Contributions
Abstract: 237
Type: Free Communications
Abstract Category: Pathology and pathogenesis of MS - Immunology
Introduction: Numerous researches highlight the involvement of the CD8+T cells in Multiple Sclerosis (MS). Indeed, we have demonstrated that overrepresented CD8+T cells found at the lesion sites are also found in the Cerebro-Spinal Fluid (CSF) and to a lesser extent in the blood of MS patients, suggesting that CSF and blood may be a source of T cells involved in the disease process. However, to date we have not identified yet in the periphery the culprit CD8+T cells driving autoimmune inflammation nor their phenotype and/or function.
Aims and objectives: Our working hypothesis is that the cells able to provoke damages in the Central Nervous System (CNS), may have a specific phenotypic or functional pattern. To identify a specific cell subtype involved in MS pathology, we analyze the single CD8+T cells molecular signatures.
Methods: We isolate single memory CD8+T cells from the blood and the CSF from MS and Clinically Isolated Syndrome (CIS) patients and from the blood of Healthy Controls (HC) and patients with other inflammatory neurological diseases (including Neuromyelitis Optica Spectrum Disorders [NMOSD] and chronic meningitis). We then perform on each single-cells, qPCR of 96 well-chosen genes involved in the CD8+T cell biology. Finally, we define the transcriptional signature of each single-cell, we thus are able to define subsets of CD8+T cells specific of MS by a set of genes including CCR7, CD62L, CD58, CD94, transcription factors (TBET and TCF7) and cytotoxic molecules (Granzyme A/B, perforin and granulysin).
Results: We observe a clear clustering of single-cells from MS and CIS patients highly different from HC and patients with NMOSD or chronic meningitis, with a set of genes specifically over-expressed in unbiased analyses. We also find that, in MS patients, the CSF CD8+T cells and those from the blood have a highly similar profile with a high expression of migration markers allowing blood CD8+T cells to migrate to the CNS. Using T-SNE analysis, we identify a CD8+ effector memory T cell subtype with highly cytotoxic properties present only in MS patients.
Conclusions: Our data are the first to describe a specific molecular pattern of memory CD8+T cells from blood and CSF in MS patients. We are able to identify a specific cell subtype present only in MS patients made up of single cells strongly orienting toward an activated, effector and highly cytotoxic profile and expressing markers allowing them to migrate to the CNS.
Disclosure: E. Dugast received a grant from the ARSEP foundation
I. Vogel: Nothing to disclose
A. Garcia: Nothing to disclose
B. Nicol: Nothing to disclose
J. Morille: Nothing to disclose
M. Jacq-Foucher: Nothing to disclose
N. Jousset: Nothing to disclose
F. Le Frère: Nothing to disclose
S. Wiertlewski received consultancy fees, speaker fees, honoraria and clinical research grants (non-personal) from Biogen-Idec, Genzyme, Novartis, Merck, Roche, Sanofi-Aventis and Teva
A. Nicot: Nothing to disclose
L. Michel received honoraria as consultant from Biogen, Teva, Merck, Novartis, Roche and Sanofi-genzyme
L. Berthelot: Nothing to disclose
P.-A. Gourraud has received honoraria as consultant from Biogen, he is a founder of Methodomcis.com (2008). His research group is supported by the ATIP-Avenir INSERM program and the Region Pays de Loire ConnecTalent, ARSEP Foundation (France), and the Nantes University Foundation
S. Nataf received honoraria as consultant from Sanofi-genzyme and BASF
K. Tarte: Nothing to disclose
D.-A. Laplaud received honoraria and consulting fees from Biogen, Sanofi-Genzyme, Bayer, Novartis, Teva, Merck-Serono, Roche and Medday. Research supports from Biogen, Novartis, Sanofi-Genzyme, Roche and Medday
Abstract: 237
Type: Free Communications
Abstract Category: Pathology and pathogenesis of MS - Immunology
Introduction: Numerous researches highlight the involvement of the CD8+T cells in Multiple Sclerosis (MS). Indeed, we have demonstrated that overrepresented CD8+T cells found at the lesion sites are also found in the Cerebro-Spinal Fluid (CSF) and to a lesser extent in the blood of MS patients, suggesting that CSF and blood may be a source of T cells involved in the disease process. However, to date we have not identified yet in the periphery the culprit CD8+T cells driving autoimmune inflammation nor their phenotype and/or function.
Aims and objectives: Our working hypothesis is that the cells able to provoke damages in the Central Nervous System (CNS), may have a specific phenotypic or functional pattern. To identify a specific cell subtype involved in MS pathology, we analyze the single CD8+T cells molecular signatures.
Methods: We isolate single memory CD8+T cells from the blood and the CSF from MS and Clinically Isolated Syndrome (CIS) patients and from the blood of Healthy Controls (HC) and patients with other inflammatory neurological diseases (including Neuromyelitis Optica Spectrum Disorders [NMOSD] and chronic meningitis). We then perform on each single-cells, qPCR of 96 well-chosen genes involved in the CD8+T cell biology. Finally, we define the transcriptional signature of each single-cell, we thus are able to define subsets of CD8+T cells specific of MS by a set of genes including CCR7, CD62L, CD58, CD94, transcription factors (TBET and TCF7) and cytotoxic molecules (Granzyme A/B, perforin and granulysin).
Results: We observe a clear clustering of single-cells from MS and CIS patients highly different from HC and patients with NMOSD or chronic meningitis, with a set of genes specifically over-expressed in unbiased analyses. We also find that, in MS patients, the CSF CD8+T cells and those from the blood have a highly similar profile with a high expression of migration markers allowing blood CD8+T cells to migrate to the CNS. Using T-SNE analysis, we identify a CD8+ effector memory T cell subtype with highly cytotoxic properties present only in MS patients.
Conclusions: Our data are the first to describe a specific molecular pattern of memory CD8+T cells from blood and CSF in MS patients. We are able to identify a specific cell subtype present only in MS patients made up of single cells strongly orienting toward an activated, effector and highly cytotoxic profile and expressing markers allowing them to migrate to the CNS.
Disclosure: E. Dugast received a grant from the ARSEP foundation
I. Vogel: Nothing to disclose
A. Garcia: Nothing to disclose
B. Nicol: Nothing to disclose
J. Morille: Nothing to disclose
M. Jacq-Foucher: Nothing to disclose
N. Jousset: Nothing to disclose
F. Le Frère: Nothing to disclose
S. Wiertlewski received consultancy fees, speaker fees, honoraria and clinical research grants (non-personal) from Biogen-Idec, Genzyme, Novartis, Merck, Roche, Sanofi-Aventis and Teva
A. Nicot: Nothing to disclose
L. Michel received honoraria as consultant from Biogen, Teva, Merck, Novartis, Roche and Sanofi-genzyme
L. Berthelot: Nothing to disclose
P.-A. Gourraud has received honoraria as consultant from Biogen, he is a founder of Methodomcis.com (2008). His research group is supported by the ATIP-Avenir INSERM program and the Region Pays de Loire ConnecTalent, ARSEP Foundation (France), and the Nantes University Foundation
S. Nataf received honoraria as consultant from Sanofi-genzyme and BASF
K. Tarte: Nothing to disclose
D.-A. Laplaud received honoraria and consulting fees from Biogen, Sanofi-Genzyme, Bayer, Novartis, Teva, Merck-Serono, Roche and Medday. Research supports from Biogen, Novartis, Sanofi-Genzyme, Roche and Medday