CD28 (L293) Monoclonal Antibodies Detecting Human Antigens

Transcription

1 Monoclonal Antibodies Detecting Human Antigens CD28 (L293) Form Catalog number Pure Pure PE PerCP-Cy Product availability varies by region. Contact BD Biosciences Customer Support or your local sales representative for information. RESEARCH APPLICATIONS DESCRIPTION Specificity Antigen distribution Research applications include studies of: Signal transduction pathways in T-lymphocyte activation 1-8 T-lymphocyte cytokine production 9,10-14 Cell-adhesion molecules linking B and T lymphocytes 4 T-lymphocyte functional subsets 3,4,10,15-20 Initiation and maintenance of chronic inflammatory lesions 3,21 Intrathymic T-lymphocyte development 8 Mixed lymphocyte responses stimulated by B7-1/B7-2 transfected cell lines 22 T-cell response in HIV infection 23 The CD28 24 antigen, a disulfide-linked homodimeric glycoprotein, with a molecular weight of 44 kilodaltons (kda), 1-5,15,25 is a cell-adhesion molecule (CAM) and functions as the ligand for CD80 (B7-1) and CD86 (B7-2) antigens, 26,27 which are present on activated B lymphocytes, 28 monocytes, 29 and dendritic cells. 30 Interaction of the CD28 antigen with CD80 or CD86 antigens, or both, co-stimulates CD2 and CD3 antigen/t-cell antigen receptor (TCR) dependent T-cell mediated proliferation and cytotoxicity. 31 The CD28 antigen is present on approximately 60% to 80% of peripheral blood T (CD3 + ) lymphocytes, 95% of CD4 + T lymphocytes, 50% of CD8 + T lymphocytes, and 5% of immature CD3 thymocytes. 3,4,16,17 T lymphocytes that lack the CD28 antigen reciprocally express the CD11b antigen. During thymocyte maturation, CD28 antigen expression increases from a low density on most CD4 + CD8 + immature thymocytes to a higher density on virtually all mature CD3 +, CD4 +, or CD8 + thymocytes. Cell activation further augments CD28 antigen density. Both CD4 + and CD8 + T-lymphocyte subsets can be separated into two functionally distinct subsets based on CD28 antigen density. These subsets differ in their cytotoxic and lymphokine-production capabilities. CD4 + lymphocytes that express high levels of the CD28 antigen lack CD3-mediated cytotoxic ability and produce only minimal amounts of cytokines, while CD4 + lymphocytes that express low levels of the CD28 antigen possess CD3-mediated cytotoxic ability and produce interleukin-2 (IL-2), γ-interferon (IFN-γ), and tumor necrosis factor α/β (TNF-α/β). 4,17 Expression of the CD28 antigen also divides the CD8 + lymphocyte subset into two functional groups. CD8 + CD28 + lymphocytes tend to have long-term proliferative capacity in vitro and in vivo, while CD8 + CD28 lymphocytes are thought to be effector cells with limited proliferative capacity. 32 For Research Use Only. Not for use in diagnostic or therapeutic procedures. Becton, Dickinson and Company BD Biosciences 2350 Qume Drive San Jose, CA USA bdbiosciences.com ResearchApplications@bd.com 12/

2 Functional characteristics Clone Composition Product configuration The CD28 antibody (clone L293), like other CD28 antibodies, is comitogenic for T lymphocytes in the presence of submitogenic concentrations of CD3 antibody, CD2 antibody, phytohemagglutinin (PHA), or phorbol esters, such as phorbol myristate acetate (PMA). 2-7 When used in combination with CD49d (clone L25), CD28 (clone L293) provides optimal co-stimulatory signals for detection of antigen-specific cytokine production. 33 The CD28 antibody (clone L293) has been shown to block the cognate interaction between T and B lymphocytes (both induction of B-lymphocyte differentiation and immunoglobulin production). 9 The CD28 antibody, clone L293, 22 is derived from hybridization of Sp2/0-Ag14 mouse myeloma cells with spleen cells from BALB/c mice immunized with the HPB-ALL T-cell line. The CD28 antibody is composed of mouse IgG 1 heavy chains and kappa light chains. The following reagents are supplied in phosphate buffered saline (PBS) containing a stabilizer and a preservative. Form Number of tests Volume per test (µl) a Amount provided (µg) Total volume (ml) Concentration (µg/ml) Stabilizer Preservative Pure ,000 N/A 0.01% Sodium azide Pure N/A 0.1% Sodium azide PE Gelatin 0.1% Sodium azide PerCP-Cy Gelatin 0.1% Sodium azide a. Volume required to stain 10 6 cells. PROCEDURE REPRESENTATIVE DATA Visit our website (bdbiosciences.com) or contact your local BD representative for the lyse/wash protocol for direct immunofluorescence. Flow cytometric analysis was performed on peripheral blood and gated on lymphocytes. Laser excitation was at 488 nm. Representative data analyzed with a BD FACS brand flow cytometer is shown in the following figure. PerCP-Cy5.5 PE HANDLING AND STORAGE WARNING Store vials at 2 C 8 C. Conjugated forms should not be frozen. Protect from exposure to light. Each reagent is stable until the expiration date shown on the bottle label when stored as directed. All biological specimens and materials coming in contact with them are considered biohazards. Handle as if capable of transmitting infection 34,35 and dispose of with proper precautions in accordance with federal, state, and local regulations. Never pipette by mouth. Wear suitable protective clothing, eyewear, and gloves Page 2

