Composition

Involvement of 1 professor,1 associate professor and 2 engineers

Pr. Sophie Candon

Dr. Sahil Adriouch

Dr. Catalina Abad

M. Gaëtan Riou

  Facilities

INSTRUMENTS FOR ANALYSIS:
FACSCanto (BD Bioscience): 2-laser fully digital analyzer with up to 6-color simultaneous analysis,  equipped with a carrousel loader for automated analysis of 40 tubes.

CytoFlex(Beckman Coulter): 2-laser fully digital analyzer with up to 4-color simultaneous analysis, with capacity for microparticule detection (down to 200 nm).

LSRFortessa(BD Bioscience): This analyzer is a sophisticated analytical instrument, used for multiparametric analysis of cell samples. It is equipped with five lasers and is theoretically capable of up to 18-color analysis (in practice, 12-color simultaneous analysis).

INSTRUMENT FOR CELL SORTING:
FACSAria-IIIu(BD Bioscience): This fully digital high-speed cell sorter is used for sorting of cell samples based on multiple phenotypic parameters. Equipped with 4 lasers and capable of up to 12-color simultaneous analysis, it can also be used for single cell cloning in 96-well plates.

  Activity

Main activities

CyFlow offers the use of different multi-parametric flow cytometry analyzers and one high-speed FACS cell sorter.  The technical characteristics and advantages of each of these apparatus are described above. CyFlow equipment may be used for the following applications and studies:

• Cell immuno-phenotyping with the simultaneous measurement of up to 12 fluorescence parameters
• DNA content, ploidy and cell cycle analysis
• Assessment of apoptosis and cell death
• Cellular functional studies (calcium flux, oxidative burst, cell proliferation)
• Multiplexed and micro-particle-based analysis (fluorescence bead arrays)
• Detection of microparticule (down to 200 nm such as bacterias, yeasts, microalgae…)
• Molecularcytogenetics
• Micro-biological applications (detection of heterogeneous microbial population using nucleic acid probes, evaluation of sensitivity/resistance to drug)
• Cell sorting based on the simultaneous analysis of up to 12 fluorescence parameters at high speed (20,000 cells per second) and high purity (>95% purity)
• Single-cell flow cloning in 96-well plates for cell culture and molecular biology applications

The facility is also extending its activities to cell analysisusing other approaches than flow cytometry. It will soon provide a nanofluidic technology for single cell omics analysis.

Complementary activities

 The following activities can be provided on demand:

• Consulting in flow cytometry technologies and multiparametric cell sorting
• Training and education
• Scientific animations
• Industrial partnerships and support to start-up development
• Scientific information release and science promotion to public

  Main contributions

The CyFlow facility has allowed IRIB teams to obtain the following scientific results:

• Role of NAD in the functions, survival and phenotype of regulatory T cells
• Phenotypic, quantitative and functional characterization of regulatory T cells after treatment with biodrugs in human clinical trials
• Mechanism of immunological tolerance toward muscle-expressed antigens
• Phenotypic and functional characterization of muscle stem cells
• Phenotypic and functional characterization of cultured myoblast used in clinical trials
• Phenotypic, functional characterization and cell sorting of olfactory ensheathing cells from the olfactory bulb
• Phenotypic analysis and sorting of drosophila neurons
• Immune monitoring of T cell responses in secondary lymphoid organs and tissues following immunization in experimental models
• Analysis of B cell phenotype, number and function after treatment with biodrugs in pemphigus
• Analysis of B cell phenotype, number and function during hypersensitivity reaction to drugs (DRESS syndrome)

  Latest publications

2017

• Feuillette S, Delarue M, Riou G, Gaffuri AL, Wu J, Lenkei Z, Boyer O, Frébourg T, Campion D and Lecourtois M. Neuron-to-Neuron Transfer of FUS in Drosophila Primary Neuronal Culture Is Enhanced by ALS-Associated Mutations. J Mol Neurosci 2017, 62: 114-122.
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2016

• Belmonte L, Achamrah N, Nobis S, Guérin C, Riou G, Bôle-Feysot C, Boyer O, Richard V, Rego JC, Déchelotte P, Goichon A and Coëffier M. A role for intestinal TLR4-driven inflammatory response during activity-based anorexia. Sci Rep 2016, 6: 35813.
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• Hardet R, Chevalier B, Dupaty L, Naïmi Y, Riou G, Drouot L, Jean L, Salvetti A, Boyer O and Adriouch S. Oral-tolerization Prevents Immune Responses and Improves Transgene Persistence Following Gene Transfer Mediated by Adeno-associated Viral Vector. Mol Ther 2016, 24: 87-95.
  26265250

2014

• Bisson A, Fréret M, Drouot L, Jean L, Corre SL, Gourcerol G, Doucet C, Michot F, Boyer O and Lamacz M. Restoration of anal sphincter function after myoblast cell therapy in incontinent rats. Cell Transplant 2014
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2013

• Colliou N, Picard D, Caillot F, Calbo S, Le Corre S, Lim A, Lemercier B, Le Mauff B, Maho-Vaillant M, Jacquot S, Bedane C, Bernard P, Caux F, Prost C, Delaporte E, Doutre MS, Dreno B, Franck N, Ingen-Housz-Oro S, Chosidow O, Pauwels C, Picard C, Roujeau JC, Sigal M, Tancrede-Bohin E, Templier I, Eming R, Hertl M, D'Incan M, Joly P and Musette P. Long-term remissions of severe pemphigus after rituximab therapy are associated with prolonged failure of desmoglein B cell response. Sci Transl Med 2013, 5: 175ra30.
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• Mayeur A, Duclos C, Honoré A, Gauberti M, Drouot L, do Rego JC, Bon-Mardion N, Jean L, Vérin E, Emery E, Lemarchant S, Vivien D, Boyer O, Marie JP and Guérout N. Potential of olfactory ensheathing cells from different sources for spinal cord repair. PLoS One 2013, 8: e62860.
  23638158

2012

• Franck E, Bonneau C, Jean L, Henry JP, Lacoume Y, Salvetti A, Boyer O and Adriouch S. Immunological tolerance to muscle autoantigens involves peripheral deletion of autoreactive CD8+ T cells. PLoS One 2012, 7: e36444.
  22570714
• Liao H, Franck E, Fréret M, Adriouch S, Baba-Amer Y, Authier FJ, Boyer O and Gherardi RK. Myoinjury transiently activates muscle antigen-specific CD8+ T cells in lymph nodes in a mouse model. Arthritis Rheum 2012, 64: 3441-51.
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2011

• Adriouch S, Franck E, Drouot L, Bonneau C, Jolinon N, Salvetti A and Boyer O. Improved Immunological Tolerance Following Combination Therapy with CTLA-4/Ig and AAV-Mediated PD-L1/2 Muscle Gene Transfer. Front Microbiol 2011, 2: 199.
  22046170
• Guérout N, Duclos C, Drouot L, Abramovici O, Bon-Mardion N, Lacoume Y, Jean L, Boyer O and Marie JP. Transplantation of olfactory ensheathing cells promotes axonal regeneration and functional recovery of peripheral nerve lesion in rats. Muscle Nerve 2011, 43: 543-51.
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