Project description:Changes in Treg function are difficult to quantify due to the lack of Treg-exclusive markers in humans and the complexity of functional experiments. We sorted naive and memory human Tregs and conventional T cells, and identified genes that identify human Tregs regardless of their state of activation. We developed this Treg signature using Affymetrix human genome U133A 2.0 microarrays. To generate Tregs and Tconvs in multiple states of activation, naïve (CD4+CD25hiCD45RA+) and memory (CD4+CD25hiCD45RA-) Tregs, and naïve (CD4+CD25-CD45RA+) and memory (CD4+CD25-CD45RA-) Tconvs were sorted from blood of 7 healthy adults and RNA was isolated ex vivo or after stimulation for 40h, promoting activation-induced FOXP3 in Tconvs. The gene-expression profile of the eight cell subsets was analyzed.
Project description:Changes in Treg function are difficult to quantify due to the lack of Treg-exclusive markers in humans and the complexity of functional experiments. We sorted naive and memory human Tregs and conventional T cells, and identified genes that identify human Tregs regardless of their state of activation. We developed this Treg signature using Affymetrix human genome U133A 2.0 microarrays. To generate Tregs and Tconvs in multiple states of activation, naïve (CD4+CD25hiCD45RA+) and memory (CD4+CD25hiCD45RA-) Tregs, and naïve (CD4+CD25-CD45RA+) and memory (CD4+CD25-CD45RA-) Tconvs were sorted from blood of 7 healthy adults and RNA was isolated ex vivo or after stimulation for 40h, promoting activation-induced FOXP3 in Tconvs. The gene-expression profile of the eight cell subsets was analyzed. 7 adult healthy control samples were sorted into 4 subsets: naïve (CD4+CD25hiCD45RA+) and memory (CD4+CD25hiCD45RA-) Tregs, and naïve (CD4+CD25-CD45RA+) and memory (CD4+CD25-CD45RA-) Tconvs. These were used for RNA ex vivo and after 40h stimulation with anti-CD3/CD28 beads to induce an activation phenotype.
Project description:Disturbed expression of microRNAs (miRNAs) in regulatory T-cells (Tregs) leads to development of autoimmunity in experimental mouse models. However, the miRNA expression signature characterizing Tregs of autoimmune diseases, such as rheumatoid arthritis (RA) has not been determined yet. Moreover, the technical limitations prevented the analysis of such minute T-cell population as naive and memory Tregs. In this study we have used a microarray approach to comprehensively analyze miRNA expression signatures of naive Tregs (CD4+CD45RO-CD25++), memory Tregs (CD4+CD45RO+CD25+++), as well as conventional naive (CD4+CD45RO-CD25-) and memory (CD4+CD45RO+CD25-) T-cells (Tconvs) derived from peripheral blood of RA patients, and matched healthy controls. Differential expression of selected miRNAs was validated by TaqMan-based qRT-PCR. We found a positive correlation between increased expression of miR-451 in T-cells of RA patients and disease activity score (DAS28), ESR levels, and serum levels of IL-6. Moreover, we found characteristic, disease and treatment independent, global miRNA expression signatures defining naive Tregs, memory Tregs, naive Tconvs and memory Tconvs. The analysis allowed us to define miRNAs characteristic for a general naive phenotype (e.g. miR-92a), a general memory phenotype (e.g. miR-21, miR-155), and most importantly miRNAs specifically expressed in both naive and memory Tregs, defining as such the Treg phenotype (i.e. miR-146a, miR-3162, miR-1202, miR-1246a, and miR-4281). MicroRNA profiling was performed in four CD4+ T-cell subsets: naive Tconventional (CD3+CD8-CD45RO-CD25-), naive Tregulatory (CD3+CD8-CD45RO-CD25+), memory Tconventional (CD3+CD8-CD45RO+CD25-), and memory Tregulatory (CD3+CD8-CD45RO+CD25+) derived from 2 healthy controls, and 6 rheumatoid arthritis patients (total n=8).
Project description:Immune checkpoint inhibitors (ICIs) have transformed the treatment of many solid tumors. Still, their effectiveness in multiple myeloma (MM) remains underwhelming, highlighting the need to explore alternative approaches beyond conventional ICIs. Here, we identify CD161 as a novel inhibitory receptor on bone marrow resident memory CD8+ T cells (BM CD8+ TRMs), known for their sustained presence and vital role in local immune surveillance in MM BM tumor microenvironment. The CD161-CLEC2D axis, where CD161 interacts with CLEC2D on MM cells, mediates immune suppression and TRMs dysfunction. Blocking CD161 enhances TRMs function, including tissue residency, proliferation, and antitumor activity. Combining CD161 blockade with CAR-T therapy significantly alleviates CAR-T exhaustion, improving its therapeutic efficacy. Additionally, a similar CD161-driven exhaustion program is observed in other hematologic malignancies. These findings suggest that targeting the CD161-CLEC2D axis could offer a promising strategy to enhance MM treatment outcomes and CAR-T efficacy.
Project description:Immune checkpoint inhibitors (ICIs) have transformed the treatment of many solid tumors. Still, their effectiveness in multiple myeloma (MM) remains underwhelming, highlighting the need to explore alternative approaches beyond conventional ICIs. Here, we identify CD161 as a novel inhibitory receptor on bone marrow resident memory CD8+ T cells (BM CD8+ TRMs), known for their sustained presence and vital role in local immune surveillance in MM BM tumor microenvironment. The CD161-CLEC2D axis, where CD161 interacts with CLEC2D on MM cells, mediates immune suppression and TRMs dysfunction. Blocking CD161 enhances TRMs function, including tissue residency, proliferation, and antitumor activity. Combining CD161 blockade with CAR-T therapy significantly alleviates CAR-T exhaustion, improving its therapeutic efficacy. Additionally, a similar CD161-driven exhaustion program is observed in other hematologic malignancies. These findings suggest that targeting the CD161-CLEC2D axis could offer a promising strategy to enhance MM treatment outcomes and CAR-T efficacy.
Project description:Immune checkpoint inhibitors (ICIs) have transformed the treatment of many solid tumors. Still, their effectiveness in multiple myeloma (MM) remains underwhelming, highlighting the need to explore alternative approaches beyond conventional ICIs. Here, we identify CD161 as a novel inhibitory receptor on bone marrow resident memory CD8+ T cells (BM CD8+ TRMs), known for their sustained presence and vital role in local immune surveillance in MM BM tumor microenvironment. The CD161-CLEC2D axis, where CD161 interacts with CLEC2D on MM cells, mediates immune suppression and TRMs dysfunction. Blocking CD161 enhances TRMs function, including tissue residency, proliferation, and antitumor activity. Combining CD161 blockade with CAR-T therapy significantly alleviates CAR-T exhaustion, improving its therapeutic efficacy. Additionally, a similar CD161-driven exhaustion program is observed in other hematologic malignancies. These findings suggest that targeting the CD161-CLEC2D axis could offer a promising strategy to enhance MM treatment outcomes and CAR-T efficacy.