Antibody data
- Antibody Data
- Antigen structure
- References [53]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [24]
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- Product number
- 25-0038-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD3 Monoclonal Antibody (UCHT1), PE-Cyanine7, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The UCHT1 monoclonal antibody reacts with human CD3e, a 20 kDa subunit of the TCR complex. Along with the other CD3 subunits gamma and delta, the epsilon chain is required for proper assembly, trafficking and surface expression of the TCR complex. CD3 is expressed by thymocytes in a developmentally regulated manner and by all mature T cells. Crosslinking of TCR via immobilized UCHT1 initiates an intracellular biochemical pathway resulting in cellular activation and proliferation.
- Antibody clone number
- UCHT1
- Concentration
- 5 µL/Test
Submitted references Efficient and nontoxic biomolecule delivery to primary human hematopoietic stem cells using nanostraws.
Lysine-specific demethylase 1A restricts ex vivo propagation of human HSCs and is a target of UM171.
Expansion of Group 2 Innate Lymphoid Cells in Patients with End-Stage Renal Disease and Their Clinical Significance.
Human pDCs Are Superior to cDC2s in Attracting Cytolytic Lymphocytes in Melanoma Patients Receiving DC Vaccination.
Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3.
Scalable high-throughput acoustophoresis in arrayed plastic microchannels.
Immunomodulatory Drugs in the Context of Autologous Hematopoietic Stem Cell Transplantation Associate With Reduced Pro-tumor T Cell Subsets in Multiple Myeloma.
The Genetic Polymorphisms of NLRP3 Inflammasome Associated with T Helper Cells in Patients with Multiple Myeloma.
Neisseria gonorrhoeae co-infection exacerbates vaginal HIV shedding without affecting systemic viral loads in human CD34+ engrafted mice.
EDAG promotes the expansion and survival of human CD34+ cells.
The Effects of High Mobility Group Box-1 Protein on Peripheral Treg/Th17 Balance in Patients with Atherosclerosis.
Broad CD8(+) T cell cross-recognition of distinct influenza A strains in humans.
Intact CD100-CD72 Interaction Necessary for TCR-Induced T Cell Proliferation.
ATP Release from Chemotherapy-Treated Dying Leukemia Cells Elicits an Immune Suppressive Effect by Increasing Regulatory T Cells and Tolerogenic Dendritic Cells.
Antigen receptor-redirected T cells derived from hematopoietic precursor cells lack expression of the endogenous TCR/CD3 receptor and exhibit specific antitumor capacities.
ILC2-modulated T cell-to-MDSC balance is associated with bladder cancer recurrence.
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
Frequencies of Circulating MAIT Cells Are Diminished in Chronic HCV, HIV and HCV/HIV Co-Infection and Do Not Recover during Therapy.
Follicular Regulatory CD8 T Cells Impair the Germinal Center Response in SIV and Ex Vivo HIV Infection.
Antithymocyte Globulin at Clinically Relevant Concentrations Kills Leukemic Blasts.
Early changes within the lymphocyte population are associated with the development of multiple organ dysfunction syndrome in trauma patients.
MAIT cells are activated during human viral infections.
Interaction among activated lymphocytes and mesenchymal cells through podoplanin is critical for a high IL-17 secretion.
Development of depressive symptoms post hip fracture is associated with altered immunosuppressive phenotype in regulatory T and B lymphocytes.
Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.
A truncation variant of the cation channel P2RX5 is upregulated during T cell activation.
Induction of type I and type III interferons by Borrelia burgdorferi correlates with pathogenesis and requires linear plasmid 36.
The orally available, synthetic ether lipid edelfosine inhibits T cell proliferation and induces a type I interferon response.
Depressive symptoms post hip fracture in older adults are associated with phenotypic and functional alterations in T cells.
Novel lentiviral vectors with mutated reverse transcriptase for mRNA delivery of TALE nucleases.
A comparison of DNA methylation specific droplet digital PCR (ddPCR) and real time qPCR with flow cytometry in characterizing human T cells in peripheral blood.
Age-related decrease in TCR repertoire diversity measured with deep and normalized sequence profiling.
Human mesenchymal stromal cells modulate T-cell responses through TNF-α-mediated activation of NF-κB.
Trypan blue exclusion assay by flow cytometry.
Incomplete normalization of regulatory t-cell frequency in the gut mucosa of Colombian HIV-infected patients receiving long-term antiretroviral treatment.
MHC multimer-guided and cell culture-independent isolation of functional T cell receptors from single cells facilitates TCR identification for immunotherapy.
Mother and child T cell receptor repertoires: deep profiling study.
A Novel Method Linking Antigen Presentation by Human Monocyte-Derived Macrophages to CD8(+) T Cell Polyfunctionality.
Recombinant influenza virus carrying the respiratory syncytial virus (RSV) F85-93 CTL epitope reduces RSV replication in mice.
Human neonatal naive CD4+ T cells have enhanced activation-dependent signaling regulated by the microRNA miR-181a.
HLA-DR15-derived self-peptides are involved in increased autologous T cell proliferation in multiple sclerosis.
The nucleotide sugar UDP-glucose mobilizes long-term repopulating primitive hematopoietic cells.
Ulcerative colitis impairs the acylethanolamide-based anti-inflammatory system reversal by 5-aminosalicylic acid and glucocorticoids.
