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Analysis of ICOS (CD278) Expression Left Panel - Two-color flow cytometric analysis of ICOS (CD278) expression on human peripheral blood lymphocytes. Whole blood was stained with FITC Mouse Anti-Human CD3 antibody (Cat. No. 561806/561807/555332) and either no antibody (BD Horizon™ R718 Autofluorescence Control; Top Plot) or BD Horizon™ R718 Hamster Anti-ICOS (CD278) antibody (Cat. No. 566990/566991; Bottom Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-color flow cytometric contour plots showing the correlated expression of ICOS (CD278) [or BD Horizon™ R718 Autofluorescence] versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Right Panel - Multicolor flow cytometric analysis of ICOS (CD278) expression on mouse thymocytes. Mouse thymocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553046/553047/561835) and PE Rat Anti-Mouse CD8a (Cat. No. 553033/553032/561095) antibodies, and either no antibody (BD Horizon™ R718 Autofluorescence Control; dashed line histograms) or BD Horizon™ R718 Hamster Anti-ICOS (CD278) antibody (solid line histograms) at 0.125 µg/test. The fluorescence histograms showing ICOS (CD278) expression (or BD Horizon™ R718 Autofluorescence) were derived from CD4 and CD8 gated events with the forward and side light-scatter characteristics of viable thymocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ R718 Armenian Hamster Anti-ICOS (CD278)
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Preparation And Storage
Recommended Assay Procedures
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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Companion Products
The C398.4A monoclonal antibody specifically binds to Inducible Costimulator (ICOS), which is also known as, CD278, Activation-inducible lymphocyte immunomediatory molecule (AILIM), or H4. ICOS is a type I transmembrane glycoprotein that forms a disulfide-linked homodimer and belongs to the CD28 family within the Ig superfamily. ICOS is expressed on either CD4+ or CD8+ single-positive mature thymocytes, T cells, or subsets of Innate Lymphoid Cells (ILC). Its expression is upregulated on activated T lymphocytes. ICOS is a costimulatory receptor that can bind to the B7-H2 ligand (CD275, ICOS-L) that is expressed on B cells, monocytes, macrophages, dendritic cells, and endothelial cells. ICOS plays a critical role in many types of T cell-dependent immunity. In the case of humoral immunity, for example, ICOS signaling is critical for the differentiation of T follicular helper (Tfh) cells and development of germinal centers. Although C398.4A was generated against mouse ICOS, this antibody reportedly crossreacts with human, rhesus, and rat ICOS.
The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively for BD Biosciences as a better alternative to Alexa Fluor® 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor® 700.
Development References (8)
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Araujo LM, Fert I, Jouhault Q, et al. Increased production of interleukin-17 over interleukin-10 by treg cells implicates inducible costimulator molecule in experimental spondyloarthritis. Arthritis Rheum. 2014; 66(9):2412-2422. (Clone-specific: Flow cytometry). View Reference
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Brenchley JM, Vinton C, Tabb B, et al. Differential infection patterns of CD4+ T cells and lymphoid tissue viral burden distinguish progressive and nonprogressive lentiviral infections.. Blood. 2012; 120(20):4172-81. (Clone-specific: Flow cytometry). View Reference
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Buonfiglio D, Bragardo M, Bonissoni S, et al. Characterization of a novel human surface molecule selectively expressed by mature thymocytes, activated T cells and subsets of T cell lymphomas. Eur J Immunol. 1999; 29(9):2863-2879. (Clone-specific: Flow cytometry, Immunohistochemistry, Immunoprecipitation, Radioimmunoassay). View Reference
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Buonfiglio D, Bragardo M, Redoglia V, et al. The T cell activation molecule H4 and the CD28-like molecule ICOS are identical. Eur J Immunol. 2000; 30(12):3463-3467. (Clone-specific: Blocking, Flow cytometry). View Reference
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Chen Y, Shen S, Gorentla BK, Gao J, Zhong XP. Murine regulatory T cells contain hyperproliferative and death-prone subsets with differential ICOS expression. J Immunol. 2012; 188(4):1698-1707. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Klatt NR, Vinton CL, Lynch RM et al. SIV infection of rhesus macaques results in dysfunctional T- and B-cell responses to neo and recall Leishmania major vaccination. Blood. 2011; 118(22):5803-5812. (Clone-specific: Flow cytometry). View Reference
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Redoglia V, Dianzani U, Rojo JM, et al. Characterization of H4: a mouse T lymphocyte activation molecule functionally associated with the CD3/T cell receptor. Eur J Immunol. 1996; 26(11):2781-2789. (Immunogen: Bioassay, (Co)-stimulation, Flow cytometry, Functional assay, Immunoprecipitation, Radioimmunoassay). View Reference
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Xu H, Wang X, Lackner AA, Veazey RS. PD-1(HIGH) Follicular CD4 T Helper Cell Subsets Residing in Lymph Node Germinal Centers Correlate with B Cell Maturation and IgG Production in Rhesus Macaques. Front Biosci. 2014; 5(85):1-7. (Clone-specific: Flow cytometry). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.