Antibody data
- Antibody Data
- Antigen structure
- References [48]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [38]
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- Product number
- 11-0038-80 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD3 Monoclonal Antibody (UCHT1), FITC, 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. Applications Reported: The UCHT1 antibody has been reported for use in flow cytometric analysis. Applications Tested: This UCHT1 antibody has been tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Excitation: 488 nm; Emission: 520 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Conjugate
- Green dye
- Isotype
- IgG
- Antibody clone number
- UCHT1
- Vial size
- 25 µg
- Concentration
- 0.5 mg/mL
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references IL-36 cytokines imprint a colitogenic phenotype on CD4(+) T helper cells.
Aberrant gut-microbiota-immune-brain axis development in premature neonates with brain damage.
Single-cell RNA sequencing reveals ex vivo signatures of SARS-CoV-2-reactive T cells through 'reverse phenotyping'.
Dendritic Cell Maturation Regulates TSPAN7 Function in HIV-1 Transfer to CD4(+) T Lymphocytes.
Single-cell transcriptomics reveals regulators underlying immune cell diversity and immune subtypes associated with prognosis in nasopharyngeal carcinoma.
Aging affects responsiveness of peripheral blood mononuclear cells to immunosuppression of periodontal ligament stem cells.
Involvement of natural killer cells in the pathogenesis of endometriosis in patients with pelvic pain.
Th2-like T Follicular Helper Cells Promote Functional Antibody Production during Plasmodium falciparum Infection.
Skewed T cell responses to Epstein-Barr virus in long-term asymptomatic kidney transplant recipients.
Inflammation induced by incomplete radiofrequency ablation accelerates tumor progression and hinders PD-1 immunotherapy.
Loss of ARPC1B impairs cytotoxic T lymphocyte maintenance and cytolytic activity.
Biallelic mutations in DNA ligase 1 underlie a spectrum of immune deficiencies.
Antigen receptor-redirected T cells derived from hematopoietic precursor cells lack expression of the endogenous TCR/CD3 receptor and exhibit specific antitumor capacities.
Follicular Regulatory T Cells Are Highly Permissive to R5-Tropic HIV-1.
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
The 4E-BP-eIF4E axis promotes rapamycin-sensitive growth and proliferation in lymphocytes.
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.
Dendritic cell-activated cytokine-induced killer cell-mediated immunotherapy is safe and effective for cancer patients >65 years old.
Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.
Characterization of the in vivo immune network of IDO, tryptophan metabolism, PD-L1, and CTLA-4 in circulating immune cells in melanoma.
Clinical significance of plasmacytoid dendritic cells and myeloid-derived suppressor cells in melanoma.
Serum macrophage-derived chemokine/CCL22 levels are associated with glioma risk, CD4 T cell lymphopenia and survival time.
Induction of type I and type III interferons by Borrelia burgdorferi correlates with pathogenesis and requires linear plasmid 36.
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.
Human mesenchymal stromal cells modulate T-cell responses through TNF-α-mediated activation of NF-κB.
Trypan blue exclusion assay by flow cytometry.
HER2 as a promising target for cytotoxicity T cells in human melanoma therapy.
MHC multimer-guided and cell culture-independent isolation of functional T cell receptors from single cells facilitates TCR identification for immunotherapy.
Evaluation of clinical and immunological responses: a 2-year follow-up study in children with allergic rhinitis due to house dust mite.
Recombinant influenza virus carrying the respiratory syncytial virus (RSV) F85-93 CTL epitope reduces RSV replication in mice.
Inhibition of CXCR3-mediated chemotaxis by the human chemokine receptor-like protein CCX-CKR.
Association of a NOD2 gene polymorphism and T-helper 17 cells with presumed ocular toxoplasmosis.
Assessment of immune interference, antagonism, and diversion following human immunization with biallelic blood-stage malaria viral-vectored vaccines and controlled malaria infection.
IL-7 licenses activation of human liver intrasinusoidal mucosal-associated invariant T cells.
Ulcerative colitis impairs the acylethanolamide-based anti-inflammatory system reversal by 5-aminosalicylic acid and glucocorticoids.