HIV-induced T-cell activation/exhaustion in rectal mucosa is controlled only partially by antiretroviral treatment.
Chronically HIV-1 Infected Patients Exhibit Low Frequencies of CD25+ Regulatory T Cells.
Preserved function of regulatory T cells in chronic HIV-1 infection despite decreased numbers in blood and tissue.
Histone deacetylase inhibitors impair NK cell viability and effector functions through inhibition of activation and receptor expression.
Epigenetic biomarkers of T-cells in human glioma.
Central role of JC virus-specific CD4+ lymphocytes in progressive multi-focal leucoencephalopathy-immune reconstitution inflammatory syndrome.
Differential expression of CD300a/c on human TH1 and TH17 cells.
Human Th1 cells that express CD300a are polyfunctional and after stimulation up-regulate the T-box transcription factor eomesodermin.
Distinct roles for CCR4 and CXCR3 in the recruitment and positioning of regulatory T cells in the inflamed human liver.
Accumulation of natural killer T cells in progressive nonalcoholic fatty liver disease.
Schmiderer L, Subramaniam A, Žemaitis K, Bäckström A, Yudovich D, Soboleva S, Galeev R, Prinz CN, Larsson J, Hjort M
Proceedings of the National Academy of Sciences of the United States of America 2020 Sep 1;117(35):21267-21273
Proceedings of the National Academy of Sciences of the United States of America 2020 Sep 1;117(35):21267-21273
Lysine-specific demethylase 1A restricts ex vivo propagation of human HSCs and is a target of UM171.
Subramaniam A, Žemaitis K, Talkhoncheh MS, Yudovich D, Bäckström A, Debnath S, Chen J, Jain MV, Galeev R, Gaetani M, Zubarev RA, Larsson J
Blood 2020 Nov 5;136(19):2151-2161
Blood 2020 Nov 5;136(19):2151-2161
Expansion of Group 2 Innate Lymphoid Cells in Patients with End-Stage Renal Disease and Their Clinical Significance.
Liu GY, Deng XH, Li X, Cao YJ, Xing YF, Zhou P, Lei AH, Yang Q, Deng K, Zhang H, Zhou J
Journal of immunology (Baltimore, Md. : 1950) 2020 Jul 1;205(1):36-44
Journal of immunology (Baltimore, Md. : 1950) 2020 Jul 1;205(1):36-44
Human pDCs Are Superior to cDC2s in Attracting Cytolytic Lymphocytes in Melanoma Patients Receiving DC Vaccination.
van Beek JJP, Flórez-Grau G, Gorris MAJ, Mathan TSM, Schreibelt G, Bol KF, Textor J, de Vries IJM
Cell reports 2020 Jan 28;30(4):1027-1038.e4
Cell reports 2020 Jan 28;30(4):1027-1038.e4
Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3.
Trovato R, Fiore A, Sartori S, Canè S, Giugno R, Cascione L, Paiella S, Salvia R, De Sanctis F, Poffe O, Anselmi C, Hofer F, Sartoris S, Piro G, Carbone C, Corbo V, Lawlor R, Solito S, Pinton L, Mandruzzato S, Bassi C, Scarpa A, Bronte V, Ugel S
Journal for immunotherapy of cancer 2019 Sep 18;7(1):255
Journal for immunotherapy of cancer 2019 Sep 18;7(1):255
Scalable high-throughput acoustophoresis in arrayed plastic microchannels.
Dubay R, Lissandrello C, Swierk P, Moore N, Doty D, Fiering J
Biomicrofluidics 2019 May;13(3):034105
Biomicrofluidics 2019 May;13(3):034105
Immunomodulatory Drugs in the Context of Autologous Hematopoietic Stem Cell Transplantation Associate With Reduced Pro-tumor T Cell Subsets in Multiple Myeloma.
Di Lullo G, Marcatti M, Heltai S, Tresoldi C, Paganoni AM, Bordignon C, Ciceri F, Protti MP
Frontiers in immunology 2018;9:3171
Frontiers in immunology 2018;9:3171
The Genetic Polymorphisms of NLRP3 Inflammasome Associated with T Helper Cells in Patients with Multiple Myeloma.
Zhao X, Hua M, Yan S, Yu J, Han F, Zhong C, Wang R, Zhang C, Hou M, Ma D
Journal of immunology research 2018;2018:7569809
Journal of immunology research 2018;2018:7569809
Neisseria gonorrhoeae co-infection exacerbates vaginal HIV shedding without affecting systemic viral loads in human CD34+ engrafted mice.
Xu SX, Leontyev D, Kaul R, Gray-Owen SD
PloS one 2018;13(1):e0191672
PloS one 2018;13(1):e0191672
EDAG promotes the expansion and survival of human CD34+ cells.
Zhao K, Zheng WW, Dong XM, Yin RH, Gao R, Li X, Liu JF, Zhan YQ, Yu M, Chen H, Ge CH, Ning HM, Yang XM, Li CY
PloS one 2018;13(1):e0190794
PloS one 2018;13(1):e0190794
The Effects of High Mobility Group Box-1 Protein on Peripheral Treg/Th17 Balance in Patients with Atherosclerosis.
Ding JW, Zhou T, Zheng XX, Wang XA, Tong XH, Luo CY, Zhang ZQ, Yu B
Acta Cardiologica Sinica 2018 Sep;34(5):399-408
Acta Cardiologica Sinica 2018 Sep;34(5):399-408
Broad CD8(+) T cell cross-recognition of distinct influenza A strains in humans.
