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1000 Titel
  • Unveiling Interindividual Variability of Human Fibroblast Innate Immune Response Using Robust Cell-Based Protocols
1000 Autor/in
  1. Chansard, Audrey |
  2. Dubrulle, Nelly |
  3. Poujol de Molliens, Mathilde |
  4. Falanga, Pierre B. |
  5. Stephen, Tharshana |
  6. Hasan, Milena |
  7. van Zandbergen, Ger |
  8. Aulner, Nathalie |
  9. Shorte, Spencer |
  10. David-Watine, Brigitte |
1000 Mitwirkende/r
  1. Milieu Intérieur Consortium |
1000 Erscheinungsjahr 2021
1000 Publikationstyp
  1. Artikel |
1000 Online veröffentlicht
  • 2021-01-11
1000 Erschienen in
1000 Quellenangabe
  • 11:569331
1000 FRL-Sammlung
1000 Copyrightjahr
  • 2021
1000 Lizenz
1000 Verlagsversion
  • https://doi.org/10.3389/fimmu.2020.569331 |
  • https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7829859/ |
1000 Ergänzendes Material
  • https://www.frontiersin.org/articles/10.3389/fimmu.2020.569331/full#h14 |
1000 Publikationsstatus
1000 Begutachtungsstatus
1000 Sprache der Publikation
1000 Abstract/Summary
  • The LabEx Milieu Interieur (MI) project is a clinical study centered on the detailed characterization of the baseline and induced immune responses in blood samples from 1,000 healthy donors. Analyses of these samples has lay ground for seminal studies on the genetic and environmental determinants of immunologic variance in a healthy cohort population. In the current study we developed in vitro methods enabling standardized quantification of MI-cohort-derived primary fibroblasts responses. Our results show that in vitro human donor cohort fibroblast responses to stimulation by different MAMPs analogs allows to characterize individual donor immune-phenotype variability. The results provide proof-of-concept foundation to a new experimental framework for such studies. A bio-bank of primary fibroblast lines was generated from 323 out of 1,000 healthy individuals selected from the MI-study cohort. To study inter-donor variability of innate immune response in primary human dermal fibroblasts we chose to measure the TLR3 and TLR4 response pathways, both receptors being expressed and previously studied in fibroblasts. We established high-throughput automation compatible methods for standardized primary fibroblast cell activation, using purified MAMPS analogs, poly I:C and LPS that stimulate TLR3 and TLR4 pathways respectively. These results were in turn compared with a stimulation method using infection by HSV-1 virus. Our “Add-only” protocol minimizes high-throughput automation system variability facilitating whole process automation from cell plating through stimulation to recovery of cell supernatants, and fluorescent labeling. Images were acquired automatically by high-throughput acquisition on an automated high-content imaging microscope. Under these methodological conditions standardized image acquisition provided for quantification of cellular responses allowing biological variability to be measured with low system noise and high biological signal fidelity. Optimal for automated analysis of immuno-phenotype of primary human cell responses our method and experimental framework as reported here is highly compatible to high-throughput screening protocols like those necessary for chemo-genomic screening. In context of primary fibroblasts derived from donors enrolled to the MI-clinical-study our results open the way to assert the utility of studying immune-phenotype characteristics relevant to a human clinical cohort.
1000 Sacherschließung
lokal NF-κB
lokal TLR (Toll like receptors)
lokal FACS
lokal innate immunity
lokal human primary cells
lokal HSV-1
lokal cytokines
gnd 4739713-5 Angeborene Immunität
lokal immortalization
1000 Fächerklassifikation (DDC)
1000 Liste der Beteiligten
  1. https://frl.publisso.de/adhoc/uri/Q2hhbnNhcmQsIEF1ZHJleSA=|https://orcid.org/0000-0003-0437-925X|https://frl.publisso.de/adhoc/uri/UG91am9sIGRlIE1vbGxpZW5zLCBNYXRoaWxkZSAg|https://frl.publisso.de/adhoc/uri/RmFsYW5nYSwgUGllcnJlIEIuIA==|https://orcid.org/0000-0002-6560-656X|https://orcid.org/0000-0002-0879-7099|https://orcid.org/0000-0001-5218-4962|https://orcid.org/0000-0002-5800-1853|https://orcid.org/0000-0002-2125-8663|https://orcid.org/0000-0002-3292-0042|https://frl.publisso.de/adhoc/uri/TWlsaWV1IEludMOpcmlldXIgQ29uc29ydGl1bSA=
1000 Label
1000 Förderer
  1. Agence Nationale de la Recherche |
  2. Agence Nationale de la Recherche |
  3. Agence Régionale de Santé Île-de-France |
  4. Infrastructures en Biologie Santé et Agronomie |
  5. Institut Pasteur |
1000 Fördernummer
  1. ANR-10-LABX-62-IBEID; ANR 10-LBX-73-REVIVE; ANR-10-LABX-69-01
  2. ANR-10-INSB-04-01
  3. -
  4. -
  5. -
1000 Förderprogramm
  1. Investissements d’Avenir programme; Laboratoire Revive; Laboratoire d’Excellence Milieu interieur
  2. France BioImaging
  3. Domaine d’interêt majeur One Health, DIM1Health
  4. -
  5. -
1000 Dateien
1000 Förderung
  1. 1000 joinedFunding-child
    1000 Förderer Agence Nationale de la Recherche |
    1000 Förderprogramm Investissements d’Avenir programme; Laboratoire Revive; Laboratoire d’Excellence Milieu interieur
    1000 Fördernummer ANR-10-LABX-62-IBEID; ANR 10-LBX-73-REVIVE; ANR-10-LABX-69-01
  2. 1000 joinedFunding-child
    1000 Förderer Agence Nationale de la Recherche |
    1000 Förderprogramm France BioImaging
    1000 Fördernummer ANR-10-INSB-04-01
  3. 1000 joinedFunding-child
    1000 Förderer Agence Régionale de Santé Île-de-France |
    1000 Förderprogramm Domaine d’interêt majeur One Health, DIM1Health
    1000 Fördernummer -
  4. 1000 joinedFunding-child
    1000 Förderer Infrastructures en Biologie Santé et Agronomie |
    1000 Förderprogramm -
    1000 Fördernummer -
  5. 1000 joinedFunding-child
    1000 Förderer Institut Pasteur |
    1000 Förderprogramm -
    1000 Fördernummer -
1000 Objektart article
1000 Beschrieben durch
1000 @id frl:6432411.rdf
1000 Erstellt am 2022-03-22T10:48:39.030+0100
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1000 Vgl. frl:6432411
1000 Oai Id
  1. oai:frl.publisso.de:frl:6432411 |
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