Decreased NK cells in patients with head and neck cancer determined in archival DNA.
Epigenetic biomarkers of T-cells in human glioma.
Cytomegalovirus-specific T-cell reactivity in biliary atresia at the time of diagnosis is associated with deficits in regulatory T cells.
Evidence for local dendritic cell activation in pulmonary sarcoidosis.
Effects of NOD-like receptors in human B lymphocytes and crosstalk between NOD1/NOD2 and Toll-like receptors.
Monocyte surface expression of Fcgamma receptor RI (CD64), a biomarker reflecting type-I interferon levels in systemic lupus erythematosus.
Accumulation of natural killer T cells in progressive nonalcoholic fatty liver disease.
Binding and uptake of H-ferritin are mediated by human transferrin receptor-1.
T regulatory cell levels decrease in people infected with Schistosoma mansoni on effective treatment.
Mycobacterium tuberculosis-induced gamma interferon production by natural killer cells requires cross talk with antigen-presenting cells involving Toll-like receptors 2 and 4 and the mannose receptor in tuberculous pleurisy.
Expansion of regulatory T cells in patients with Langerhans cell histiocytosis.
Leon G, Hernandez Santana YE, Irwin N, Giannoudaki E, O'Neill S, Csizmadia I, Gogarty M, Lee TJ, Ruane D, Long A, Fallon PG, Hussey S, Walsh PT
Mucosal immunology 2022 Mar;15(3):491-503
Mucosal immunology 2022 Mar;15(3):491-503
Aberrant gut-microbiota-immune-brain axis development in premature neonates with brain damage.
Seki D, Mayer M, Hausmann B, Pjevac P, Giordano V, Goeral K, Unterasinger L, Klebermaß-Schrehof K, De Paepe K, Van de Wiele T, Spittler A, Kasprian G, Warth B, Berger A, Berry D, Wisgrill L
Cell host & microbe 2021 Oct 13;29(10):1558-1572.e6
Cell host & microbe 2021 Oct 13;29(10):1558-1572.e6
Single-cell RNA sequencing reveals ex vivo signatures of SARS-CoV-2-reactive T cells through 'reverse phenotyping'.
Fischer DS, Ansari M, Wagner KI, Jarosch S, Huang Y, Mayr CH, Strunz M, Lang NJ, D'Ippolito E, Hammel M, Mateyka L, Weber S, Wolff LS, Witter K, Fernandez IE, Leuschner G, Milger K, Frankenberger M, Nowak L, Heinig-Menhard K, Koch I, Stoleriu MG, Hilgendorff A, Behr J, Pichlmair A, Schubert B, Theis FJ, Busch DH, Schiller HB, Schober K
Nature communications 2021 Jul 26;12(1):4515
Nature communications 2021 Jul 26;12(1):4515
Dendritic Cell Maturation Regulates TSPAN7 Function in HIV-1 Transfer to CD4(+) T Lymphocytes.
Perot BP, García-Paredes V, Luka M, Ménager MM
Frontiers in cellular and infection microbiology 2020;10:70
Frontiers in cellular and infection microbiology 2020;10:70
Single-cell transcriptomics reveals regulators underlying immune cell diversity and immune subtypes associated with prognosis in nasopharyngeal carcinoma.
Chen YP, Yin JH, Li WF, Li HJ, Chen DP, Zhang CJ, Lv JW, Wang YQ, Li XM, Li JY, Zhang PP, Li YQ, He QM, Yang XJ, Lei Y, Tang LL, Zhou GQ, Mao YP, Wei C, Xiong KX, Zhang HB, Zhu SD, Hou Y, Sun Y, Dean M, Amit I, Wu K, Kuang DM, Li GB, Liu N, Ma J
Cell research 2020 Nov;30(11):1024-1042
Cell research 2020 Nov;30(11):1024-1042
Aging affects responsiveness of peripheral blood mononuclear cells to immunosuppression of periodontal ligament stem cells.
