Human LAG-3 PE-conjugated Antibody Summary
Leu23-Leu450
Accession # P18627
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of LAG-3 in HEK293 Human Cell Line Transfected with Human LAG-3 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) human LAG-3 or (B) irrelevant transfectants and eGFP was stained with Goat Anti-Human LAG-3 Phycoerythrin-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB2319P). Quadrant markers were set based on control antibody staining (Catalog # IC108P, data not shown). View our protocol for Staining Membrane-associated Proteins.
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Detection of LAG-3 in Human PBMCs by Flow Cytometry. CD3+ PHA-treated peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD3e APC-conjugated Monoclonal Antibody (Catalog # FAB100A) and either (A) Goat Anti-Human LAG-3 PE-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB2319P) or (B) Normal Goat IgG Phycoerythrin Control (Catalog # IC108P). View our protocol for Staining Membrane-associated Proteins.
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Detection of Human LAG-3 by Flow Cytometry Human CD4+CD25−LAG3+ T cells suppressed antibody production. a Gating strategy of fluorescence-activated cell sorting analysis for CD4+CD25−LAG3+CD45RA− T cells (LAG3+ Tregs) and CD4+CD25+CD127dimCD45RA− T cells (CD25+ Tregs). b IL-10, IFN-gamma, and IL-17A protein levels in the culture supernatants after 3 days of incubation of the indicated T-cell subsets was determined by ELISA (n = 6). c B cells and TFH cells were cocultured with the indicated subsets for 12 days in the presence of SEB stimulation, and total IgG, IgM, and IgA production was determined by ELISA (n = 6). d B cells and TFH cells were cocultured with CD4+CD25−LAG3+CD49b− T cells for 12 days in the presence of SEB stimulation, and total IgG was determined by ELISA (n = 3). All error bars represent SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. All P values were calculated by one-way analysis of variance and Tukey’s multiple-comparisons test. CD Cluster of differentiation, ELISA Enzyme-linked immunosorbent assay, IFN-gamma Interferon-gamma, Ig Immunoglobulin, IL Interleukin, LAG3 Lymphocyte activation gene 3, RA Rheumatoid arthritis, SEB Staphylococcal enterotoxin B, TFH Follicular helper T cell, Treg Regulatory T cell Image collected and cropped by CiteAb from the following publication (https://arthritis-research.biomedcentral.com/articles/10.1186/s13075-017-1309-x), licensed under a CC-BY license. Not internally tested by R&D Systems.
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Detection of Human LAG-3 by Flow Cytometry Effect of abatacept on the CD4+CD25−LAG3+ T-cell subset in vitro. a Naive CD4+ T cells and antigen-presenting cells (APCs) taken from healthy donors were cultured with anti-CD3 antibody in the presence of abatacept, and FACS was conducted on day 4. Representative FACS data from three independent experiments are shown. Percentages of CD4+CD25−LAG3+ T cells in CD4+ T cells are presented in the bar graph (n = 3). b Quantitative real-time polymerase chain reaction analysis of EGR2, FAS, and CD274 (PDL1) mRNA expression in the cells that were induced from naive CD4+ T cells in the presence of abatacept (n = 3). c Sorted 7-amino actinomycin D-negative CD4+ T cells were cultured with B cells and TFH cells under staphylococcal enterotoxin B stimulation for 12 days, and total IgG production was determined by ELISA (n = 3). All error bars represent SD. ** P < 0.001; **** P < 0.0001. All P values were calculated by unpaired two-tailed Student’s t test. CD Cluster of differentiation 4, EGR2 Early growth response gene 2, FACS Fluorescence-activated cell sorting, GAPDH Glyceraldehyde 3-phosphate dehydrogenase, IgG Immunoglobulin G, LAG3 Lymphocyte activation gene 3, mRNA Messenger RNA, PDL1 Programmed death-ligand 1, TFH Follicular helper T cell Image collected and cropped by CiteAb from the following publication (https://arthritis-research.biomedcentral.com/articles/10.1186/s13075-017-1309-x), licensed under a CC-BY license. Not internally tested by R&D Systems.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, 2 to 8 °C as supplied.
Background: LAG-3
LAG-3 (Lymphocyte activation gene-3), also known as CD223, is a member of the immunoglobulin superfamily (IgSF). The mature LAG-3 protein is a 496 amino acid (aa) membrane protein with a 421 aa extracellular region which contains four IgSF domains, a 21 aa transmembrane region and a 54 aa cytoplasmic region. LAG-3 and CD4 molecules share < 20% aa sequence homology but have a similar structure (1, 2). Both molecules bind to MHC class II. LAG-3 binds to MHC class II with higher affinity compared to CD4. Both LAG-3 and CD4 genes are located on the distal part of the short arm of chromosome 12.
