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Anti cd3 clone ucht1

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Anti-CD3 (clone UCHT1) is a monoclonal antibody that specifically binds to the CD3 antigen on the surface of human T cells. It is commonly used in various laboratory applications for the identification and enumeration of T cells.

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Lab products found in correlation

Anti cd28 clone cd28, by BD (5 mentions) Lsrii flow cytometer, by BD (3 mentions) Foxp3 staining kit, by Thermo Fisher Scientific (2 mentions) Anti cd19 clone sj25c1, by BD (2 mentions) Anti ikaros clone r32 1149, by BD (2 mentions) Facscanto 2, by BD (2 mentions) Normal medium without glucose, by Thermo Fisher Scientific (2 mentions) Anti cd8 clone sk1, by BD (2 mentions) Anti cd27 clone l128, by BD (2 mentions) Anti cd4 clone l 200, by BD (2 mentions) Anti cd38 clone ls 198, by Beckman Coulter (2 mentions) Anti cd45ra clone abb11, by Beckman Coulter (2 mentions) Anti hla dr clone l243, by BioLegend (2 mentions) Facs lysing solution, by BD (2 mentions) Soluble anti cd28 clone cd28, by BD (2 mentions) Sodium pyruvate, by Lonza (2 mentions) Anti gpr15 clone sa302a10, by BioLegend (2 mentions) Sb203580, by Cell Signaling Technology (1 mentions) Pcr mycoplasma detection kit, by Applied Biological Materials (1 mentions) Rpmi 1640 medium, by Lonza (1 mentions)

Close spelling variants of this product

Anti-CD3 Pacific Blue (clone UCHT1, Anti-CD3 antibody (clone UCHT1, BUV395-conjugated anti-CD3 (clone UCHT1; Anti-human CD3 AF700 (clone UCHT1) Pacific Blue-conjugated anti-CD3 (clone UCHT1) Anti-CD3 mAb (Clone UCHT1, Anti-human CD3-Pacific Blue (PB) (clone UCHT1) Anti-CD3 FITC (clone: UCHT1; Anti-human CD3 PE-Cy7 (clone UCHT1, Mouse anti-human CD3 (clone UCHT1,

16 protocols using anti cd3 clone ucht1

1

Activation of T Cells by Antigen-Presenting Cells

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TRuC-T and CAR-T cells were co-cultured overnight with CD19+ Nalm6-LUC target cells or CD19− K562 target cells at 1:1 ratio in triplicates. Alternatively, CH7C17 cells were co-cultured with DapDR1-ICAM1 cells loaded with different amounts of the HA306–318 peptide. T cell activation markers were analyzed using anti-human CD25 (clone BC96) (eBioscience, San Diego, CA), anti-human CD69 (clone FN50) and anti-CD3 (clone UCHT1) (BD Biosciences, San Jose, CA).
Baeuerle P.A., Ding J., Patel E., Thorausch N., Horton H., Gierut J., Scarfo I., Choudhary R., Kiner O., Krishnamurthy J., Le B., Morath A., Baldeviano G.C., Quinn J., Tavares P., Wei Q., Weiler S., Maus M.V., Getts D., Schamel W.W, & Hofmeister R. (2019). Synthetic TRuC receptors engaging the complete T cell receptor for potent anti-tumor response. Nature Communications, 10, 2087.
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2

Analyzing Immune Cell Populations

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Cryopreserved PBMCs were first stained with fluorescently labeled anti-CD3 (clone UCHT1) and anti-CD19 (clone SJ25C1; both from BD Biosciences). Afterward, cells were fixed and permeabilized with the FoxP3 staining kit (eBioscience), according to the manufacturer’s instructions, and incubated with fluorescently labeled anti-IKAROS (clone R32-1149; BD Biosciences).E4 (link) Cells were acquired on a FACSCanto II flow cytometer (BD Biosciences). Data were analyzed with FlowJo software (version X; TreeStar).
Bogaert D.J., Kuehn H.S., Bonroy C., Calvo K.R., Dehoorne J., Vanlander A.V., De Bruyne M., Cytlak U., Bigley V., De Baets F., De Baere E., Rosenzweig S.D., Haerynck F, & Dullaers M. (2017). A novel IKAROS haploinsufficiency kindred with unexpectedly late and variable B-cell maturation defects. The Journal of allergy and clinical immunology, 141(1), 432-435.e7.
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3

Assessing T-cell Responses to Dendritic Cells

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In the MLR assay, monocyte-derived dendritic cells (MoDCs) with or without p38-MAPK inhibitor (SB203580; Cell Signaling, Danvers, MA, USA) treatment were cocultured with allogeneic CD4+ or CD8+ T cells for 5 days with concurrent stimulation of 1 μg/mL anti-CD3 (clone UCHT1; BD Biosciences). T-cell proliferation was characterized by flow cytometry as the percentage of cells exhibiting carboxyfluorescein succinimidyl ester (CFSE) fluorescence dilution.
Chen P., Urzua C.A., Knickelbein J.E., Kim J.S., Li Z., Hannes S., Kuo D., Chaigne-Delalande B., Armbrust K., Tucker W., Liu B., Agrón E., Sen H.N, & Nussenblatt R.B. (2016). Elevated CD1c+ Myeloid Dendritic Cell Proportions Associate With Clinical Activity and Predict Disease Reactivation in Noninfectious Uveitis. Investigative Ophthalmology & Visual Science, 57(4), 1765-1772.
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4