Li X, Zhang B, Wang H, Zhao X, Zhang Z, Ding G, Wei F
The Journal of international medical research 2020 Jul;48(7):300060520930853
The Journal of international medical research 2020 Jul;48(7):300060520930853
Involvement of natural killer cells in the pathogenesis of endometriosis in patients with pelvic pain.
He J, Xu Y, Yi M, Gu C, Zhu Y, Hu G
The Journal of international medical research 2020 Jul;48(7):300060519871407
The Journal of international medical research 2020 Jul;48(7):300060519871407
Th2-like T Follicular Helper Cells Promote Functional Antibody Production during Plasmodium falciparum Infection.
Chan JA, Loughland JR, de Labastida Rivera F, SheelaNair A, Andrew DW, Dooley NL, Wines BD, Amante FH, Webb L, Hogarth PM, McCarthy JS, Beeson JG, Engwerda CR, Boyle MJ
Cell reports. Medicine 2020 Dec 22;1(9):100157
Cell reports. Medicine 2020 Dec 22;1(9):100157
Skewed T cell responses to Epstein-Barr virus in long-term asymptomatic kidney transplant recipients.
Nakid-Cordero C, Arzouk N, Gauthier N, Tarantino N, Larsen M, Choquet S, Burrel S, Autran B, Vieillard V, Guihot A
PloS one 2019;14(10):e0224211
PloS one 2019;14(10):e0224211
Inflammation induced by incomplete radiofrequency ablation accelerates tumor progression and hinders PD-1 immunotherapy.
Shi L, Wang J, Ding N, Zhang Y, Zhu Y, Dong S, Wang X, Peng C, Zhou C, Zhou L, Li X, Shi H, Wu W, Long X, Wu C, Liao W
Nature communications 2019 Nov 28;10(1):5421
Nature communications 2019 Nov 28;10(1):5421
Loss of ARPC1B impairs cytotoxic T lymphocyte maintenance and cytolytic activity.
Randzavola LO, Strege K, Juzans M, Asano Y, Stinchcombe JC, Gawden-Bone CM, Seaman MN, Kuijpers TW, Griffiths GM
The Journal of clinical investigation 2019 Dec 2;129(12):5600-5614
The Journal of clinical investigation 2019 Dec 2;129(12):5600-5614
Biallelic mutations in DNA ligase 1 underlie a spectrum of immune deficiencies.
Maffucci P, Chavez J, Jurkiw TJ, O'Brien PJ, Abbott JK, Reynolds PR, Worth A, Notarangelo LD, Felgentreff K, Cortes P, Boisson B, Radigan L, Cobat A, Dinakar C, Ehlayel M, Ben-Omran T, Gelfand EW, Casanova JL, Cunningham-Rundles C
The Journal of clinical investigation 2018 Dec 3;128(12):5489-5504
The Journal of clinical investigation 2018 Dec 3;128(12):5489-5504
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
Follicular Regulatory T Cells Are Highly Permissive to R5-Tropic HIV-1.
Miller SM, Miles B, Guo K, Folkvord J, Meditz AL, McCarter MD, Levy DN, MaWhinney S, Santiago ML, Connick E
Journal of virology 2017 Sep 1;91(17)
Journal of virology 2017 Sep 1;91(17)
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
The 4E-BP-eIF4E axis promotes rapamycin-sensitive growth and proliferation in lymphocytes.
So L, Lee J, Palafox M, Mallya S, Woxland CG, Arguello M, Truitt ML, Sonenberg N, Ruggero D, Fruman DA
Science signaling 2016 May 31;9(430):ra57
Science signaling 2016 May 31;9(430):ra57
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
Dendritic cell-activated cytokine-induced killer cell-mediated immunotherapy is safe and effective for cancer patients >65 years old.
Liu Y, Liu H, Liu H, He P, Li J, Liu X, Chen L, Wang M, Xi J, Wang H, Zhang H, Zhu Y, Zhu W, Ning J, Guo C, Sun C, Zhang M
Oncology letters 2016 Dec;12(6):5205-5210
Oncology letters 2016 Dec;12(6):5205-5210
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
Characterization of the in vivo immune network of IDO, tryptophan metabolism, PD-L1, and CTLA-4 in circulating immune cells in melanoma.