Grant EJ, Josephs TM, Loh L, Clemens EB, Sant S, Bharadwaj M, Chen W, Rossjohn J, Gras S, Kedzierska K
Nature communications 2018 Dec 21;9(1):5427
Nature communications 2018 Dec 21;9(1):5427
Intact CD100-CD72 Interaction Necessary for TCR-Induced T Cell Proliferation.
Jiang X, Björkström NK, Melum E
Frontiers in immunology 2017;8:765
Frontiers in immunology 2017;8:765
ATP Release from Chemotherapy-Treated Dying Leukemia Cells Elicits an Immune Suppressive Effect by Increasing Regulatory T Cells and Tolerogenic Dendritic Cells.
Lecciso M, Ocadlikova D, Sangaletti S, Trabanelli S, De Marchi E, Orioli E, Pegoraro A, Portararo P, Jandus C, Bontadini A, Redavid A, Salvestrini V, Romero P, Colombo MP, Di Virgilio F, Cavo M, Adinolfi E, Curti A
Frontiers in immunology 2017;8:1918
Frontiers in immunology 2017;8:1918
Antigen receptor-redirected T cells derived from hematopoietic precursor cells lack expression of the endogenous TCR/CD3 receptor and exhibit specific antitumor capacities.
Van Caeneghem Y, De Munter S, Tieppo P, Goetgeluk G, Weening K, Verstichel G, Bonte S, Taghon T, Leclercq G, Kerre T, Debets R, Vermijlen D, Abken H, Vandekerckhove B
Oncoimmunology 2017;6(3):e1283460
Oncoimmunology 2017;6(3):e1283460
ILC2-modulated T cell-to-MDSC balance is associated with bladder cancer recurrence.
Chevalier MF, Trabanelli S, Racle J, Salomé B, Cesson V, Gharbi D, Bohner P, Domingos-Pereira S, Dartiguenave F, Fritschi AS, Speiser DE, Rentsch CA, Gfeller D, Jichlinski P, Nardelli-Haefliger D, Jandus C, Derré L
The Journal of clinical investigation 2017 Aug 1;127(8):2916-2929
The Journal of clinical investigation 2017 Aug 1;127(8):2916-2929
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
Su S, Liao J, Liu J, Huang D, He C, Chen F, Yang L, Wu W, Chen J, Lin L, Zeng Y, Ouyang N, Cui X, Yao H, Su F, Huang JD, Lieberman J, Liu Q, Song E
Cell research 2017 Apr;27(4):461-482
Cell research 2017 Apr;27(4):461-482
Frequencies of Circulating MAIT Cells Are Diminished in Chronic HCV, HIV and HCV/HIV Co-Infection and Do Not Recover during Therapy.
Spaan M, Hullegie SJ, Beudeker BJ, Kreefft K, van Oord GW, Groothuismink ZM, van Tilborg M, Rijnders B, de Knegt RJ, Claassen MA, Boonstra A
PloS one 2016;11(7):e0159243
PloS one 2016;11(7):e0159243
Follicular Regulatory CD8 T Cells Impair the Germinal Center Response in SIV and Ex Vivo HIV Infection.
Miles B, Miller SM, Folkvord JM, Levy DN, Rakasz EG, Skinner PJ, Connick E
PLoS pathogens 2016 Oct;12(10):e1005924
PLoS pathogens 2016 Oct;12(10):e1005924
Antithymocyte Globulin at Clinically Relevant Concentrations Kills Leukemic Blasts.
Dabas R, Lee R, Servito MT, Dharmani-Khan P, Modi M, van Slyke T, Luider J, Durand C, Larratt L, Brandwein J, Morris D, Daly A, Khan FM, Storek J
Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation 2016 May;22(5):815-24
Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation 2016 May;22(5):815-24
Early changes within the lymphocyte population are associated with the development of multiple organ dysfunction syndrome in trauma patients.
Manson J, Cole E, De'Ath HD, Vulliamy P, Meier U, Pennington D, Brohi K
Critical care (London, England) 2016 Jun 7;20(1):176
Critical care (London, England) 2016 Jun 7;20(1):176
MAIT cells are activated during human viral infections.
van Wilgenburg B, Scherwitzl I, Hutchinson EC, Leng T, Kurioka A, Kulicke C, de Lara C, Cole S, Vasanawathana S, Limpitikul W, Malasit P, Young D, Denney L, STOP-HCV consortium, Moore MD, Fabris P, Giordani MT, Oo YH, Laidlaw SM, Dustin LB, Ho LP, Thompson FM, Ramamurthy N, Mongkolsapaya J, Willberg CB, Screaton GR, Klenerman P
Nature communications 2016 Jun 23;7:11653
Nature communications 2016 Jun 23;7:11653
Interaction among activated lymphocytes and mesenchymal cells through podoplanin is critical for a high IL-17 secretion.
Noack M, Ndongo-Thiam N, Miossec P
Arthritis research & therapy 2016 Jun 23;18:148
Arthritis research & therapy 2016 Jun 23;18:148
Development of depressive symptoms post hip fracture is associated with altered immunosuppressive phenotype in regulatory T and B lymphocytes.