Tumor-Induced T Cell Metabolism Modulation

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Human peripheral blood T cell subsets (106/ml) were activated with anti-CD3 (clone UCHT1, 2.5 μg/ml) and anti-CD28 (clone CD28.2, 1.25 μg/ml) antibodies (BD Bioscience) for 3–5 days, as indicated in the figure legends. To obtain tumor medium, primary ovarian cancer cells were cultured for 3 days, and subsequently were frozen and thawed 5 times along with the culture medium. The cancer cell culture supernatant was obtained through centrifugation (20,000g, 1 h, 4°C), and used to culture T cells with or without glucose (2 mg/ml, Sigma Aldrich) supplementation for 16–24 h. To obtain normal medium containing different concentrations of glucose, normal medium without glucose (Invitrogen) was supplemented with 1 or 5 mg/ml glucose. Chemical inhibitors of metabolic pathways, Notch signaling or EZH2 were added as indicated in figure legends. Total numbers of viable cells at the endpoint of experiments were 1–2 × 106 cells.
Zhao E., Maj T., Kryczek I., Li W., Wu K., Zhao L., Wei S., Crespo J., Wan S., Vatan L., Szeliga W., Shao I., Wang Y., Liu Y., Varambally S., Chinnaiyan A.M., Welling T.H., Marquez V.E., Kotarski J., Wang H., Wang Z., Zhang Y., Liu R., Wang G, & Zou W. (2015). Cancer mediates effector T cell dysfunction by targeting microRNAs and EZH2 via glycolysis restriction. Nature immunology, 17(1), 95-103.
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5

Tumor-Induced T Cell Metabolism Modulation

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Human peripheral blood T cell subsets (106/ml) were activated with anti-CD3 (clone UCHT1, 2.5 μg/ml) and anti-CD28 (clone CD28.2, 1.25 μg/ml) antibodies (BD Bioscience) for 3–5 days, as indicated in the figure legends. To obtain tumor medium, primary ovarian cancer cells were cultured for 3 days, and subsequently were frozen and thawed 5 times along with the culture medium. The cancer cell culture supernatant was obtained through centrifugation (20,000g, 1 h, 4°C), and used to culture T cells with or without glucose (2 mg/ml, Sigma Aldrich) supplementation for 16–24 h. To obtain normal medium containing different concentrations of glucose, normal medium without glucose (Invitrogen) was supplemented with 1 or 5 mg/ml glucose. Chemical inhibitors of metabolic pathways, Notch signaling or EZH2 were added as indicated in figure legends. Total numbers of viable cells at the endpoint of experiments were 1–2 × 106 cells.
Zhao E., Maj T., Kryczek I., Li W., Wu K., Zhao L., Wei S., Crespo J., Wan S., Vatan L., Szeliga W., Shao I., Wang Y., Liu Y., Varambally S., Chinnaiyan A.M., Welling T.H., Marquez V.E., Kotarski J., Wang H., Wang Z., Zhang Y., Liu R., Wang G, & Zou W. (2015). Cancer mediates effector T cell dysfunction by targeting microRNAs and EZH2 via glycolysis restriction. Nature immunology, 17(1), 95-103.
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6

Whole Blood Flow Cytometry Assay

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A whole blood flow cytometry technique was employed as previously described [47 (link)]. Whole-blood samples (200ul) were stained at room temperature for 20 minutes. The following directly conjugated monoclonal antibodies (mAbs) were used: anti-CD3 (clone UCHT1, BD Biosciences), anti-CD4 (clone L-200, BD Biosciences), anti-CD8 (clone SK1, BD Biosciences), anti-CD27 (clone L128, BD Biosciences), anti-CD38 (clone LS-198, Beckman Coulter), anti-CD45RA (clone AbB11, Beckman Coulter) and anti-HLA-DR (clone L243, Biolegend). Red cells were lysed and fixed in FACS lysing solution (BD Biosciences) for 10 minutes in the dark, washed twice in FACS buffer (phosphate-buffered saline (PBS) containing 2% bovine serum albumin and 0.1% NaN3), and fixed with 1% formalin in PBS. Cells were acquired on an LSR-II flow cytometer (BD Biosciences).
Kalokhe A.S., Ibegbu C.C., Kaur S.P., Amara R.R., Kelley M.E., del Rio C, & Stephenson R. (2016). Intimate Partner Violence is Associated with Increased CD4+ T-Cell Activation Among HIV-Negative High-Risk Women. Pathogens & Immunity, 1(1), 193-213.
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7