Chevolet I, Speeckaert R, Schreuer M, Neyns B, Krysko O, Bachert C, Hennart B, Allorge D, van Geel N, Van Gele M, Brochez L
Oncoimmunology 2015 Mar;4(3):e982382
Oncoimmunology 2015 Mar;4(3):e982382
Clinical significance of plasmacytoid dendritic cells and myeloid-derived suppressor cells in melanoma.
Chevolet I, Speeckaert R, Schreuer M, Neyns B, Krysko O, Bachert C, Van Gele M, van Geel N, Brochez L
Journal of translational medicine 2015 Jan 16;13:9
Journal of translational medicine 2015 Jan 16;13:9
Serum macrophage-derived chemokine/CCL22 levels are associated with glioma risk, CD4 T cell lymphopenia and survival time.
Zhou M, Bracci PM, McCoy LS, Hsuang G, Wiemels JL, Rice T, Zheng S, Kelsey KT, Wrensch MR, Wiencke JK
International journal of cancer 2015 Aug 15;137(4):826-36
International journal of cancer 2015 Aug 15;137(4):826-36
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
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
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
HER2 as a promising target for cytotoxicity T cells in human melanoma therapy.
Ma J, Han H, Liu D, Li W, Feng H, Xue X, Wu X, Niu G, Zhang G, Zhao Y, Liu C, Tao H, Gao B
PloS one 2013;8(8):e73261
PloS one 2013;8(8):e73261
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
Evaluation of clinical and immunological responses: a 2-year follow-up study in children with allergic rhinitis due to house dust mite.
Moed H, Gerth van Wijk R, Hendriks RW, van der Wouden JC
Mediators of inflammation 2013;2013:345217
Mediators of inflammation 2013;2013:345217
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
Inhibition of CXCR3-mediated chemotaxis by the human chemokine receptor-like protein CCX-CKR.
Vinet J, van Zwam M, Dijkstra IM, Brouwer N, van Weering HR, Watts A, Meijer M, Fokkens MR, Kannan V, Verzijl D, Vischer HF, Smit MJ, Leurs R, Biber K, Boddeke HW
British journal of pharmacology 2013 Mar;168(6):1375-87
British journal of pharmacology 2013 Mar;168(6):1375-87
Association of a NOD2 gene polymorphism and T-helper 17 cells with presumed ocular toxoplasmosis.
Dutra MS, Béla SR, Peixoto-Rangel AL, Fakiola M, Cruz AG, Gazzinelli A, Quites HF, Bahia-Oliveira LM, Peixe RG, Campos WR, Higino-Rocha AC, Miller NE, Blackwell JM, Antonelli LR, Gazzinelli RT
The Journal of infectious diseases 2013 Jan 1;207(1):152-63
The Journal of infectious diseases 2013 Jan 1;207(1):152-63
Assessment of immune interference, antagonism, and diversion following human immunization with biallelic blood-stage malaria viral-vectored vaccines and controlled malaria infection.
Elias SC, Collins KA, Halstead FD, Choudhary P, Bliss CM, Ewer KJ, Sheehy SH, Duncan CJ, Biswas S, Hill AV, Draper SJ
Journal of immunology (Baltimore, Md. : 1950) 2013 Feb 1;190(3):1135-47
Journal of immunology (Baltimore, Md. : 1950) 2013 Feb 1;190(3):1135-47
IL-7 licenses activation of human liver intrasinusoidal mucosal-associated invariant T cells.
Tang XZ, Jo J, Tan AT, Sandalova E, Chia A, Tan KC, Lee KH, Gehring AJ, De Libero G, Bertoletti A
Journal of immunology (Baltimore, Md. : 1950) 2013 Apr 1;190(7):3142-52
Journal of immunology (Baltimore, Md. : 1950) 2013 Apr 1;190(7):3142-52
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
Decreased NK cells in patients with head and neck cancer determined in archival DNA.