Duggal NA, Upton J, Phillips AC, Lord JM
Biogerontology 2016 Feb;17(1):229-39
Biogerontology 2016 Feb;17(1):229-39
Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.
Miles B, Miller SM, Folkvord JM, Kimball A, Chamanian M, Meditz AL, Arends T, McCarter MD, Levy DN, Rakasz EG, Skinner PJ, Connick E
Nature communications 2015 Oct 20;6:8608
Nature communications 2015 Oct 20;6:8608
A truncation variant of the cation channel P2RX5 is upregulated during T cell activation.
Abramowski P, Ogrodowczyk C, Martin R, Pongs O
PloS one 2014;9(9):e104692
PloS one 2014;9(9):e104692
Induction of type I and type III interferons by Borrelia burgdorferi correlates with pathogenesis and requires linear plasmid 36.
Krupna-Gaylord MA, Liveris D, Love AC, Wormser GP, Schwartz I, Petzke MM
PloS one 2014;9(6):e100174
PloS one 2014;9(6):e100174
The orally available, synthetic ether lipid edelfosine inhibits T cell proliferation and induces a type I interferon response.
Abramowski P, Otto B, Martin R
PloS one 2014;9(3):e91970
PloS one 2014;9(3):e91970
Depressive symptoms post hip fracture in older adults are associated with phenotypic and functional alterations in T cells.
Duggal NA, Upton J, Phillips AC, Hampson P, Lord JM
Immunity & ageing : I & A 2014;11(1):25
Immunity & ageing : I & A 2014;11(1):25
Novel lentiviral vectors with mutated reverse transcriptase for mRNA delivery of TALE nucleases.
Mock U, Riecken K, Berdien B, Qasim W, Chan E, Cathomen T, Fehse B
Scientific reports 2014 Sep 18;4:6409
Scientific reports 2014 Sep 18;4:6409
A comparison of DNA methylation specific droplet digital PCR (ddPCR) and real time qPCR with flow cytometry in characterizing human T cells in peripheral blood.
Wiencke JK, Bracci PM, Hsuang G, Zheng S, Hansen H, Wrensch MR, Rice T, Eliot M, Kelsey KT
Epigenetics 2014 Oct;9(10):1360-5
Epigenetics 2014 Oct;9(10):1360-5
Age-related decrease in TCR repertoire diversity measured with deep and normalized sequence profiling.
Britanova OV, Putintseva EV, Shugay M, Merzlyak EM, Turchaninova MA, Staroverov DB, Bolotin DA, Lukyanov S, Bogdanova EA, Mamedov IZ, Lebedev YB, Chudakov DM
Journal of immunology (Baltimore, Md. : 1950) 2014 Mar 15;192(6):2689-98
Journal of immunology (Baltimore, Md. : 1950) 2014 Mar 15;192(6):2689-98
Human mesenchymal stromal cells modulate T-cell responses through TNF-α-mediated activation of NF-κB.
Dorronsoro A, Ferrin I, Salcedo JM, Jakobsson E, Fernández-Rueda J, Lang V, Sepulveda P, Fechter K, Pennington D, Trigueros C
European journal of immunology 2014 Feb;44(2):480-8
European journal of immunology 2014 Feb;44(2):480-8
Trypan blue exclusion assay by flow cytometry.
Avelar-Freitas BA, Almeida VG, Pinto MC, Mourão FA, Massensini AR, Martins-Filho OA, Rocha-Vieira E, Brito-Melo GE
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas 2014 Apr;47(4):307-15
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas 2014 Apr;47(4):307-15
Incomplete normalization of regulatory t-cell frequency in the gut mucosa of Colombian HIV-infected patients receiving long-term antiretroviral treatment.
Rueda CM, Velilla PA, Chougnet CA, Rugeles MT
PloS one 2013;8(8):e71062
PloS one 2013;8(8):e71062
MHC multimer-guided and cell culture-independent isolation of functional T cell receptors from single cells facilitates TCR identification for immunotherapy.
Dössinger G, Bunse M, Bet J, Albrecht J, Paszkiewicz PJ, Weißbrich B, Schiedewitz I, Henkel L, Schiemann M, Neuenhahn M, Uckert W, Busch DH
PloS one 2013;8(4):e61384
PloS one 2013;8(4):e61384
Mother and child T cell receptor repertoires: deep profiling study.
Putintseva EV, Britanova OV, Staroverov DB, Merzlyak EM, Turchaninova MA, Shugay M, Bolotin DA, Pogorelyy MV, Mamedov IZ, Bobrynina V, Maschan M, Lebedev YB, Chudakov DM
Frontiers in immunology 2013;4:463
Frontiers in immunology 2013;4:463
A Novel Method Linking Antigen Presentation by Human Monocyte-Derived Macrophages to CD8(+) T Cell Polyfunctionality.
Short KR, Grant EJ, Vissers M, Reading PC, Diavatopoulos DA, Kedzierska K
Frontiers in immunology 2013;4:389
Frontiers in immunology 2013;4:389
Recombinant influenza virus carrying the respiratory syncytial virus (RSV) F85-93 CTL epitope reduces RSV replication in mice.
De Baets S, Schepens B, Sedeyn K, Schotsaert M, Roose K, Bogaert P, Fiers W, Saelens X
Journal of virology 2013 Mar;87(6):3314-23
Journal of virology 2013 Mar;87(6):3314-23
Human neonatal naive CD4+ T cells have enhanced activation-dependent signaling regulated by the microRNA miR-181a.