LAG-3 is an activation-induced molecule, expressed on activated T cells and NK cells, but not on resting T cells. Studies using LAG-3 -/- mice have shown significant delay of T cell apoptosis following antigen stimulation and increased size of memory T cells pool following infection (3, 4). It also has been reported that anti-LAG-3 antibodies up-regulate T cell activation by blocking interaction of LAG-3 and MHC class II. The study has demonstrated that LAG-3 is selectively expressed on activated CD4+CD25+ TReg cells and plays a role in their suppressive activity (5). This evidence indicated, unlike the interaction of CD4 with MHC class II that plays a positive role in T cell activation, LAG-3 binds to MHC class II and negatively regulates T cell activation through LAG-3 signaling. On the other hand, studies have shown that binding of LAG-3 to MHC class II molecules on antigen presenting cells induce maturation of dendritic cells and cytokine secretion by monocytes through MHC class II signal transduction (6). Taken together, LAG-3 may have two major functions, it negatively regulates T cells activation through LAG-3 signaling and stimulates antigen presenting cells which express MHC class II.
- Triebel, F. et al. (1990) J. Exp. Med. 171:1393.
- Baixeras, E. et al. (1992) J. Exp. Med 176:327.
- Workman, C.J. and D.A. Vignali (2003) Eur. J. Immunol. 33:970.
- Workman, C.J. et al. (2004) J. Immunol. 172:5450.
- Huang, C.T. et al. (2004) Immunity 21:503.
- Andreae, S. et al. (2003) Blood 102:2130.
Product Datasheets
Citations for Human LAG-3 PE-conjugated Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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Human T cells upregulate CD69 after coculture with xenogeneic genetically-modified pig mesenchymal stromal cells
Authors: Jiang Li, Oleg Andreyev, Man Chen, Michael Marco, Hayato Iwase, Cassandra Long et al.
Cellular Immunology
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4-1BB costimulation ameliorates T cell exhaustion induced by tonic signaling of chimeric antigen receptors
Authors: Adrienne H. Long, Waleed M. Haso, Jack F. Shern, Kelsey M. Wanhainen, Meera Murgai, Maria Ingaramo et al.
Nature Medicine
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Peripheral Blood Stem Cell Mobilization in Healthy Donors by Granulocyte Colony-Stimulating Factor Causes Preferential Mobilization of Lymphocyte Subsets
Authors: Guro Kristin Melve, Elisabeth Ersvaer, Geir Egil Eide, Einar K. Kristoffersen, Øystein Bruserud
Frontiers in Immunology
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Serpin B9 controls tumor cell killing by CAR T cells
Authors: Thomas Kimman, Anne Slomp, Anne Martens, Sarah Grabherr, Shuang Li, Eline van Diest et al.
Journal for ImmunoTherapy of Cancer
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Immune Checkpoint Profiling in Humanized Breast Cancer Mice Revealed Cell-Specific LAG-3/PD-1/TIM-3 Co-Expression and Elevated PD-1/TIM-3 Secretion
Authors: Bruss, C;Kellner, K;Albert, V;Hutchinson, JA;Seitz, S;Ortmann, O;Brockhoff, G;Wege, AK;
Cancers
Species: Mouse, Xenograft
Sample Types: Whole Cells
Applications: Flow Cytometry -
CAR T cells redirected to cell surface GRP78 display robust anti-acute myeloid leukemia activity and do not target hematopoietic progenitor cells
Authors: N Hebbar, R Epperly, A Vaidya, U Thanekar, SE Moore, M Umeda, J Ma, SL Patil, D Langfitt, S Huang, C Cheng, JM Klco, S Gottschalk, MP Velasquez
Nature Communications, 2022-01-31;13(1):587.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Responses to acute infection with SARS-CoV-2 in the lungs of rhesus macaques, baboons and marmosets
Authors: DK Singh, B Singh, SR Ganatra, M Gazi, J Cole, R Thippeshap, KJ Alfson, E Clemmons, O Gonzalez, R Escobedo, TH Lee, A Chatterjee, Y Goez-Gazi, R Sharan, M Gough, C Alvarez, A Blakley, J Ferdin, C Bartley, H Staples, L Parodi, J Callery, A Mannino, B Klaffke, P Escareno, RN Platt, V Hodara, J Scordo, S Gautam, AG Vilanova, A Olmo-Fonta, A Schami, A Oyejide, DK Ajithdoss, R Copin, A Baum, C Kyratsous, X Alvarez, M Ahmed, B Rosa, A Goodroe, J Dutton, S Hall-Urson, PA Frost, AK Voges, CN Ross, K Sayers, C Chen, C Hallam, SA Khader, M Mitreva, TJC Anderson, L Martinez-S, JL Patterson, J Turner, JB Torrelles, EJ Dick, K Brasky, LS Schlesinge, LD Giavedoni, R Carrion, D Kaushal
Nature Microbiology, 2020-12-18;6(1):73-86.