Accomando WP, Wiencke JK, Houseman EA, Butler RA, Zheng S, Nelson HH, Kelsey KT
Clinical cancer research : an official journal of the American Association for Cancer Research 2012 Nov 15;18(22):6147-54
Clinical cancer research : an official journal of the American Association for Cancer Research 2012 Nov 15;18(22):6147-54
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
Cytomegalovirus-specific T-cell reactivity in biliary atresia at the time of diagnosis is associated with deficits in regulatory T cells.
Brindley SM, Lanham AM, Karrer FM, Tucker RM, Fontenot AP, Mack CL
Hepatology (Baltimore, Md.) 2012 Apr;55(4):1130-8
Hepatology (Baltimore, Md.) 2012 Apr;55(4):1130-8
Evidence for local dendritic cell activation in pulmonary sarcoidosis.
Ten Berge B, Kleinjan A, Muskens F, Hammad H, Hoogsteden HC, Hendriks RW, Lambrecht BN, Van den Blink B
Respiratory research 2012 Apr 18;13(1):33
Respiratory research 2012 Apr 18;13(1):33
Effects of NOD-like receptors in human B lymphocytes and crosstalk between NOD1/NOD2 and Toll-like receptors.
Petterson T, Jendholm J, Månsson A, Bjartell A, Riesbeck K, Cardell LO
Journal of leukocyte biology 2011 Feb;89(2):177-87
Journal of leukocyte biology 2011 Feb;89(2):177-87
Monocyte surface expression of Fcgamma receptor RI (CD64), a biomarker reflecting type-I interferon levels in systemic lupus erythematosus.
Li Y, Lee PY, Kellner ES, Paulus M, Switanek J, Xu Y, Zhuang H, Sobel ES, Segal MS, Satoh M, Reeves WH
Arthritis research & therapy 2010;12(3):R90
Arthritis research & therapy 2010;12(3):R90
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
Binding and uptake of H-ferritin are mediated by human transferrin receptor-1.
Li L, Fang CJ, Ryan JC, Niemi EC, Lebrón JA, Björkman PJ, Arase H, Torti FM, Torti SV, Nakamura MC, Seaman WE
Proceedings of the National Academy of Sciences of the United States of America 2010 Feb 23;107(8):3505-10
Proceedings of the National Academy of Sciences of the United States of America 2010 Feb 23;107(8):3505-10
T regulatory cell levels decrease in people infected with Schistosoma mansoni on effective treatment.
Watanabe K, Mwinzi PN, Black CL, Muok EM, Karanja DM, Secor WE, Colley DG
The American journal of tropical medicine and hygiene 2007 Oct;77(4):676-82
The American journal of tropical medicine and hygiene 2007 Oct;77(4):676-82
Mycobacterium tuberculosis-induced gamma interferon production by natural killer cells requires cross talk with antigen-presenting cells involving Toll-like receptors 2 and 4 and the mannose receptor in tuberculous pleurisy.
Schierloh P, Yokobori N, Alemán M, Landoni V, Geffner L, Musella RM, Castagnino J, Baldini M, Abbate E, de la Barrera SS, Sasiain MC
Infection and immunity 2007 Nov;75(11):5325-37
Infection and immunity 2007 Nov;75(11):5325-37
Expansion of regulatory T cells in patients with Langerhans cell histiocytosis.
Senechal B, Elain G, Jeziorski E, Grondin V, Patey-Mariaud de Serre N, Jaubert F, Beldjord K, Lellouch A, Glorion C, Zerah M, Mary P, Barkaoui M, Emile JF, Boccon-Gibod L, Josset P, Debré M, Fischer A, Donadieu J, Geissmann F
PLoS medicine 2007 Aug;4(8):e253
PLoS medicine 2007 Aug;4(8):e253
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Staining of human peripheral blood mononuclear cells with CD45 Pacific Blue, CD19 APC and CD3 FITC. As expected based on known relative expression patterns, CD3 clone UCHT1 stains a subset of lymphocytes (pink), but not monocytes (orange) and granulocytes (blue).
