Dc sign

Manufactured by Thermo Fisher Scientific

DC-SIGN is a type of lectin, a protein that binds to specific carbohydrates. It is primarily expressed on the surface of certain immune cells, such as dendritic cells. DC-SIGN plays a role in the recognition and binding of various pathogens, facilitating their interaction with host cells.

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Optimized Cell Lines for HIV Research

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The following reagent was obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH: C8166-45 (Cat#404) from Dr. Robert Gallo.^{29 (link)} HEK293T cells were obtained from ATCC (Manassas, VA). The DC-SIGN sequence expressed in C8166-DCSIGN, 293F-DCSIGN, and 293F-DCSIGN-CD4 cells is codon-optimized. FreeStyle 293-F (referred to herein as 293F) cells were obtained from Life Technologies (Grand Island, NY). All stable cell lines were subjected to limiting dilution cloning using a selectable marker; where applicable, clones were screened by surface staining for DC-SIGN, CXCR4, or CD4 (all antibodies from eBiosciences, San Diego, CA). The 293F-based cell lines were grown in Dulbecco’s modified Eagle medium (high glucose) supplemented with 10% fetal bovine serum, 2mM l-glutamine, and 1% nonessential amino acids. C8166-based cells were grown in CD Hybridoma Medium (Life Technologies) supplemented with 10% fetal bovine serum and 8mM l-glutamine.

Farley D.C., McCloskey L., Thorne B.A., Tareen S.U., Nicolai C.J., Campbell D.J., Bannister R., Stewart H.J., Pearson L.J., Moyer B.J., Robbins S.H., Zielinski L., Kim T., Radcliffe P.A., Mitrophanous K.A., Gombotz W.R., Miskin J.E, & Kelley-Clarke B. (2015). Development of a replication-competent lentivirus assay for dendritic cell-targeting lentiviral vectors. Molecular Therapy. Methods & Clinical Development, 2, 15017-.

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Multiparameter Flow Cytometry of Mouse Lung Leukocytes

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Isolated leukocytes from lungs of uninfected control as well as infected mice were stained with labeled monoclonal antibodies (mAbs) specific for cell surface markers or intracellular transcription factors or cytokines. mAbs for flow cytometry specific for CD11c (-Alexa-fluor 700), CD11b (-PECy7 or -APCCy7), MHCII (-PE), F4/80 (-PerCP-Cy5.5), DCsign (-APC), CD8(-APCCy7 -PECy5), CD4 (- FITC), Ly6c (-efluor-450), CD80 (-PE), CD86 (-PECy5), CD115 (-FITC), CD3 (-FITC) and CD3 (-efluor-450) were obtained from eBioscience and CD68 (-biotinylated plus streptavidin conjugated with Qdot605) was obteined from Invitrogen. The intracellular fixation and permeabilization buffer set of eBioscience was used to perform IFN-γ- APC, TNF-α-PE and NOS2-PE. To Tbet-PE, GATA-3-PE, RORγT-PE and Foxp3-PE transcription factors staining the Foxp3 staining buffer set was used (eBioscience).

Galvão-Filho B., de Castro J.T., Figueiredo M.M., Rosmaninho C.G., Antonelli L.R, & Gazzinelli R.T. (2019). The emergence of pathogenic TNF/iNOS producing dendritic cells (Tip-DCs) in a malaria model of acute respiratory distress syndrome (ARDS) is dependent on CCR4. Mucosal Immunology, 12(2), 312-322.

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