Publicación:
Single-cell deep phenotyping of IgG-secreting cells for high-resolution immune monitoring
Single-cell deep phenotyping of IgG-secreting cells for high-resolution immune monitoring
| dc.contributor.author | Eyer K. | es_PE |
| dc.contributor.author | Doineau R.C.L. | es_PE |
| dc.contributor.author | Castrillon C.E. | es_PE |
| dc.contributor.author | Briseño-Roa L. | es_PE |
| dc.contributor.author | Menrath V. | es_PE |
| dc.contributor.author | Mottet G. | es_PE |
| dc.contributor.author | England P. | es_PE |
| dc.contributor.author | Godina A. | es_PE |
| dc.contributor.author | Brient-Litzler E. | es_PE |
| dc.contributor.author | Nizak C. | es_PE |
| dc.contributor.author | Jensen A. | es_PE |
| dc.contributor.author | Griffiths A.D. | es_PE |
| dc.contributor.author | Bibette J. | es_PE |
| dc.contributor.author | Bruhns P. | es_PE |
| dc.contributor.author | Baudry J. | es_PE |
| dc.date.accessioned | 2024-05-30T23:13:38Z | |
| dc.date.available | 2024-05-30T23:13:38Z | |
| dc.date.issued | 2017 | |
| dc.description | We would like to thank the Institut Pierre-Gilles de Gennes (IPGG) for use of clean room facilities and the laser engraver (CII08, Axyslaser), and Pfizer for the generous gift of TT-Alexa488, and CHO cell lines secreting TT4, TT7, and TT10. TT11 and TT27 antibodies are Pfizer proprietary antibodies isolated from their collaboration with HiFiBiO. We thank Pfizer (M. Holsti, G. Cheung, and W. Somers) as well as HiFiBiO Team (A. Gérard, A. Woolfe, M. Reichen, A. Poitou, S. Essonno, R. Kumar, S. Ellouze, K. Grosselin, B. Shen, and C. Brenan) for identification, rapid cloning, and validation of TT11 and TT27 antibodies. This work received support from the French Investissements d'Avenir program under the grant agreements ANR-10-NANO-02, ANR-10-IDEX-0001-02 PSL, ANR-10-LABX-31 and ANR-10- EQPX-34, by the French Agence Nationale de la Recherche (ANR-14-CE16-0011 project DROPmAbs), from Région Ile-de-France (DIM NanoK) and by the Institut Carnot Pasteur Maladies Infectieuses. K.E. acknowledges financial support from the 'Fondation Pierre-Gilles de Gennes', and the 'Swiss National Science Foundation' and the 'Society in Science—The Branco Weiss Fellowship'. C.C. acknowledges financial support from CONCYTEC, Peru. We would like to further acknowledge R. Henson for making the dscatter function for Matlab publicly available, M. Spitzer, J. Wildenhain, J. Rappsilber and M. Tyers for BoxPlotR that has been used to create the box plots, and B. Iannascoli (Unit of Antibodies in Therapy and Pathology, Institut Pasteur) for help with antibody production and cell lines. | |
| dc.description.abstract | Studies of the dynamics of the antibody-mediated immune response have been hampered by the absence of quantitative, high-throughput systems to analyze individual antibody-secreting cells1,2,3,4,5. Here we describe a simple microfluidic system, DropMap, in which single cells are compartmentalized in tens of thousands of 40-pL droplets and analyzed in two-dimensional droplet arrays using a fluorescence relocation-based immunoassay. Using DropMap, we characterized antibody-secreting cells in mice immunized with tetanus toxoid (TT) over a 7-week protocol, simultaneously analyzing the secretion rate and affinity of IgG from over 0.5 million individual cells enriched from spleen and bone marrow. Immunization resulted in dramatic increases in the range of both single-cell secretion rates and affinities, which spanned at maximum 3 and 4 logs, respectively. We observed differences over time in dynamics of secretion rate and affinity within and between anatomical compartments. This system will not only enable immune monitoring and optimization of immunization and vaccination protocols but also potentiate antibody screening6,7. | |
| dc.description.sponsorship | Consejo Nacional de Ciencia, Tecnología e Innovación Tecnológica - Concytec | |
| dc.identifier.doi | https://doi.org/10.1038/nbt.3964 | |
| dc.identifier.scopus | 2-s2.0-85031105486 | |
| dc.identifier.uri | https://hdl.handle.net/20.500.12390/631 | |
| dc.language.iso | eng | |
| dc.publisher | Nature Publishing Group | |
| dc.relation.ispartof | Nature Biotechnology | |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.subject | Physiology | |
| dc.subject | Antibodies | es_PE |
| dc.subject | Cytology | es_PE |
| dc.subject | Drops | es_PE |
| dc.subject | Immunization | es_PE |
| dc.subject | Droplet arrays | es_PE |
| dc.subject | High resolution | es_PE |
| dc.subject | High throughput systems | es_PE |
| dc.subject | Immune response | es_PE |
| dc.subject | Individual cells | es_PE |
| dc.subject | Micro fluidic system | es_PE |
| dc.subject.ocde | https://purl.org/pe-repo/ocde/ford#1.06.01 | |
| dc.title | Single-cell deep phenotyping of IgG-secreting cells for high-resolution immune monitoring | |
| dc.type | info:eu-repo/semantics/article | |
| dspace.entity.type | Publication | |
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