Species: Rhesus Macaca (Macaque)
Sample Types: Whole Blood
Applications: Flow Cytometry -
Interleukin-10-producing LAG3(+) regulatory T cells are associated with disease activity and abatacept treatment in rheumatoid arthritis
Authors: S Nakachi, S Sumitomo, Y Tsuchida, H Tsuchiya, M Kono, R Kato, K Sakurai, N Hanata, Y Nagafuchi, S Tateishi, H Kanda, T Okamura, K Yamamoto, K Fujio
Arthritis Res. Ther., 2017-05-16;19(1):97.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Compensatory upregulation of PD-1, LAG-3, and CTLA-4 limits the efficacy of single-agent checkpoint blockade in metastatic ovarian cancer
Authors: RY Huang, A Francois, AR McGray, A Miliotto, K Odunsi
Oncoimmunology, 2016-10-28;6(1):e1249561.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry -
Immunomodulation Induced by Stem Cell Mobilization and Harvesting in Healthy Donors: Increased Systemic Osteopontin Levels after Treatment with Granulocyte Colony-Stimulating Factor
Int J Mol Sci, 2016-07-19;17(7):.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
CD62L+ NKT cells have prolonged persistence and antitumor activity in vivo
J Clin Invest, 2016-05-16;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Cytomegalovirus-Induced Expression of CD244 after Liver Transplantation Is Associated with CD8+ T Cell Hyporesponsiveness to Alloantigen.
Authors: de Mare-Bredemeijer E, Shi X, Mancham S, van Gent R, van der Heide-Mulder M, de Boer R, Heemskerk M, de Jonge J, van der Laan L, Metselaar H, Kwekkeboom J
J Immunol, 2015-07-13;195(4):1838-48.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
TGF-beta3-expressing CD4+CD25(-)LAG3+ regulatory T cells control humoral immune responses.
Authors: Okamura T, Sumitomo S, Morita K, Iwasaki Y, Inoue M, Nakachi S, Komai T, Shoda H, Miyazaki J, Fujio K, Yamamoto K
Nat Commun, 2015-02-19;6(0):6329.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Trafficking of LAG-3 to the surface on activated T cells via its cytoplasmic domain and protein kinase C signaling.
Authors: Bae J, Lee S, Park C, Lee Y, Chun T
J Immunol, 2014-08-08;193(6):3101-12.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Human autoimmunity after lymphocyte depletion is caused by homeostatic T-cell proliferation.
Authors: Jones, Joanne L, Thompson, Sara A J, Loh, Priscill, Davies, Jessica, Tuohy, Orla C, Curry, Allison, Azzopardi, Laura, Hill-Cawthorne, Grant, Fahey, Michael, Compston, Alastair, Coles, Alasdair
Proc Natl Acad Sci U S A, 2013-11-26;110(50):20200-5.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Elevated frequencies of highly activated CD4+ T cells in HIV+ patients developing immune reconstitution inflammatory syndrome.
Authors: Antonelli LR, Mahnke Y, Hodge JN, Porter BO, Barber DL, DerSimonian R, Greenwald JH, Roby G, Mican J, Sher A, Roederer M, Sereti I
Blood, 2010-07-26;116(19):3818-27.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Different immunological effects of the molecular targeted agents sunitinib, everolimus and temsirolimus in patients with renal cell carcinoma
Authors: Yukari Kobayashi, Daisuke Yamada, Taketo Kawai, Yusuke Sato, Taro Teshima, Yuta Yamada et al.
International Journal of Oncology
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A nonhuman primate model of vertical sleeve gastrectomy facilitates mechanistic and translational research in human obesity
Authors: Julia L. Nugent, Amar Singh, Keith M. Wirth, Scott Hunter Oppler, Laura Hocum Stone, Jody L. Janecek et al.
iScience
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Mixed chimerism evolution is associated with T regulatory type 1 (Tr1) cells in a beta -thalassemic patient after haploidentical haematopoietic stem cell transplantation
Authors: Marco Andreani, Monica Emma Gianolini, Manuela Testi, MariaRosa Battarra, Galluccio Tiziana, Aldo Morrone et al.
Chimerism
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Decidual T cells exhibit a highly differentiated phenotype and demonstrate potential fetal-specificity and a strong transcriptional response to interferon
Authors: Richard M. Powell, David Lissauer, Jennifer Tamblyn, Andrew Beggs, Philip Cox, Paul Moss et al.
The Journal of Immunology
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Immunological Outcome in Haploidentical-HSC Transplanted Patients Treated with IL-10-Anergized Donor T Cells
Authors: Rosa Bacchetta, Barbarella Lucarelli, Claudia Sartirana, Silvia Gregori, Maria T. Lupo Stanghellini, Patrick Miqueu et al.
Frontiers in Immunology
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