Limits...
Killing of myeloid APCs via HLA class I, CD2 and CD226 defines a novel mechanism of suppression by human Tr1 cells.

Magnani CF, Alberigo G, Bacchetta R, Serafini G, Andreani M, Roncarolo MG, Gregori S - Eur. J. Immunol. (2011)

Bottom Line: Notably, interaction between CD226 on Tr1 cells and their ligands on myeloid cells, leading to Tr1-cell activation, is necessary for defining Tr1-cell target specificity.We also showed that high frequency of GZB-expressing CD4(+) T cells is detected in tolerant patients and correlates with elevated occurrence of IL-10-producing CD4(+) T cells.In conclusion, the modulatory activities of Tr1 cells are not only due to suppressive cytokines but also to specific cell-to-cell interactions that lead to selective killing of myeloid cells and possibly bystander suppression.

View Article: PubMed Central - PubMed

Affiliation: San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, San Raffaele Scientific Institute, Milan, Italy.

Show MeSH

Related in: MedlinePlus

Tr1-mediated cytotoxicity requires CD18 adhesion and activation via CD2 and CD226. (A) Tr1-cell lines were co-cultured with freshly isolated allogeneic CD14+ and CD1c+ cells at 10:1 (E:T) ratio in the presence of anti-CD18 mAb or IgG1 isotype control, and degranulation in the presence of GZB was measured by co-expression of CD107a and GZB in CD4+ T cells. One donor representative of three donors tested in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells. (B) Tr1- and Th0-cell lines were co-cultured with p815 cells pre-incubated with anti-CD2 mAb at 10:1 (E:T) ratio, and degranulation was determined by co-expression of CD107a and GZB in CD4+ T cells. One donor representative of three donors performed in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells. (C) Tr1- and Th0-cell lines were co-cultured with p815 cells pre-incubated with anti-CD226 mAb at 10:1 (E:T) ratio, and degranulation was determined by co-expression of CD107a and GZB in CD4+ T cells. One donor out of five donors performed in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3116154&req=5

fig06: Tr1-mediated cytotoxicity requires CD18 adhesion and activation via CD2 and CD226. (A) Tr1-cell lines were co-cultured with freshly isolated allogeneic CD14+ and CD1c+ cells at 10:1 (E:T) ratio in the presence of anti-CD18 mAb or IgG1 isotype control, and degranulation in the presence of GZB was measured by co-expression of CD107a and GZB in CD4+ T cells. One donor representative of three donors tested in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells. (B) Tr1- and Th0-cell lines were co-cultured with p815 cells pre-incubated with anti-CD2 mAb at 10:1 (E:T) ratio, and degranulation was determined by co-expression of CD107a and GZB in CD4+ T cells. One donor representative of three donors performed in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells. (C) Tr1- and Th0-cell lines were co-cultured with p815 cells pre-incubated with anti-CD226 mAb at 10:1 (E:T) ratio, and degranulation was determined by co-expression of CD107a and GZB in CD4+ T cells. One donor out of five donors performed in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells.

Mentions: To dissect the molecular mechanism underlying the target specificity of Tr1 cells, we investigated adhesion and signaling molecules involved in Tr1-myeloid APC interaction. Both CD14+ and CD1c+ cells expressed higher levels of CD54 compared to T and B cells (not shown), and Tr1 cells expressed LFA-1 (CD18/CD11a), the CD54 ligand (Supporting Information Fig. 6). Addition of neutralizing anti-CD18 mAb blocked degranulation of Tr1-cell lines when co-cultured with CD14+ or CD1c+ cells (Fig. 6A). CD54 expression on target cells is specifically involved in the formation of a stable immunological synapse essential for cytotoxicity mediated by NK cells and CTLs 14. Therefore, our findings suggest that the high expression of CD54 on myeloid cells is responsible for the formation of a stable and prolong interaction with Tr1 cells, leading to lysis of the target cell.


