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Anti cd1c clone l161

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Anti-CD1c (clone L161) is a monoclonal antibody that binds to the CD1c cell surface antigen. CD1c is a member of the CD1 family of antigen-presenting molecules that present lipid and glycolipid antigens to T cells.

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Lab products found in correlation

Lsrfortessa, by BD (3 mentions) Fixable viability dye, by Thermo Fisher Scientific (2 mentions) Permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Anti cd25 clone bc96, by BioLegend (2 mentions) Fix perm solution, by Thermo Fisher Scientific (2 mentions) Anti cd3 clone okt3 and clone ucht1 anti cd4 clone rpa t4 anti cd11c clone 3.9 anti cd14 clone m5e2 anti cd15 clone w6d3 anti cd16 clone 3g8 anti cd19 clone hib19 anti cd20 clone 2h7, by BioLegend (2 mentions) Anti cd45 clone hl30, by BioLegend (2 mentions) Anti cd45ra clone hi100 anti cd56 clone mem 188 anti cd86 clone it2.2 anti foxp3 clone 259d anti hla dr clone l243 anti sirpα clone se5a5, by BioLegend (2 mentions) Anti cd103 clone b ly7, by Thermo Fisher Scientific (2 mentions) Lsrfortessa flow cytometer, by BD (1 mentions) Cl097, by InvivoGen (1 mentions) Live dead yellow, by Thermo Fisher Scientific (1 mentions) Anti cd11b pe clone icrf44, by BD (1 mentions) Anti cd16 bv605 clone 3g8, by BD (1 mentions) Anti cd14 apc cy7 clone mφp 9, by BD (1 mentions) Lipopolysaccharides (lps), by Thermo Fisher Scientific (1 mentions) Anti cd83 clone hb15e, by BD (1 mentions) Anti cd86 clone it2, by BioLegend (1 mentions) Anti cd11b clone icrf44, by Thermo Fisher Scientific (1 mentions) Anti cd14 clone m5e2, by BioLegend (1 mentions)

5 protocols using anti cd1c clone l161

1

Retroviral Transduction of CD1c Mutants

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DNA constructs encoding single-residue CD1c mutants were synthesized (Life Technologies) and cloned into pMIG247 (link), and retroviruses were transduced into C1R cells. Surface expression of each mutant was confirmed by flow cytometry using anti-CD1c (clone L161, BioLegend), and cell lines were purified via cell sorting based on co-expression of eGFP and CD1c. C1R.CD1d cells were generated previously via similar means48 (link). C1R cells were co-cultured at a 1:1 ratio with J76.3C8 cells overnight, and CD69 expression on eGFP+ CD3+ cells was assessed using an LSRFortessa flow cytometer (BD Biosciences). CD69 median fluorescence intensity (MFI) is presented relative to activation levels induced by C1R.CD1c wild-type antigen-presenting cells.
Wun K.S., Reijneveld J.F., Cheng T.Y., Ladell K., Uldrich A.P., Le Nours J., Miners K.L., McLaren J.E., Grant E.J., Haigh O.L., Watkins T.S., Suliman S., Iwany S., Jimenez J., Calderon R., Tamara K.L., Leon S.R., Murray M.B., Mayfield J.A., Altman J.D., Purcell A.W., Miles J.J., Godfrey D.I., Gras S., Price D.A., Van Rhijn I., Moody D.B, & Rossjohn J. (2018). T cell autoreactivity directed toward CD1c itself rather than toward carried self lipids. Nature immunology, 19(4), 397-406.
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2

Cytokine profiling of stimulated PBMCs

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Cryo-recovered PBMC were cultured at 2.5×106 cells/mL in presence of 0.1 μg/mL LPS (eBioscience), 1 μg/mL of the synthetic toll-like receptor agonist CL097 (In vivogen), or no stimuli in round-bottom 96-well microplates at 37°C and 5% CO2 for five hours. Two patients with cirrhosis with insufficient cell numbers after cryo-recovery were not stimulated with LPS. After stimulation, supernatants were collected and cells were stained with a panel of antibodies comprising anti-CD1c (clone L161; BioLegend), anti-CD11b-PE (clone ICRF44; BD Biosciences), anti-CD16-BV605 (clone 3G8; BD Biosciences), anti-CD14-APC-Cy7 (clone MφP-9; BD Biosciences), anti-CD141-APC (clone 1A4; BD Biosciences), anti-CD33-PE-Cy7 (clone P67.6; BD Biosciences), anti-CD86-BV711 (clone 2331; BD Biosciences), anti-HLA-DR-BV786 (clone G46-6; BD Biosciences), anti-PD-L1-BUV395 (clone MIHI; BD Biosciences), anti-CD19-PerCP-Cy5.5 (clone SJ25C1; BD Biosciences), and Live/Dead™ Yellow (Life Technologies). Flow cytometric analysis was performed on a five-laser BD LSRFortessa (BD Biosciences). Gatings of cell populations were set based on an internal PBMC control that was run in each experiment.
Törnell A., Blick E., Al-Dury S., Grauers Wiktorin H., Waern J., Ringlander J., Einarsdottir S., Lindh M., Hellstrand K., Lagging M, & Martner A. (2023). Presence of MDSC associates with impaired antigen-specific T cell reactivity following COVID-19 vaccination in cirrhotic patients. Frontiers in Immunology, 14, 1287287.
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3