3 CHARACTERIZATION WARRANTY To ensure consistently high-quality reagents, each lot of antibody is tested for conformance with characteristics of a standard reagent. Representative flow cytometric data is included in this data sheet. Unless otherwise indicated in any applicable BD general conditions of sale for non-us customers, the following warranty applies to the purchase of these products. THE PRODUCTS SOLD HEREUNDER ARE WARRANTED ONLY TO CONFORM TO THE QUANTITY AND CONTENTS STATED ON THE LABEL OR IN THE PRODUCT LABELING AT THE TIME OF DELIVERY TO THE CUSTOMER. BD DISCLAIMS HEREBY ALL OTHER WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING WARRANTIES OF MERCHANTABILITY AND FITNESS FOR ANY PARTICULAR PURPOSE AND NONINFRINGEMENT. BD S SOLE LIABILITY IS LIMITED TO EITHER REPLACEMENT OF THE PRODUCTS OR REFUND OF THE PURCHASE PRICE. BD IS NOT LIABLE FOR PROPERTY DAMAGE OR ANY INCIDENTAL OR CONSEQUENTIAL DAMAGES, INCLUDING PERSONAL INJURY, OR ECONOMIC LOSS, CAUSED BY THE PRODUCT. REFERENCES 1. Reiter C. Cluster report: CD28. In: Knapp W, Dörken B, Gilks WR, et al, eds. Leucocyte Typing IV: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1989: van Lier RAW, Brouwer M, Aarden LA. Signals involved in T cell activation. T cell proliferation induced through the synergistic action of anti-cd28 and anti-cd2 monoclonal antibodies. Eur J Immunol. 1988;18(1): Hara R, Fu SM, Hansen JA. Human T-cell activation. II. A new activation pathway used by a major T-cell population via a disulfide-bonded dimer of a 44-kilodalton polypeptide (9.3 antigen). J Exp Med. 1985;161: June CH, Ledbetter JA, Linsley PS, Thompson CB. Role of the CD28 receptor in T-cell activation. Immunol Today. 1990;11: van Lier RAW, Brouwer M, De Jong R, Groot M, De Groot E, Aarden L. Functional properties of the human T-cell antigen CD28. In: Knapp W, Dörken B, Gilks W, et al, eds. Leucocyte Typing IV: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1989: Olive D, Cerdan C, Costello R, et al. CD28 and CTLA-4 cluster report. In: Schlossman SF, Boumsell L, Gilks W, et al, eds. Leucocyte Typing V: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1995: Gmünder H, Lesslauer W. A 45-kDa human T-cell membrane glycoprotein functions in the regulation of cell proliferative responses. Eur J Biochem. 1984;142: Turka LA, Linsley PS, Paine R, Schieven GL, Thompson CB, Ledbetter JA. Signal transduction via CD4, CD8, and CD28 in mature and immature thymocytes: implications for thymic selection. J Immunol. 1991;146: Waldrop SL, Davis KA, Maino VC, Picker LJ. Normal human CD4 + memory T cells display broad heterogeneity in their activation threshold for cytokine synthesis. J Immunol. 1998;161: Walker EB, Haley D, Miller W, et al. gp M peptide immunization of human lymphocyte antigen A2 + stage I-III melanoma patients induces significant increase in antigen-specific effector and long-term memory CD8 + T cells. Clin Cancer Res. 2004;10: Pitcher CJ, Quittner C, Peterson DM, et al. HIV-1 specific CD4 + T cells are detectable in most individuals with active HIV-1 infection, but decline with prolonged viral suppression. Nat Med. 1999;5: Maino VC, Picker LJ. Identification of functional subsets by flow cytometry: intracellular detection of cytokine expression. Cytometry. 1998;34: Suni MA, Picker LJ, Maino VC. Detection of antigen-specific T cell cytokine expression in whole blood by flow cytometry. J Immunol Methods. 1998;212: Maino VC. Rapid assessment of antigen induced cytokine expression in memory T cells by flow cytometry. Vet Immunol Immunopathol. 1998;63: Lum LG, Orcutt-Thordarson N, Seigneuret MC, Hansen JA. In vitro regulation of immunoglobulin synthesis by T-cell subpopulations defined by a new human T-cell antigen (9.3). Cell Immunol. 1982;72: Morishita Y, Sao H, Hansen JA, Martin PJ. A distinct subset of human CD4 + cells with a limited alloreactive T-cell receptor repertoire. J Immunol. 1989;143: Page

