Single-cell deep phenotyping of IgG-secreting cells for high-resolution immune monitoring

Autor: Jean Baudry, Patrick England, Klaus Eyer, Clément Nizak, Alexei Godina, Pierre Bruhns, Carlos Castrillon, Jérôme Bibette, Raphael Clement Li-Ming Doineau, Guillaume Mottet, Vera Menrath, Andrew D. Griffiths, Elodie Michele Christine Brient-Litzler, Luis Briseño-Roa, Allan Jensen
Přispěvatelé: Laboratoire Colloïdes et Matériaux Divisés (LCMD), Centre National de la Recherche Scientifique (CNRS)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL), Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), HiFiBio SAS, Centre de Recherche et Innovation Technologique (CITECH), Institut Pasteur [Paris], 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., ANR-10-IDEX-0001,PSL,Paris Sciences et Lettres(2010), ANR-10-EQPX-0034,IPGG,Institut Pierre Gilles de Gennes pour la microfluidique(2010), ANR-14-CE16-0011,DROP-mAbs,Analyse en profondeur de répertoires d'anticorps par microfluidique en gouttelettes couplé à du séquençage haut-débit pour le diagnostic et la découverte d'anticorps thérapeutiques(2014), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: Nature Biotechnology
Nature Biotechnology, Nature Publishing Group, 2017, 35 (10), pp.977-982. ⟨10.1038/nbt.3964⟩
Nature Biotechnology, 2017, 35 (10), pp.977-982. ⟨10.1038/nbt.3964⟩
ISSN: 1087-0156
DOI: 10.1038/nbt.3964⟩
Popis: International audience; 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 cells. 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 screening.
Databáze: OpenAIRE
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