Comprehensive Immunophenotyping Protocol

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Cells were first stained with fixable viability dye (ThermoFisher Scientific, Paisley, UK), followed by staining with specific antibodies. Extracellular staining was performed in phosphate-buffered saline plus 0.1% bovine serum albumin and 0.05% sodium azide; intracellular staining was performed using fix/perm solution and permeabilization buffer (eBioscience, Hatfield, UK) as per the manufacturer’s protocol. Two percent mouse serum was added to cells to block nonspecific staining before addition of antibodies. The following antibodies were used in this study: anti-CD1c (clone L161); anti-CD3 (clone OKT3 and clone UCHT1); anti-CD4 (clone RPA-T4); anti-CD11c (clone 3.9); anti-CD14 (clone M5E2); anti-CD15 (clone W6D3); anti-CD16 (clone 3G8); anti-CD19 (clone HIB19); anti-CD20 (clone 2H7); anti-CD25 (clone BC96); anti-CD45 (clone Hl30); anti-CD45RA (clone HI100); anti-CD56 (clone MEM-188); anti-CD86 (clone IT2.2); anti-FOXP3 (clone 259D); anti-HLA-DR (clone L243); anti-SIRPα (clone SE5A5) (all from Biolegend, London, UK); anti-CD141 (clone 1A4, BD and clone M80, R&D Systems, Minneapolis, MN, USA); and anti-CD103 (clone B-Ly7, eBioscience). Production of the anti-integrin β8 (clone ADWA16) is described below.
Cells were analyzed using an LSR Fortessa or LSRII (BD, Oxford, UK), and data were analyzed using Flowjo software (Flowjo, OR, Ashland).
Fenton T., Kelly A., Shuttleworth E., Smedley C., Atakilit A., Powrie F., Campbell S., Nishimura S., Sheppard D., Levison S., Worthington J., Lehtinen M, & Travis M. (2016). Inflammatory cues enhance TGFβ activation by distinct subsets of human intestinal dendritic cells via integrin αvβ8. Mucosal immunology, 10(3), 624-634.
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4

Comprehensive Immunophenotyping Protocol

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Cells were first stained with fixable viability dye (ThermoFisher Scientific, Paisley, UK), followed by staining with specific antibodies. Extracellular staining was performed in phosphate-buffered saline plus 0.1% bovine serum albumin and 0.05% sodium azide; intracellular staining was performed using fix/perm solution and permeabilization buffer (eBioscience, Hatfield, UK) as per the manufacturer’s protocol. Two percent mouse serum was added to cells to block nonspecific staining before addition of antibodies. The following antibodies were used in this study: anti-CD1c (clone L161); anti-CD3 (clone OKT3 and clone UCHT1); anti-CD4 (clone RPA-T4); anti-CD11c (clone 3.9); anti-CD14 (clone M5E2); anti-CD15 (clone W6D3); anti-CD16 (clone 3G8); anti-CD19 (clone HIB19); anti-CD20 (clone 2H7); anti-CD25 (clone BC96); anti-CD45 (clone Hl30); anti-CD45RA (clone HI100); anti-CD56 (clone MEM-188); anti-CD86 (clone IT2.2); anti-FOXP3 (clone 259D); anti-HLA-DR (clone L243); anti-SIRPα (clone SE5A5) (all from Biolegend, London, UK); anti-CD141 (clone 1A4, BD and clone M80, R&D Systems, Minneapolis, MN, USA); and anti-CD103 (clone B-Ly7, eBioscience). Production of the anti-integrin β8 (clone ADWA16) is described below.
Cells were analyzed using an LSR Fortessa or LSRII (BD, Oxford, UK), and data were analyzed using Flowjo software (Flowjo, OR, Ashland).
Fenton T., Kelly A., Shuttleworth E., Smedley C., Atakilit A., Powrie F., Campbell S., Nishimura S., Sheppard D., Levison S., Worthington J., Lehtinen M, & Travis M. (2016). Inflammatory cues enhance TGFβ activation by distinct subsets of human intestinal dendritic cells via integrin αvβ8. Mucosal immunology, 10(3), 624-634.
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5

Comprehensive Multicolor Flow Cytometry Analysis

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Cells were analyzed by flow cytometry using the following anti-human antibodies: anti-CD14 (clone M5E2; BioLegend, London, UK), anti-CD86 (clone IT2.2; BioLegend), anti-CD83 (clone HB15e, BD Bioscience, Oxford, UK), anti-CD11c (clone 3.9; BioLegend), anti-CD1c (clone L161; BioLegend), anti-HLA-DR (clone, L243; BioLegend), anti-CD13 (clone WM15; BioLegend), anti-CD33 (clone P67.6; BioLegend), anti-CD141 (clone 1A4; BD Bioscience, Oxford, UK), and anti-CD11b (clone ICRF44; eBioscience). Dead cells were identified using fixable Viability dye Zombie UV (BioLegend, UK). The data were acquired on a LSRII flow cytometer or Fortessa (BD Bioscience) and analyzed using FACSDiva (BD Bioscience) or FlowJo software (Tree Star, Ashland, Oregon).
Melo-Gonzalez F., Fenton T.M., Forss C., Smedley C., Goenka A., MacDonald A.S., Thornton D.J, & Travis M.A. (2018). Intestinal mucin activates human dendritic cells and IL-8 production in a glycan-specific manner. The Journal of Biological Chemistry, 293(22), 8543-8553.
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