Palin AC, Ramachandran V, Acharya S, Lewis DB
Journal of immunology (Baltimore, Md. : 1950) 2013 Mar 15;190(6):2682-91
Journal of immunology (Baltimore, Md. : 1950) 2013 Mar 15;190(6):2682-91
HLA-DR15-derived self-peptides are involved in increased autologous T cell proliferation in multiple sclerosis.
Mohme M, Hotz C, Stevanovic S, Binder T, Lee JH, Okoniewski M, Eiermann T, Sospedra M, Rammensee HG, Martin R
Brain : a journal of neurology 2013 Jun;136(Pt 6):1783-98
Brain : a journal of neurology 2013 Jun;136(Pt 6):1783-98
The nucleotide sugar UDP-glucose mobilizes long-term repopulating primitive hematopoietic cells.
Kook S, Cho J, Lee SB, Lee BC
The Journal of clinical investigation 2013 Aug;123(8):3420-35
The Journal of clinical investigation 2013 Aug;123(8):3420-35
Ulcerative colitis impairs the acylethanolamide-based anti-inflammatory system reversal by 5-aminosalicylic acid and glucocorticoids.
Suárez J, Romero-Zerbo Y, Márquez L, Rivera P, Iglesias M, Bermúdez-Silva FJ, Andreu M, Rodríguez de Fonseca F
PloS one 2012;7(5):e37729
PloS one 2012;7(5):e37729
HIV-induced T-cell activation/exhaustion in rectal mucosa is controlled only partially by antiretroviral treatment.
Rueda CM, Velilla PA, Chougnet CA, Montoya CJ, Rugeles MT
PloS one 2012;7(1):e30307
PloS one 2012;7(1):e30307
Chronically HIV-1 Infected Patients Exhibit Low Frequencies of CD25+ Regulatory T Cells.
Rios CM, Velilla PA, Rugeles MT
The open virology journal 2012;6:49-58
The open virology journal 2012;6:49-58
Preserved function of regulatory T cells in chronic HIV-1 infection despite decreased numbers in blood and tissue.
Angin M, Kwon DS, Streeck H, Wen F, King M, Rezai A, Law K, Hongo TC, Pyo A, Piechocka-Trocha A, Toth I, Pereyra F, Ghebremichael M, Rodig SJ, Milner DA Jr, Richter JM, Altfeld M, Kaufmann DE, Walker BD, Addo MM
The Journal of infectious diseases 2012 May 15;205(10):1495-500
The Journal of infectious diseases 2012 May 15;205(10):1495-500
Histone deacetylase inhibitors impair NK cell viability and effector functions through inhibition of activation and receptor expression.
Rossi LE, Avila DE, Spallanzani RG, Ziblat A, Fuertes MB, Lapyckyj L, Croci DO, Rabinovich GA, Domaica CI, Zwirner NW
Journal of leukocyte biology 2012 Feb;91(2):321-31
Journal of leukocyte biology 2012 Feb;91(2):321-31
Epigenetic biomarkers of T-cells in human glioma.
Wiencke JK, Accomando WP, Zheng S, Patoka J, Dou X, Phillips JJ, Hsuang G, Christensen BC, Houseman EA, Koestler DC, Bracci P, Wiemels JL, Wrensch M, Nelson HH, Kelsey KT
Epigenetics 2012 Dec 1;7(12):1391-402
Epigenetics 2012 Dec 1;7(12):1391-402
Central role of JC virus-specific CD4+ lymphocytes in progressive multi-focal leucoencephalopathy-immune reconstitution inflammatory syndrome.
Aly L, Yousef S, Schippling S, Jelcic I, Breiden P, Matschke J, Schulz R, Bofill-Mas S, Jones L, Demina V, Linnebank M, Ogg G, Girones R, Weber T, Sospedra M, Martin R
Brain : a journal of neurology 2011 Sep;134(Pt 9):2687-702
Brain : a journal of neurology 2011 Sep;134(Pt 9):2687-702
Differential expression of CD300a/c on human TH1 and TH17 cells.
Simhadri VR, Mariano JL, Zhou Q, DeBell KE, Borrego F
BMC immunology 2011 Nov 2;12:62
BMC immunology 2011 Nov 2;12:62
Human Th1 cells that express CD300a are polyfunctional and after stimulation up-regulate the T-box transcription factor eomesodermin.
Narayanan S, Silva R, Peruzzi G, Alvarez Y, Simhadri VR, Debell K, Coligan JE, Borrego F
PloS one 2010 May 13;5(5):e10636
PloS one 2010 May 13;5(5):e10636
Distinct roles for CCR4 and CXCR3 in the recruitment and positioning of regulatory T cells in the inflamed human liver.
Oo YH, Weston CJ, Lalor PF, Curbishley SM, Withers DR, Reynolds GM, Shetty S, Harki J, Shaw JC, Eksteen B, Hubscher SG, Walker LS, Adams DH
Journal of immunology (Baltimore, Md. : 1950) 2010 Mar 15;184(6):2886-98
Journal of immunology (Baltimore, Md. : 1950) 2010 Mar 15;184(6):2886-98
Accumulation of natural killer T cells in progressive nonalcoholic fatty liver disease.
