Phenotyping of Cultured Primary Immature Dendritic Cells

The phenotype of the cultured primary imDCs was confirmed at day 6 using flow cytometry analysis, essentially as described before [27] (link), [29] (link). The cells were analyzed using the primary labeled antibodies Lineage1-FITC, CD11c-APC, HLR-DR-V500, CD80-PE, CD83-PECy7 and CD86-V50 (Becton Dickinson). DC-SIGN levels of imDCs, Raji wild type, and Raji DC-SIGN cells were determined using an anti-DC-SIGN antibody and a secondary PE-labeled antibody (both R&D systems, MN, USA). Flow cytometry analysis was performed on a LSR-II (Becton Dickinson). Data was analyzed using Kaluza 1.2.

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Richter M.K., da Silva Voorham J.M., Torres Pedraza S., Hoornweg T.E., van de Pol D.P., Rodenhuis-Zybert I.A., Wilschut J, & Smit J.M. (2014). Immature Dengue Virus Is Infectious in Human Immature Dendritic Cells via Interaction with the Receptor Molecule DC-SIGN. PLoS ONE, 9(6), e98785.

Publication 2014

CD11c-APC CD80-PE anti-DC-SIGN antibody

Anti antibody Antibodies Antibody Cells Dc sign Fitc Flow cytometry Phenotype

Corresponding Organization :

Other organizations : University Medical Center Groningen, University of Groningen, Universidad de Antioquia

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Variable analysis

independent variables

Cell type (imDCs, Raji wild type, Raji DC-SIGN cells)

dependent variables

Phenotype of cultured primary imDCs confirmed at day 6 using flow cytometry analysis
DC-SIGN levels of imDCs, Raji wild type, and Raji DC-SIGN cells

control variables

Primary labeled antibodies (Lineage1-FITC, CD11c-APC, HLR-DR-V500, CD80-PE, CD83-PECy7, CD86-V50)
Anti-DC-SIGN antibody and secondary PE-labeled antibody
Flow cytometry analysis performed on a LSR-II (Becton Dickinson)
Data analyzed using Kaluza 1.2

controls

Positive control: Raji DC-SIGN cells
Negative control: Raji wild type cells

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