• Expert ANR

• Membre d'un pool d'experts
• Direction d'équipe
• Responsable de formations
• Responsable de diplôme (M2)

• Physique/Matière Condensée/Science des matériaux
  

Adaptive immune response is part of the dynamic changes that accompany motoneuron loss in amyotrophic lateral sclerosis (ALS). CD4+ T cells that regulate a protective immunity during the neu- rodegenerative process have received the most attention. CD8+ T cells are also observed in the spinal cord of patients and ALS mice although their contribution to the disease still remains elusive. Here, we found that activated CD8+ T lymphocytes infiltrate the central nervous system (CNS) of a mouse model of ALS at the symptomatic stage. Selective ablation of CD8+ T cells in mice expressing the ALS- associated superoxide dismutase-1 (SOD1)G93A mutant decreased spi- nal motoneuron loss. Using motoneuron-CD8+ T cell coculture sys- tems, we found that mutant SOD1-expressing CD8+ T lymphocytes selectively kill motoneurons. This cytotoxicity activity requires the recognition of the peptide-MHC-I complex (where MHC-I represents major histocompatibility complex class I). Measurement of interac- tion strength by atomic force microscopy-based single-cell force spec- troscopy demonstrated a specific MHC-I-dependent interaction between motoneuron and SOD1G93A CD8+ T cells. Activated mutant SOD1 CD8+ T cells produce interferon-γ, which elicits the expression of the MHC-I complex in motoneurons and exerts their cytotoxic function through Fas and granzyme pathways. In addition, analysis of the clonal diversity of CD8+ T cells in the periphery and CNS of ALS mice identified an antigen-restricted repertoire of their T cell receptor in the CNS. Our results suggest that self-directed immune response takes place during the course of the disease, contributing to the selective elimination of a subset of motoneurons in ALS.

Quantum sensing with diamond

Microgels display specific features attracting a high research interest in several applications such as bioactives, drug delivery or protein purification. Whey protein microgels (WPM) are obtained by promoting whey protein intramolecular cross-linking through heat treatment. Temperature-induced protein denaturation and aggregation involves several kinds of interactions such as electrostatic repulsions and disulfide bonds. In the present work, the WPM particles topography and nanomechanical properties were investigated at native pH (6.5) and acid pH (5.5 and 3.0) by atomic force microscopy (AFM) in contact mode in a liquid environment. Prior to AFM analysis, WPM particles were captured on a gold substrate via low energy interactions by means of specific monoclonal antibodies. The 2D AFM images clearly showed a swelling of WPM particles induced by pH decrease. At native pH, they displayed an average width and height of 192 ± 27 nm and 44 ± 9 nm, respectively. A decrease in pH to 5.5 led to a significant (p<0.05) increase in WPM particles width (282 ± 9 nm) and height (101 ± 1 nm). At pH 3.0, a further increase in WPM size (a width of 420 ± 25 nm and a height of 78 ± 1 nm). The AFM elasticity (E) data showed a significant (p<0.05) increase in stiffness at pH 5.5 (E: 199 ± 9 kPa) and pH 3.0 (E: 187 ± 12 kPa) compared to native pH (E: 12 ± 1 kPa). These findings indicate that the mechanical profiles of WPM network directly varied with pH decrease. The WPM topographic and nanomechanical changes induced by acidification were most likely due to substantial changes in the shape and the structure of WPM particles. These strengthened internally crosslinked structures, modified by acidification, could display interesting encapsulation properties, providing an additional proof for their use as nanovectors monitoring bioactives release to the desired target after ingestion.

Atomic force microscopy-based nanomechanical methods were directly applied in order to elucidate important questions related to brain metastasis formation and amyotrophic lateral sclerosis (ALS). Force mapping with single cell force spectroscopy was combined in order to gain direct insight into the surface “screening” process of tumor cells during their extravasation into the brain parenchyma. Furthermore, comparative SCFS measurements were performed between three melanoma cell types (WM35, A2058 and A375), showing altered invasive characteristics, and blood vessel lining endothelials in order to examine how the metastatic potential relates to tumor cell’s autonomous and inter-cellular nanomechanical properties. In case of ALS by comparing the adhesion of CD8+ cytotoxic T cells isolated from wild-type as well as SOD1G93A mutant mice against wild-type motor neurons we found that cytotoxic T cells are active contributors of neurodegeneration in the development of ALS. In addition, nanomechanical mapping of wild-type and SOD1G93A mutant mice derived single elongated myoblasts and multinuclear myotubes revealed differences in elasticity during early differentiation stage into myotubes. These findings provide valuable knowledge to the understanding of brain metastasis formation and ALS pathogenesis.