Cd1c clone l161

Manufactured by BioLegend

CD1c (clone L161) is a monoclonal antibody that specifically binds to the CD1c antigen, which is a cell surface glycoprotein belonging to the CD1 family of antigen-presenting molecules. The core function of this antibody is to facilitate the detection and identification of CD1c-expressing cells in various research applications.

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Anti-CD1c (clone L161; (5 citations)

4 protocols using cd1c clone l161

Quantifying Costimulatory Molecule Expression on APCs

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Enriched APCs were stained with antibodies recognizing OX40L (clone ANC10G1; Ancell), ICOSL (clone 136726; R&D Bio-techne), or CD86 (clone FUN-1; R&D Bio-techne), or isotype control, and HLA-DR (clone LN3; eBiosciences), CD14 (clone 61D3; eBiosciences), CD11c (clone Bu15; BioLegend), CD1c (clone L161; BioLegend), CD304 (clone 12C2; BioLegend), and CD141 (clone AD5-14H12; Miltenyi Biotec). Samples were acquired on a FACS Verse instrument (BD Biosciences). Stain index was calculated to quantify the level of expression as follows: stain index = (MFI_sample − MFI_isotype)/(2 × SD_isotype), where MFI indicates mean fluorescence intensity, and SD indicates standard deviation.

Durand M., Walter T., Pirnay T., Naessens T., Gueguen P., Goudot C., Lameiras S., Chang Q., Talaei N., Ornatsky O., Vassilevskaia T., Baulande S., Amigorena S, & Segura E. (2019). Human lymphoid organ cDC2 and macrophages play complementary roles in T follicular helper responses. The Journal of Experimental Medicine, 216(7), 1561-1581.

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Phenotyping and Transcriptional Profiling of Immune Cells in Rheumatoid Arthritis

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Ex vivo and cultured PBMC were stained with APC-H7 (Tonbo Biosciences) or Brilliant Violet 405 (Molecular Probes) viability dye in the presence of different panels of monoclonal antibodies against lineage (Lin) (CD3 clone HIT3a, CD19 clone SJ25C1, CD20 clone 2H7, and CD56 clone 5.1H11), CD14 clone M5E2, CD16 clone 3G8, CD40 clone 5C3, CD86 clone IT2.2, ILT4 clone 42D1, HLA-DR clone L243, CD11c clone 3.9, CD1c clone L161, CD141 clone M80, CD64 clone 10.1, and PDL1 clone 29E.2A3 (BioLegend). Samples were analyzed on a Fortessa cytometer (BD Biosciences) at Centro Nacional de Investigaciones Cardiovasculares (CNIC, Madrid, Spain). Analysis of individual and multiparametric flow cytometry data was performed using FlowJo software (Tree Star Inc.).
For the transcriptional studies, viable human Lin^–CD14^–CD11c⁺HLADR⁺CD1c⁺ cDC, CD14^–CD11c⁺HLADR⁺CD141⁺ cDC, and total CD14⁺ Mo were sorted using a FACS Aria II sorter (BD Biosciences) from either PBMC from n = 4 untreated patients with RA and n = 4 HC, or SF from n = 3 individuals with RA and n = 3 suffering mechanic CPPD crystal–associated arthritis.

Delgado-Arévalo C., Calvet-Mirabent M., Triguero-Martínez A., Vázquez de Luis E., Benguría-Filippini A., Largo R., Calzada-Fraile D., Popova O., Sánchez-Cerrillo I., Tsukalov I., Moreno-Vellisca R., de la Fuente H., Herrero-Beaumont G., Ramiro A., Sánchez-Madrid F., Castañeda S., Dopazo A., González Álvaro I, & Martin-Gayo E. (2022). NLRC4-mediated activation of CD1c+ DC contributes to perpetuation of synovitis in rheumatoid arthritis. JCI Insight, 7(22), e152886.

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Flow Cytometry Analysis of PBMCs and CSF

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Flow cytometry analysis was performed using antihuman antibodies to BDCA-2 (clone 201A; BioLegend, San Diego, CA), CD33 (clone P67.6; BioLegend), CD14 (clone 61D3; eBioscience, San Diego, CA), Lyve-1 (clone 537028; R&D Systems, Minneapolis, MN), CD3 (clone SK7; BioLegend), CD19 (clone HIB19; eBioscience), CD16 (clone B73.1; BioLegend), CD1c (clone L161; BioLegend), Lox-1 (clone 15C4; BioLegend), and human leukocyte antigen DR (HLA-DR) (clone Immu-357; Beckman Coulter, Brea, CA). One million PBMCs and between 2 × 10⁴ and 40 × 10⁴ CSF cells were incubated with human-Fc block (BD Biosciences, San Jose, CA) for 10 minutes at RT and then with antibodies for 20 minutes at 4°C. Subsequently, the samples were washed with PBS + 2% FBS (flow buffer) for 5′, spun at 500g, and resuspended in 200 μL of flow buffer. The samples were run on a Gallios flow cytometer (Beckman Coulter Life Sciences) on the same day of the collection. The cells were analyzed using FlowJo software (Tree Star, Ashland, OR).

Esaulova E., Cantoni C., Shchukina I., Zaitsev K., Bucelli R.C., Wu G.F., Artyomov M.N., Cross A.H, & Edelson B.T. (2020). Single-cell RNA-seq analysis of human CSF microglia and myeloid cells in neuroinflammation. Neurology® Neuroimmunology & Neuroinflammation, 7(4), e732.

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Isolation and Characterization of Immune Cells

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Tonsil or lymph node samples were digested as described previously (Durand and Segura, 2016 (link)). In brief, DC subsets and macrophages were selected by density gradient, enriched by negative selection, and isolated by cell sorting on a FACS Aria instrument (BD Biosciences). Antibodies used were anti-CD11c (clone Bu15; BioLegend), HLA-DR (clone LN3; eBioscience), CD14 (clone 61D3; eBioscience), CD1c (clone L161; BioLegend), CD141 (clone AD5-14H12; Miltenyi Biotec), and CD123 (clone AC145; Miltenyi Biotec).

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