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Supportive validation
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- Figure 6 Representative high-magnification photomicrographs showing double immunofluorescence for NAAA, CD19, CD3 and CD14 in order to characterize the immune cells in the mucosa infiltrate of UC patients. NAAA immunofluorescence was observed in CD19+ B lymphocytes (A-C), CD3+ T lymphocytes (D-F) and CD14+ macrophages (G-I).
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- Figure 9 Representative high-magnification photomicrographs showing double immunofluorescence for FAAH, NAPE-PLD, CD38 and CD3 in order to characterize the immune cells in the mucosa infiltrate of UC patients. Nearly all FAAH immunofluorescent cells are plasma cell-specific CD38 (A-C). NAPE-PLD immunofluorescence was observed in both CD38+ plasma cells (D-F) and CD3+ T lymphocytes (G-I).
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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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- Fig. 4 T/NK cell clusters in NPC. a t-SNE plot showing 10 clusters of 17,263 T/NK cells (indicated by colors). b t-SNE plot, color coding for the expression of the marker genes (gray to red) for the indicated cell subtypes. c Average expression of selected T cell function-associated genes of naive markers, inhibitory receptors, cytokines and effector molecules, co-stimulatory molecules, and Treg markers in each cell cluster. d Potential developmental trajectory of CD4 + T cells ( n = 5694) inferred by analysis with Monocle 2. Arrows show the increasing directions of certain CD4 + T cell properties annotated with the signatures shown in e . e Traceplots of (left) CD4 + T cell activation signature along activation component and (right) terminal differentiation signature along terminal differentiation component for the CD4 + T cells. Cells are projected along the component, with the blue line indicating the moving average of the expression of signatures (a sliding window of length equal to 5% of the total number of CD4 + T cells was used), and the shaded area displaying SEM. Signatures used are presented in Supplementary information, Table S9 . f Potential developmental trajectory of CD8 + T cells ( n = 6975) inferred by analysis with Monocle 2. Arrows show the increasing directions of certain CD8 + T cell properties annotated with the signatures shown in g . g Traceplots (as in e ) of (left) CD8 + T cell activation signature along activation component and (right) terminal diffe
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- Fig. 6 The dense network and multiple regulatory immune responses in the TME of NPC. a Capacity for intercellular communication between malignant cells and immune cells. Each line color indicates the ligands expressed by the cell population represented in the same color (labeled). The lines connect to the cell types that express the cognate receptors. The line thickness is proportional to the number of ligands when cognate receptors are present in the recipient cell type. The loops indicate autocrine circuits. The map quantifies potential communication but does not account for the anatomical locations or boundaries of the cell types. b Detailed view of the ligands expressed by each major cell type and the cells expressing the cognate receptors primed to receive the signal. Numbers indicate the quantity of ligand-receptor pairs for each intercellular link. c - f Overview of selected ligand-receptor interactions of tumor cells ( c ), dysfunctional CD8 + T cells ( d ), macrophages ( e ), and the three types of DCs ( f , DC1, DC2, and DC3). P values are indicated by circle size, with the scale to the right (permutation test). The means of the average expression levels of interacting molecule 1 in cluster 1 and interacting molecule 2 in cluster 2 are indicated by color. Assays were carried out at the mRNA level but were used to extrapolate protein interactions. CD4T conv , conventional CD4 + T cell; CD8T, CD8 + T cell; CD8T dys , dysfunctional CD8 + T cell; DC, dendritic cell; GCB