Isolation and culture of cancer cell lines and T cells

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A20, JeKo-1, Mino, Val, Maver-1 and Lewis lung carcinoma (LLC) cells were obtained from American Type Culture Collection (ATCC, USA). Mycoplasma testing was performed regularly (every 2 months) using Mycoplasma PCR detection kit (ABM, Canada). The number of passages between thawing and use in the described experiments ranged between two and five. Cells were cultured in RPMI-1640 medium supplemented with 15% fetal bovine serum, 100 U/mL penicillin, 100 μg/mL streptomycin, 2 mM l-glutamine, 25 mM HEPES, 100 μM nonessential amino acids, and 1 mM sodium pyruvate (Lonza).
Total CD3+ T cells were isolated from PBMCs derived from CLL patients by positive selection kit (Invitrogen, USA) and cultured with 0.5 μg/mL plate-bound anti-CD3 (clone UCHT1) and 0.5 μg/mL soluble anti-CD28 (clone CD28.2; BD Biosciences).
Pevonedistat (TAK924) was provided by Takeda Development Center Americas, Inc.
Wang X., Chen C., Vuong D., Rodriguez-Rodriguez S., Lam V., Roleder C., Wang J.H., Kambhampati S., Berger A., Pennock N., Torka P., Hernandez-Ilizaliturri F., Siddiqi T., Wang L., Xia Z, & Danilov A.V. (2023). Pharmacologic targeting of Nedd8-activating enzyme reinvigorates T-cell responses in lymphoid neoplasia. Leukemia, 37(6), 1324-1335.
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8

Whole Blood Flow Cytometry Method

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A whole blood flow cytometry technique was employed as previously
described [47 (link)]. Whole-blood samples
(200ul) were stained at room temperature for 20 minutes. The following directly
conjugated monoclonal antibodies (mAbs) were used: anti-CD3 (clone UCHT1, BD
Biosciences), anti-CD4 (clone L-200, BD Biosciences), anti-CD8 (clone SK1, BD
Biosciences), anti-CD27 (clone L128, BD Biosciences), anti-CD38 (clone LS-198,
Beckman Coulter), anti-CD45RA (clone AbB11, Beckman Coulter) and anti-HLA-DR
(clone L243, Biolegend). Red cells were lysed and fixed in FACS lysing solution
(BD Biosciences) for 10 minutes in the dark, washed twice in FACS buffer
(phosphate-buffered saline (PBS) containing 2% bovine serum albumin and
0.1% NaN3), and fixed with 1% formalin in PBS. Cells
were acquired on an LSR-II flow cytometer (BD Biosciences).
Kalokhe A.S., Ibegbu C.C., Kaur S.P., Amara R.R., Kelley M.E., del Rio C, & Stephenson R. (2016). Intimate Partner Violence is Associated with Increased CD4+ T-Cell Activation Among HIV-Negative High-Risk Women. Pathogens & Immunity, 1(1), 193-213.
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9

Analyzing Immune Cell Populations

Cited 1 time
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Cryopreserved PBMCs were first stained with fluorescently labeled anti-CD3 (clone UCHT1) and anti-CD19 (clone SJ25C1; both from BD Biosciences). Afterward, cells were fixed and permeabilized with the FoxP3 staining kit (eBioscience), according to the manufacturer’s instructions, and incubated with fluorescently labeled anti-IKAROS (clone R32-1149; BD Biosciences).E4 (link) Cells were acquired on a FACSCanto II flow cytometer (BD Biosciences). Data were analyzed with FlowJo software (version X; TreeStar).
Bogaert D.J., Kuehn H.S., Bonroy C., Calvo K.R., Dehoorne J., Vanlander A.V., De Bruyne M., Cytlak U., Bigley V., De Baets F., De Baere E., Rosenzweig S.D., Haerynck F, & Dullaers M. (2017). A novel IKAROS haploinsufficiency kindred with unexpectedly late and variable B-cell maturation defects. The Journal of allergy and clinical immunology, 141(1), 432-435.e7.
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10

Suppressor T Cell Functional Assay

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AutoMACS-sorted CD4+CD25+: “suppressor” T cells were cocultured in triplicate wells with autologous CD4+CD25− “responders” (60,000 cells/well) in 96-well plates at suppressor/responder ratios of 1:0, 1:1, 0.5:1, 0.25:1, 0.125:1, and 0:1 for 3 days with media alone, 2 μg/ml phytohemagglutinin (PHA; Sigma Chemical Co.), or 0.04 μg/ml anti-CD3 (clone UCHT1; BD Pharmingen) with 2 μg/ml anti-CD28 (clone CD28.2; BD Pharmingen) before 16 h of [3H]thymidine (1 μCi/well) uptake as previously described (7 (link), 33 (link), 36 (link)). Proliferation was expressed as a stimulation index (SI): the mean cpm in stimulated wells divided by the mean cpm in unstimulated wells. T cell proliferation in each coculture was normalized by proliferation in CD4+CD25− T cells alone and compared to the calculated percentage of FoxP3+CD4 T cells in each coculture condition, based on %FoxP3+ cells in CD4+CD25− and CD4+CD25+ cell subsets determined by FACS.
Cho H., Kikuchi M., Li Y., Nakamoto N., Amorosa V.K., Valiga M.E, & Chang K.M. (2014). Induction of Multiple Immune Regulatory Pathways with Differential Impact in HCV/HIV Coinfection. Frontiers in Immunology, 5, 265.
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