Syn WK, Oo YH, Pereira TA, Karaca GF, Jung Y, Omenetti A, Witek RP, Choi SS, Guy CD, Fearing CM, Teaberry V, Pereira FE, Adams DH, Diehl AM
Hepatology (Baltimore, Md.) 2010 Jun;51(6):1998-2007
Hepatology (Baltimore, Md.) 2010 Jun;51(6):1998-2007
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Supportive validation
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- Staining of human peripheral blood mononuclear cells with CD45 Pacific Blue, CD19 FITC and CD3 PE-Cyanine5. As expected based on known relative expression patterns, CD3 clone UCHT1 stains a subset of lymphocytes (pink), but not monocytes (orange) and granulocytes (blue).
Supportive validation
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- Figure 6 Profile of T-lymphocytes stained with monoclonal antibody anti-CD3-FITCfollowed by treatment with propidium iodide (PI) and trypan blue (TB) at 0.002and 0.4% (w/v) or PBS (untreated control).
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- Figure 7 A , Percentage of dead human T-lymphocytes(CD3-FITC + cells) submitted to cell culture at temperatures (T)of 37degC (physiologic temperature) or 50degC (high-stress temperature) followed bystaining with trypan blue (TB) or propidium iodide (PI). B ,Dot-plot graph profile between human lymphocytes submitted to pretreatment withhigh-stress temperature (50degC) followed by staining with TB and PI andmonoclonal antibody anti-CD3-FITC + . C , Pearson'scorrelation test between dead CD3 + lymphocytes using PI and TB flowcytometry assays.
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- Figure 3 Functional analysis of NRTLV-delivered TALEN constructs. (a) Schematic vector design of I: LeGO-iTALEN-iG2 (wPRE), II: LeGO-iTALEN-iG2-wPRE-BGH-p(A) and III: LeGO-iTALEN-iG2-wPRE-SV40-p(A) 3 rd generation LVV derived from LeGO-system 11 . CMV = CMV-ie promoter; Delta (DeltaU3), R, U5 = elements of SIN-LTR, self-inactivating long terminal repeat; Psi = Psi , packaging signal; RRE = Rev response element; SFFV = promoter of spleen focus-forming virus; wPRE = Woodchuck hepatitis virus posttranscriptional regulatory element; IRES = internal ribosome entry site; eGFP = enhanced green fluorescent protein; p(A) = polyadenylation signal; BGH = bovine growth hormone; SV40 = simian virus 40. (b) Knockout of CCR5 in reporter cell line CCR5+/293T-cell clones were co-transduced with non-concentrated NRTLVs delivering different iTALEN-constructs with either no internal polyadenylation (p(A)) signal (iTALEN-wPRE), or internal BGH-p(A) (iTALEN-wPRE-BGH-p(A)) or SV40-p(A) (iTALEN-wPRE-SV40-p(A)) signals downstream of the wPRE-element, respectively. Mock-transduction and transduction of left or right TALEN-arms, only, served as negative controls (homodimers only). Measured for 3 independently produced vector preparations, each time in duplicates, *p
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- Figure 2 Single-cell PCR delivers sequences of functional CMV-specific TCRs. (A) PBMCs from Donor 1 were recovered and stained with HLA-B7/pp65 417-426 multimers. Dot plot shows the further analyzed CD8 and MHC multimer double-positive cell population. Cells were pre-gated on live lymphocytes (propidium iodide-negative, and CD3-positive). (B) TCR SCAN as described in Figure 1 and agarose gel electrophoresis of the resulting PCR products was performed. The photography shows the agarose gel. Upper row shows alpha-chain products matched with the respective beta-chains in the lower row. White boxes indicate alpha- and matched beta-chain-products derived from identical single cell samples. (C) The table summarizes the V- D- J- segment type and amino acid sequences of TCRs identified from CMV-multimer positive T cells in Figure 2A . TCR1A was identified 11 times and TCR 1B was detected once. (D) MHC multimer-positive T cells from the same donor were in vitro expanded and six T cell clones were successfully maintained. All clones contained TCR1A as confirmed by PCR and sequencing. The left FACS plot shows HLA-B7/pp65 417-426 staining and the right FACS plot shows staining with an irrelevant MHC multimer. (E) PBMCs from donor 2 were recovered and stained with HLA-B8/IE-1 199-207 multimers. The dot plot shows the further analyzed CD8 and MHC multimer double-positive cell population. Cells were pre-gated on living lymphocytes (propidium iodide negative and CD3 positive). (F) TCR SCAN a
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- Figure 3 Characterization of a small diverse T cell repertoire and transgenic expression of detected TCRs. (A) PBMCs from donor 3 were stained with HLA-B8/IE-1 88-96 multimers. Dot plot shows the further analyzed CD8 and MHC multimer double-positive cell population. Cells were pre-gated on living lymphocytes (propidium iodide-negative and CD3-positive). (B) A PCR slide with single antigen-specific T cells from Figure 3A were FACS-isolated. TCR SCAN as described in Figure 1 and agarose gel electrophoresis of the resulting PCR products was performed. Upper row shows alpha-chain products matched with the respective beta-chains in the lower row. White boxes indicate samples alpha- and matched beta-chain-products derived from identical single cell samples. (C) The table summarizes the V- D- J- segment type and amino acid sequences of TCRs identified from CMV-multimer positive T cells in Figure 3A . In three independent experiments we identified nine different TCRs (TCR 3A-I) (D) Pie chart indicates the prevalence of identified TCRs from donor 3. Percentages represent incidence of respective TCR divided by total number of positive samples. (E) Sequences from TCR 3D, 3E and 3G were expressed in Jurkat76 T cells by retroviral gene transfer. Non-transduced (left FACS plot) and transduced Jurkat76 T cells were analyzed for expression of CD3 and MHC multimer binding.
