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CHO DC-SIGN-N80A cell line

Invented by Alessandra Cambi
Invented at Radboud UMC
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N-glycosylation of DC-SIGN contributes to a more resistant adhesion site. (A) Zoom in to the averaged FD-curves of CHO cells expressing DC-SIGN-WT, DC-SIGN-N80A, DC-SIGN-ΔCRD, and DC-SIGN-ΔRep of Fig. 5B. Dotted lines indicate semi-linear fits to the slope before of the retraction curve until maximum force is reached at Fmax (i.e., contact region) from averaged retraction FD-curves DC-SIGN-WT and DC-SIGN-N80A. (B) The elasticity for cells expressing the different DC-SIGN-constructs, CHO-DC-SIGN-WT treated by lactose or CytD; measured by fitting the linear slope presented in (A). (C) Averaged FD-curves of CHO-DC-SIGN-WT cells untreated, cultured for 24 h in 100 mM lactose, or treated with 10 µM of the actin polymerization inhibitor CytD compared to CHO-DC-SIGN-N80A cells (N ≥ 15 cells; N ≥ 80 FD-curves). (D) Cartoon depicting the interaction between C. albicans and DC-SIGN and the role played by the N-glycans in strengthening pathogen-binding. When DC-SIGN nanoclusters bear N-glycans (1), they can laterally interact with transmembrane proteoglycans such as CD44, through galectin-mediated cross-linking. Molecules such as CD44 are almost stably associated with cortical actin, thus providing a structural scaffold that might facilitate DC-SIGN-actin interactions. The absence of the N-glycans (such as in N80A or after lactose treatment) prevents DC-SIGN-galectin interaction (2), thus hampering the possibility to firmly link to the actin cytoskeleton. The N-glycan-galectin network at the outer leaflet of the plasma membrane and, therefore, adds an extra level of binding strengthening during DC-SIGN-pathogen interactions, which is distinct yet connected to the cortical actin polymerization. Taken from Te Riet J, Joosten B, Reinieren-Beeren I, Figdor CG, Cambi A. N-glycan mediated adhesion strengthening during pathogen-receptor binding revealed by cell-cell force spectroscopy. Sci Rep. 2017;7(1):6713. Published 2017 Jul 27. doi:10.1038/s41598-017-07220-w
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Catalogue Number 160717
Antigen/Gene or Protein Targets Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin lacking the ligand binding domain N80.
Parental Line CHO
Synonyms DC-SIGN N80A
Host Hamster
Tissue Ovary
Disease Keywords HIV-1, Ebola Virus, Hepatitis C virus
Relevance Dendritic Cell-Specific ICAM-3-Grabbing Non-Integrin (DC-SIGN; CD209) is a type II plasma membrane PRR abundantly expressed in antigen-presenting cells such as dendritic cells (DCs) and activated macrophages. As member of the C-type Lectin Receptor (CLR) family, DC-SIGN binds a plethora of pathogens, ranging from viruses like HIV-1, ebola virus, and hepatitis C virus, to larger pathogens like Mycobacterium tuberculosis and Candida albicans. This N80A mutant lacks the N-glycosylation ligand binding domain.
Production Details CHO cell lines stably expressing DC-SIGN-N80 were established by Lipofectamin 2000 transfection, and were cultured in Ham’s F-12 medium supplemented with 10% heat-inactivated FBS, 1% Antibiotic Antimycotic Solution , and 0.5 mg/ml of the aminoglycoside antibiotic G418.
Conditional No
Research Area Cell Signaling & Signal Transduction, Immunology
Recommended Growing Conditions Ham’s F-12 medium supplemented with 10% heat-inactivated FBS, 1% Antibiotic Antimycotic Solution , and 0.5 mg/ml of the aminoglycoside antibiotic G418.

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Te Riet et al. 2017. Sci Rep. 7(1):6713. PMID: 28751750.

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References: 1 entry

Te Riet et al. 2017. Sci Rep. 7(1):6713. PMID: 28751750.

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Inventor Information

Inventors

Alessandra Cambi

Alessandra Cambi

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Invented at

Radboud UMC
Radboud UMC

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