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- Figure 2 T cell ontogenesis in extremely premature infants with and without severe brain injury (A) Principal coordinates analysis (PCoA)--biplot of sequestered cytokine and chemokine composition. Silhouette scoring identified 3 main cyto-Clusters (as indicated by different symbols). The shade of the symbols (gray to black) is determined by days post-delivery--the older the infant the darker the symbol. The size of the symbols is determined by the Kidokoro Score as assessed at term-equivalent age via cMRI--the higher the score the larger the symbol. Significantly correlated (p < 0.05) cytokines/chemokines are plotted as arrows. In addition, box plots next to PCoA show the range of combined values for burst suppression and variance of cranial oxygenation (BS/100 x VAR-cSO 2 ), as well as the range of Kidokoro Scores in the respective cyto-clusters (cyto-cluster 1, CC1; cyto-cluster 2, CC2; cyto-cluster 3, CC3). (B) Blood cytokine/chemokine concentrations in infants with (red) and without (blue) severe BI (3 days post-delivery, d3; 7 days post-delivery, d7; 28 days post-delivery, d28; 32 weeks of gestational age, w32; term-equivalent age = term). (C) Representative images illustrating the gating strategy for FACS analysis. Differently colored gates mark gating for respective cell populations. Magenta, untargeted; blue, T helper cells; orange, cytotoxic T cells; green, T regulatory cells; pink, gammadelta T cells. For box plots in (D) and (F-J), darker shade represents data from
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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
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- Figure 7 Absence of ARPC1B alters surface expression of CD8 and GLUT1 in hCTLs. (A-D) HD and ARPC1B-deficient patient hCTLs were fixed in PFA for 20 minutes, permeabilized, and stained with an antibody against CD8 alone (green) ( A and B ) or in combination with anti-GLUT1 (red) and anti-EEA1 (white) antibodies ( C and D ). Images are 3D reconstructions of Z -stack. Scale bars: 4 mum. ( E ) Measurement of the mean intensity of GLUT1 expressed in AU and the degree of colocalization with EEA1 expressed as PCC (see Methods) in HD and ARPC1B-deficient patient hCTLs based on images as sampled in C and D . HD, n = 41 cells; ARPC1B-deficient patient, n = 38 cells. P < 0.005 (unpaired t test). ** P < 0.0013; *** P < 0.0002. ( F ) Flow cytometry analysis of the proliferation capacity of HD and ARPC1B-deficient patient hCTLs (gated on live CD8 + cells) in the absence (blue) or presence (red) of plate-bound anti-CD3 stimulation (1 mug/mL). All data are representative of 3 independent experiments.
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- Figure 5 CD169, as an HIV-1 receptor, mostly impacts transfer from mature MDDCs rather than immature MDDCs. (A) Flow cytometry plots showing CD86, DC-SIGN, and CD169 expression levels on MDDCs (pre-gated on SSC FSC, living cells, CD3 - cells and singlets). Panels show the expression of these proteins in iMDDCs (left panel) and MDDCs with LPS pretreatment at 100 ng/ml for 48 or 24 h before co-culture (middle and right panels, respectively). (B) Percentage of variation of HIV-1 transfer when using iMDDCs or LPS-treated MDDCs (100 ng/ml LPS for different lengths of time) incubated with a blocking antibody against CD169 as compared to an isotype control for each condition. Results are displayed for 4 different blood donors with the mean +- SD of technical triplicates. (C) Percent of variation in HIV-1 transfer to assess the impact of blocking CD169 and TSPAN7 knockdown as compared to scramble shRNA on MDDCs matured with LPS for 48 h treated by an isotype control. Mean +- SD of seven different blood donors in 4 experiments. (B,C) NS, not significant. ** p < 0.01; *** p < 0.001. (D) Confocal microscopy images of iMDDCs (left panel) and mature MDDCs (mMDDCs) right panel, to assess the degree of colocalization between CD169 (magenta) and incoming X4-HIV-1-Gag-iGFP (green). Actin filaments and nuclei were stained with phalloidin (red) and DAPI (blue). Four hundred nanometers of Z-stacks were taken 40 h after the start of the co-culture with CD4 + T cells and X4-HIV-1-Gag-iGFP. The pic