Killing of myeloid APCs via HLA class I, CD2 and CD226 defines a novel mechanism of suppression by human Tr1 cells.

Magnani CF, Alberigo G, Bacchetta R, Serafini G, Andreani M, Roncarolo MG, Gregori S - Eur. J. Immunol. (2011)

Tr1-mediated cytotoxicity requires CD18 adhesion and activation via CD2 and CD226. (A) Tr1-cell lines were co-cultured with freshly isolated allogeneic CD14+ and CD1c+ cells at 10:1 (E:T) ratio in the presence of anti-CD18 mAb or IgG1 isotype control, and degranulation in the presence of GZB was measured by co-expression of CD107a and GZB in CD4+ T cells. One donor representative of three donors tested in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells. (B) Tr1- and Th0-cell lines were co-cultured with p815 cells pre-incubated with anti-CD2 mAb at 10:1 (E:T) ratio, and degranulation was determined by co-expression of CD107a and GZB in CD4+ T cells. One donor representative of three donors performed in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells. (C) Tr1- and Th0-cell lines were co-cultured with p815 cells pre-incubated with anti-CD226 mAb at 10:1 (E:T) ratio, and degranulation was determined by co-expression of CD107a and GZB in CD4+ T cells. One donor out of five donors performed in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3116154&req=5

fig06: Tr1-mediated cytotoxicity requires CD18 adhesion and activation via CD2 and CD226. (A) Tr1-cell lines were co-cultured with freshly isolated allogeneic CD14+ and CD1c+ cells at 10:1 (E:T) ratio in the presence of anti-CD18 mAb or IgG1 isotype control, and degranulation in the presence of GZB was measured by co-expression of CD107a and GZB in CD4+ T cells. One donor representative of three donors tested in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells. (B) Tr1- and Th0-cell lines were co-cultured with p815 cells pre-incubated with anti-CD2 mAb at 10:1 (E:T) ratio, and degranulation was determined by co-expression of CD107a and GZB in CD4+ T cells. One donor representative of three donors performed in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells. (C) Tr1- and Th0-cell lines were co-cultured with p815 cells pre-incubated with anti-CD226 mAb at 10:1 (E:T) ratio, and degranulation was determined by co-expression of CD107a and GZB in CD4+ T cells. One donor out of five donors performed in two independent experiments is shown. Numbers represent percentage of CD107a+GZB+ cells.
Mentions: To dissect the molecular mechanism underlying the target specificity of Tr1 cells, we investigated adhesion and signaling molecules involved in Tr1-myeloid APC interaction. Both CD14+ and CD1c+ cells expressed higher levels of CD54 compared to T and B cells (not shown), and Tr1 cells expressed LFA-1 (CD18/CD11a), the CD54 ligand (Supporting Information Fig. 6). Addition of neutralizing anti-CD18 mAb blocked degranulation of Tr1-cell lines when co-cultured with CD14+ or CD1c+ cells (Fig. 6A). CD54 expression on target cells is specifically involved in the formation of a stable immunological synapse essential for cytotoxicity mediated by NK cells and CTLs 14. Therefore, our findings suggest that the high expression of CD54 on myeloid cells is responsible for the formation of a stable and prolong interaction with Tr1 cells, leading to lysis of the target cell.

Bottom Line: Notably, interaction between CD226 on Tr1 cells and their ligands on myeloid cells, leading to Tr1-cell activation, is necessary for defining Tr1-cell target specificity.We also showed that high frequency of GZB-expressing CD4(+) T cells is detected in tolerant patients and correlates with elevated occurrence of IL-10-producing CD4(+) T cells.In conclusion, the modulatory activities of Tr1 cells are not only due to suppressive cytokines but also to specific cell-to-cell interactions that lead to selective killing of myeloid cells and possibly bystander suppression.

View Article: PubMed Central - PubMed

Affiliation: San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, San Raffaele Scientific Institute, Milan, Italy.

Show MeSH
Related in: MedlinePlus