4 17. Rotteveel FTM, Kokkelink I, van Lier RA, et al. Clonal analysis of functionally distinct human CD4 + T cell subsets. J Exp Med. 1988;168: Topp MS, Riddell SR, Akatsuka Y, Jensen MC, Blattman JN, Greenberg PD. Restoration of CD28 expression in CD28 CD8 + memory effector T cells reconstitutes antigen-induced IL-2 production. J Exp Med. 2003;198: Werwitzke S, Tiede A, Drescher BE, Schmidt RE, Witte T. CD8β/CD28 expression defines functionally distinct populations of peripheral blood T lymphocytes. Clin Exp Immunol. 2003;133: Filaci G, Fravega M, Negrini S, et al. Nonantigen specific CD8 + T suppressor lymphocytes originate from CD8 + CD28 T cells and inhibit both T-cell proliferation and CTL function. Human Immunol. 2004;65: Linsley PS, Brady W, Grosmaire L, Aruffo A, Damle NK, Ledbetter JA. Binding of the B-cell activation antigen B7 to CD28 costimulates T-cell proliferation and interleukin-2 mrna accumulation. J Exp Med. 1991;173: Azuma M, Cayabyab M, Buck D, Phillips JH, Lanier LL. CD28 interaction with B7 costimulates primary allogeneic proliferative responses and cytotoxicity mediated by small, resting T lymphocytes. J Exp Med. 1992;175: Appay V, Papagno L, Spina CA, et al. Dynamics of T cell responses in HIV infection. J Immunol. 2002;168: McMichael AJ, Gotch FM. T-cell antigens: new and previously defined clusters. In: McMichael AJ, ed. Leucocyte Typing III: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1987: Hansen JA, Martin PJ, Nowinski RC. Monoclonal antibodies identifying a novel T-cell antigen and Ia antigens of human lymphocytes. Immunogenetics. 1980;10: Linsley PS, Green JL, Brady W, Bajorath J, Ledbetter JA, Peach R. Human B7-1 (CD80) and B7-2 (CD86) bind with similar avidities but distinct kinetics to CD28 and CTLA-4 receptors. Immunity. 1994;1: Levine BL, Ueda Y, Craighead N, Huang ML, June CH. CD28 ligands CD80 (B7-1) and CD86 (B7-2) induce long-term autocrine growth of CD4 + T cells and induce similar patterns of cytokine sevretion in vitro. Int Immunol. 1995;7: Freeman GJ, Freedman AS, Segil JM, Lee G, Whitman JF, Nadler LM. B7, a new member of the Ig superfamily with unique expression on activated and neoplastic B cells. J Immunol. 1989;143: Fleischer J, Soeth E, Reiling N, Grage-Griebenow E, Flad HD, Ernst M. Differential expression and function of CD80 (B7-1) and CD86 (B7-2) on human peripheral blood monocytes. Immunology. 1996;89: Caux C, Vanbervliet B, Massacrier C, et al. B70/B7-2 is identical to CD86 and is the major functional ligand for CD28 expressed on human dendritic cells. J Exp Med. 1994;180: Lanier LL, O'Fallon S, Somoza C, et al. CD80 (B7) and CD86 (B70) provide similar costimulatory signals for T cell proliferation, cytokine production, and generation of CTL. J Immunol. 1995;154: Azuma M, Phillips JH, Lanier LL. CD28 T lymphocytes: antigenic and functional properties. J Immunol. 1993;150: van Lier RA, Brouwer M, Zeijlemaker WR, Aarden L. Induction of T-cell proliferation by monoclonal antibodies: CD28 (Tp44) monoclonal antibodies link the CD3 and CD2 pathway of T-cell activation. In: McMichael AJ, ed. Leucocyte Typing III: White Cell Differentiation Antigens. New York, NY: Oxford University Press; 1986: Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline Third Edition. Wayne, PA: Clinical and Laboratory Standards Institute; CLSI document M29-A Centers for Disease Control. Perspectives in disease prevention and health promotion update: universal precautions for prevention of transmission of human immunodeficiency virus, hepatitis B virus, and other bloodborne pathogens in health-care settings. MMWR. 1988;37: PATENTS AND TRADEMARKS Cy is a trademark of GE Healthcare. This product is subject to proprietary rights of GE Healthcare and Carnegie Mellon University, and is made and sold under license from GE Healthcare. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and Page 4

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