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- Figure 5 lp36 contributes to the association of B. burgdorferi with specific populations of dendritic cells. Human PBMCs (4x10 6 ) were co-incubated with 4x10 7 GFP-tagged B31 (black bars), A3-M9 lp36- (white bars) or A3-M9 lp36-/lp36+ (cross-hatched bars) B. burgdorferi for 6 hours at 4degC or 37degC. The percentages of GFP + mDC1s (CD19 - CD3 - BDCA2 - BDCA1 + ) ( A ), pDCs (CD19 - CD3 - BDCA2 + BDCA1 - ) or ( B ) mDC2s (CD19 - CD3 - BDCA3 + BDCA2 - ) ( C ) were determined by multiparameter flow cytometry. Dot plots representing 500,000 collected events are provided to illustrate gating strategies (left). Column graphs represent the mean and standard deviation of three biological replicates (right). Statistical analysis was performed using a one-way ANOVA with a Tukey's post-test for multiple comparisons.
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- Figure 2 T FR expansion in lymphoid tissues during chronic SIV infection. ( a ) Disaggregated lymph node and spleen cells from SIV uninfected ( n =9) or chronically SIV-infected rhesus macaques ( n =11) were analysed by flow cytometry. Representative examples of flow cytometry gating are shown. Of viable CD3 + CD8 - cells, follicular subsets were defined as CXCR5 + cells (F) and germinal centre subsets were defined as CXCR5 hi PD-1 hi cells (GC). Of these subsets, regulatory cells were defined as CD25 hi CD127 - . T FR (CXCR5 + CD25 hi CD127 - ) were Foxp3 + , whereas T FH (CXCR5 + CD25 lo/- ) were Foxp3 - . ( b ) The percentages of each rhesus macaque regulatory subset, as analysed in a are shown. ( c ) The ratios of each regulatory cell population to its non-regulatory cell counterpart are shown. ( d ) The percentage of total CTLA-4 expression is shown in SIV-uninfected ( n =9) and chronically SIV-infected ( n =8) rhesus macaques. The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Mann-Whitney (Wilcoxon) tests to compare unpaired, nonparametric values and significance is denoted by asterisks where * P
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- Figure 3 HIV entry and replication promote T FR expansion. Disaggregated tonsil cells were spinoculated with X4 or R5 HIV and T FR populations were analysed by flow cytometry ( n =15). ( a ) A representative example of tonsil cell flow gating. From viable CD3 + CD8 - cells, T FR are defined as CXCR5 + and CD25 hi CD127 - . T FR cells contain Foxp3 + cells, whereas remaining T FH (CXCR5 + CD25 lo/- ) cells are Foxp3 - . ( b ) Percentages of T FR determined by gating strategies in a are shown. Experimental conditions include mock-spinoculated cells cultured with PMA (50 ng ml -1 ) and ionomycin (1 mug ml -1 ) or exogenous TGF-beta (100 ng ml -1 ) for 24 h and cells pretreated to block CXCR4 (AMD, 200 muM) and CCR5 (MVC, 2 muM). ( c ) Using flow cytometry counting beads, the number of cells per mul were determined for total (CD3 + CD8 - ), T FH (CXCR5 + CD25 lo/- ) and T FR (CXCR5 + CD25 hi CD127 - ) subsets in mock- and X4-spinoculated samples ( n =3). ( d ) Bcl-6 expression is shown in CXCR5- (grey), T FH (blue) and T FR (red) populations after mock-, X4- or R5-spinoculation ( n =5). ( e ) Blimp-1 expression was also determined as in d . The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Friedman nonparametric tests ( b , d , e ) and significance is denoted by asterisks where * P
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- Figure 3. Phenotype and endogenous TCR expression of CD34 + HPC-derived transgenic AR + T cells. Flow cytometric analysis of the AR-transgenic T cells. (A) CAR-transgenic GFP + cells of cultures transduced to express either the CAR:zeta or the CAR:28zeta were analyzed on day 26 of OP9-DL1 culture for CD3 and TCRalphabeta expression. As a control, GFP - cells are shown from the OP9-DL1 culture transduced to express the CAR:zeta ( N = 5). (B) Dot plots show CD3 expression of cells from the OP9-DL1 cultures transgenic for the wtTCR, TCR:zeta and TCR:28zeta. Vbeta14 staining is used to mark transgene expression, as no GFP is expressed by the transgenic cells ( N = 5). (C) Surface and cytoplasmic staining for CD3 of in vitro generated mature T cells that were expanded for one cycle on feeder cells in the presence of cytokines. (D) Expression of various membrane markers by the CD27 + CD1a - mature T cells at the end of OP9-DL1 culture (46 d) ( N = 2). (E) Day 0: fresh cord blood after MACS CD34 enrichment sorted using the sorting window shown. Day 13: cord blood cells cultured on OP9-DL1 were sorted for CD5 CD7 double positive cells, using the indicated sorting window. The cells were then transduced to express CAR:28zeta and further differentiated on OP9-DL1 feeder layer. Day 21: analysis of the transgenic GFP + cultured cells for DP cells and CD27 + CD1a - mature cells. (F) Flow cytometric analysis of GFP + CAR:28zeta-transgenic cultures, gated on GFP + CD27 + CD1a - mature AR + c
