immunosuppressive cytokine production.(A) Real-time PCR analysis of T-bet, RORγT and GATA3 genes in Tregs alone (grey columns) or after treatment with rhMGL-Fc (black columns). Relative gene expression was normalized with B2M and RLP0 gene expression levels. The results correspond to the mean value ± SD of three donors. (B) Intracellular staining of IL-10, IL-4, IFN-γ and IL-17 in Tregs alone (gray columns) or after treatment with rhMGL-Fc (black columns). The results correspond to the mean obtained from ten independent experiments ± SD. (C) Flow cytometry analysis of Tregs cultured with or without rhMGL-Fc and soluble anti-CD3. Quantification of cell death was performed using annexin-V-FITC and propidium iodide-PE. The results correspond to the mean value ± SD of three donors. * Corresponds to p<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493043&req=5

pone.0132617.g003: MGL-CD45RA interaction reduces immunosuppressive cytokine production.(A) Real-time PCR analysis of T-bet, RORγT and GATA3 genes in Tregs alone (grey columns) or after treatment with rhMGL-Fc (black columns). Relative gene expression was normalized with B2M and RLP0 gene expression levels. The results correspond to the mean value ± SD of three donors. (B) Intracellular staining of IL-10, IL-4, IFN-γ and IL-17 in Tregs alone (gray columns) or after treatment with rhMGL-Fc (black columns). The results correspond to the mean obtained from ten independent experiments ± SD. (C) Flow cytometry analysis of Tregs cultured with or without rhMGL-Fc and soluble anti-CD3. Quantification of cell death was performed using annexin-V-FITC and propidium iodide-PE. The results correspond to the mean value ± SD of three donors. * Corresponds to p<0.05.
Mentions: We also investigated whether the reduced Treg suppression observed might be attributed to a conversion toward another specific phenotype (Th1, Th2, and Th17). cDNA isolated by Tregs alone and Tregs treated with rhMGL-Fc were analysed by RT-PCR for the expression of T-bet, GATA-3 and RORγt, which are the transcription factors responsible for Th1, Th2 and Th17 cell commitment, respectively. We observed no significant differences between Tregs alone and Tregs after CD45RA stimulation, suggesting that MGL triggering did not favour the switching of Treg cells toward a specific Th lineage (Fig 3A).

Bottom Line: In this study, we demonstrate that Treg immunosuppressive activity can be modulated by the cross-linking between the CD45RA expressed by Tregs and the C-type lectin MGL.This effect can be attributed to changes in CD45RA and TCR signalling through the inhibition of Lck and inactivation of Zap-70, an increase in the Foxp3 methylation status and, ultimately, the reduced production of suppressive cytokines.These results indicate a role of MGL as an immunomodulator within the tumour microenvironment interfering with Treg functions, suggesting its possible use in the design of anticancer vaccines.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medicine, "Sapienza" University, Rome, Italy.

ABSTRACT
Regulatory T cells (Tregs) are physiologically designed to prevent autoimmune disease and maintain self-tolerance. In tumour microenvironments, their presence is related to a poor prognosis, and they influence the therapeutic outcome due to their capacity to suppress the immune response by cell-cell contact and to release immunosuppressive cytokines. In this study, we demonstrate that Treg immunosuppressive activity can be modulated by the cross-linking between the CD45RA expressed by Tregs and the C-type lectin MGL. This specific interaction strongly decreases the immunosuppressive activity of Tregs, restoring the proliferative capacity of co-cultured T lymphocytes. This effect can be attributed to changes in CD45RA and TCR signalling through the inhibition of Lck and inactivation of Zap-70, an increase in the Foxp3 methylation status and, ultimately, the reduced production of suppressive cytokines. These results indicate a role of MGL as an immunomodulator within the tumour microenvironment interfering with Treg functions, suggesting its possible use in the design of anticancer vaccines.

No MeSH data available.


Related in: MedlinePlus