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- Fig. 5 Infiltrating myeloid cells inhibit T cell functionality in residual tumors. a Flow cytometric analysis of CD11b + , CD3 + and the ratio of CD3 + /CD11b + in the untreated and iRFA-treated CT26 and MC38 tumors on day 3 and day 9 ( n = 5). b Flow cytometric analysis of CD8 + and CD4 + FoxP3 + cells and the ratio of CD8 + /CD4 + FoxP3 + in the untreated and iRFA-treated CT26 and MC38 tumors on day 3 and day 9 ( n = 5). c Flow cytometric analysis and quantification of granzyme B, IFN-gammaand PD-1 expression in CD8 + cells in the untreated and iRFA-treated CT26 and MC38 tumors on day 3 and day 9 ( n = 5). d In vitro suppressive activity of tumor-infiltrating CD11b + cells purified from spleen or CT26 residual tumors on day 3 after iRFA. Representative histograms of CD8 + T cell proliferation at a ratio of 1:1 CD8 + to CD11b + T cells and percent CD8 + T cell proliferation ( n = 5). e - g CD11b + cells isolated from the residual tumor were mixed with CT26 or MC38 tumor cells and transferred into recipient mice. e Adoptive transfer method. f Growth curve of tumor (one-sided ANOVA test, *** P < 0.001, n = 5). g Flow cytometric analysis and quantification of CD3 + CD8 + cells (gate on single live cells) and Granzyme B expression and IFN-gamma on CD8 + cells ( n = 5). Data represent cumulative results from 1/2 independent experiments with 5 mice/group. The data are represented as mean +- SEM. Statistical differences between pairs of groups
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- Fig. 8 CCL2/CCR2 blockade inhibits tumor progression and overcomes resistance to anti-PD-1 therapy. a - f iRFA treatment was performed in CT26 and MC38 colon cancer models as shown in Fig. 2a . Anti-PD-1 mAb (200 mug, clone: J43) was administered through intraperitoneal injection to mice every 3 days for a total of four times. The CCR2 antagonist (CCR2a) (RS504393, Tocris) was given subcutaneously at a dose of 5 mg/kg twice per day for 9 days. a Growth curve of the CT26 and MC38 residual tumor (one-sided ANOVA test, n = 8). b The weight of the residual CT26 and MC38 tumor examined on day 14 after iRFA by dissection of the mice ( n = 6). c The number of metastases examined on day 14 after iRFA by dissection the mice ( n = 6). d Kaplan-Meier survival curves are shown, and the log-rank test was performed ( n = 8). e Flow cytometric analysis and quantification of CD3 + and CD8 + infiltration (gate on single live cells) in residual CT26 tumors. f Granzyme B and IFNgamma expression on CD8 + cells in residual CT26 tumors. (gate on CD8 + cells) ( n = 5). g , h iRFA treatment was performed in mice bearing wild type and CCL2 -/- CT26 or MC38 tumor. g Growth curve of the CT26 and MC38 residual tumor (one-sided ANOVA test, n = 5). h Kaplan-Meier survival curves are shown, and the log-rank test was performed ( n = 8). Data represent results from 1/2 independent experiments. The data are represented as mean +- SEM. Statistical differences between pairs o
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- Figure 1 Proliferating Cells in LCH Granuloma are Mostly Endothelial Cells, Fibroblasts, and T Cells Paraffin-embedded and frozen sections were stained with antibodies against Ki-67 (which label proliferating cells), CD1a (LCs), CD3 (T cells), CD20 (B cells), CD68, CD31, and CD34 (endothelial cells). (A) Double immunostaining of paraffin-embedded section from LCH eosinophilic granulomas with anti-Ki-67 Ab, (brown nuclear staining) and with anti-CD1a Ab (upper images, blue staining) or anti-CD3 Ab (lower images, blue staining). Open arrowheads indicate double-stained cells, black arrowheads indicate Ki-67 + cells with an endothelial morphology. (B) Histogram represents percentage of CD1a + cells and of CD3 + cells labeled with Ki-67 ( n = 15). (C) Histogram represents percentage of proliferating cells (Ki-67 + ) that express CD1a, CD3, CD20, or CD68 ( n = 15). (D) Histogram represents percentage of proliferating cells (Ki-67 + ) that are endothelial cells, interstitial cells (fibroblasts), and other types based on morphological examination. (E) Immunolabeling of blood vessels on paraffin-embedded section from LCH eosinophilic granulomas with CD34 (left) and CD31 (right) antibodies. (F) Proliferating Ki-67 + cells (brown nuclear staining) with a fibroblast-cell morphology in an eosinophilic granuloma.
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