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- Figure 3 Naive CD4 + T cells are converted to functional Tregs by tumor-infiltrating DCs and tumor conditioned medium (CM). (A-C) Naive CD4 + T cells from peripheral blood of patients with invasive breast carcinoma were co-cultured with or without autologous pDCs isolated from tumor (TI) or peripheral blood (PB) for 9 days in the presence or absence of 30% CM from autologous tumor slices or adjacent normal tissue slices. (A , B) Non-adherent cells from co-cultures were stained for CD3, CD4, CD25 and intracellular Foxp3, and analyzed by flow cytometry. Representative plots of gated CD3 + CD4 + cells (A) and quantification of percentage of Foxp3 + CD25 + cells among CD3 + CD4 + cells (B) are shown (mean +- SEM, n = 19; * P < 0.05, ** P < 0.01, *** P < 0.001 by Student's t -test). (C) Expression of Treg-associated genes, assessed by qRT-PCR normalized to GAPDH , in sorted CD4 + T cells, relative to expression in cultures without DCs or CM (mean +- SEM, n = 19; * P < 0.05, ** P < 0.01, *** P < 0.001 compared with naive CD4 + T cells cultured alone by Student's t -test). (D-G) Effect of naive CD4 + T cell-derived Tregs, obtained by co-culture with TI pDCs and tumor CM as above, on function of autologous tumor-specific CD8 + T cells. Tumor-specific CD8 + T cells were generated for each subject by stimulating autologous PB CD8 + T cells with autologous tumor lysate-pulsed autologous DCs. Tregs were recovered from co-cultures by magnetic sorting. (D) CFSE-labeled CD8 + T ce
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- Figure 6 In vivo knockdown of PITPNM3 in CD4 + T cells reverses immunosuppression and inhibits tumor progression in humanized mice. (A) Humanized mice bearing palpable MDA-MB-231 orthotopic xenografts were intraperitoneally injected daily for 14 days with PBS, 1 nmol CD4-aptamer-control siRNA (AsiC-con) or CD4-aptamer-siRNA targeting PITPNM3 (sequence in A , AsiC-PI) to assess the role of PITPNM3 in TI Tregs, and other T cells and tumor control. Experimental schematic is provided in Supplementary information, Figure S9A . (B) Representative immunoblots showing selective knockdown of PITPNM3 protein in PB CD4 + T cells, but not tumor xenografts ( n = 3). (C) PITPNM3 knockdown did not affect the distribution of human CD45 + hematopoietic cells, CD4 + and CD8 + T cells, and CD14 + monocytes in the peripheral blood of humanized mice. Representative flow plots are shown ( n = 3). (D , E) Effect of PITPNM3 knockdown on TI naive CD4 + , Tregs and CD8 + T cell numbers, and apoptosis by TUNEL assay in xenografts. D shows representative immunofluorescence microscopy images. Top row indicates CD4 + naive T cells by arrows; the second row indicates CD4 + CD45RO + Foxp3 - CD4 + memory T cells (yellow arrows) and Foxp3 + Tregs (white arrows). Scale bar, 50 mum. E shows number of cells of each subtype/high power field in eight mice ( ** P < 0.01, *** P < 0.001 compared to PBS group by Student's t -test). (F) Flow cytometry analysis of gated human CD3 + CD4 + cells isolated from xenogra
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- Figure 7 CD4-aptamer-siRNA targeting PITPNM3 reduces TI Tregs and inhibits tumor progression in humanized mice with circulating human Tregs. Humanized mice, implanted with MDA-MB-231 tumors and concurrently injected intravenously with autologous Tregs, were intraperitoneally injected daily for 14 days after tumors became palpable with PBS, 1 nmol CD4-aptamer-control siRNA (AsiC-con) or CD4-aptamer-siRNA targeting PITPNM3 to assess the role of PITPNM3 in TI Tregs, and other T cells and tumor control. Tregs were administered every 10 days after the initial injection and mice were sacrificed 30 days after tumor cell inoculation. (A) Experimental schematic. (B , C) Peripheral blood cells of humanized mice were stained for human CD3, CD4 and Foxp3, and analyzed by flow cytometry. A representative flow plot (B) and the percentage (mean +- SEM) of PB CD4 + cells that are CFSE + Tregs in six mice per group (C) are shown. (D , E) Isolated cells from xenografts were stained for human CD3, CD4 and Foxp3. The percentage (mean +- SEM) of six mice per group (D) and representative flow plot (E) of FoxP3 + Tregs are shown. Most Tregs were CFSE - (i.e., did not come from infused Tregs) and the number of TI Tregs was reduced by knocking down PITPNM3 in CD4 + T cells ( *** P < 0.001 compared to the PBS group by Student's t -test). (F) Tumor size (mean +- SEM, n = 6 per group; *** P < 0.001 by two-way ANOVA with Bonferroni multiple comparison tests). (G) Lung metastases assessed by qRT-PCR