4 1bb 4b4 1

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4-1BB (4B4-1) is a cell surface receptor that belongs to the tumor necrosis factor receptor (TNFR) superfamily. It is primarily expressed on activated T cells and plays a role in the regulation of T cell responses.

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Lab products found in correlation

Rpa t8, by BD (1 mentions) Attune nxt flow cytometer, by Thermo Fisher Scientific (1 mentions) Cd16 3g8, by BD (1 mentions) Bcl 2, by BioLegend (1 mentions) Fortessa cytometer, by BD (1 mentions) Cd28 cd28.2, by BioLegend (1 mentions) Klrg1 sa231a2, by BioLegend (1 mentions)

2 protocols using 4 1bb 4b4 1

Multiparameter Phenotyping of CD8+ T Cells

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Cells were washed using phosphate-buffered saline (PBS) with 2% FBS, and were then stained with monoclonal antibodies (mAbs) specific for CD8 (RPA-T8, BD), CD45RA (HI100, BD), CD95 (DX-2, BD), CD62L (DREG-56, eBioscience, San Diego, CA), CD69 (FN50, BD), 4–1BB (4B4–1, Biolegend), ICOS (DX29, BD), CD27 (M-T271, BD), CD28 (CD28.2, BD), PD-1 (MIH4, BD), CD223 (LAG-3) (3DS223H, eBioscience), Tim-3 (F38–2E2, BD) and KLRG1 (13F12F2, eBioscience). MART-1- or gp100-specific CD8⁺ T cells were detected by MART-1 or gp100/HLA-A*0201 PE-labelled tetramers (MBL, Woburn, MA). Intracellular staining of Ki67 (B56, BD) was performed after fixation and permeabilization using a staining kit (eBioscience) according to the manufacturer’s instructions. Live Dead dyes (Invitrogen, Carlbad, CA) was used to assess viable cells. The cells in all experiments were analyzed with an Attune NXT flow cytometer (ThermoFisher Scientific, Waltham, MA) and analysed using FlowJo 10 software (Ashland, Oregon).

Ichikawa J., Yoshida T., Isser A.Y., Laino A.S., Vassallo M., Woods D.M., Kim S., Oelke M., Jones K., Schneck J.P, & Weber J.S. (2020). Rapid Expansion of Highly Functional Antigen-Specific T cells from Melanoma Patients by Nanoscale Artificial Antigen Presenting Cells. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(13), 3384-3396.

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Comprehensive Immune Cell Profiling

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Blood samples were collected at the indicated time points, PBMCs isolated through the research cell bank at FHCRC by standard Ficoll-Hypaque gradient, and viably cryopreserved. Cells were analyzed by flow cytometry after permeabilization, fixation and staining with fluorochrome-conjugated monoclonal antibodies to CD4 (SK3; Becton Dickinson), CD8 (3B5; Invitrogen), CD19 (H1B19; Becton Dickinson), CD16 (3G8; Becton Dickinson), CCR7 (G043H7; Biolegend), CD45RO (UCHL1; Becton Dickinson), CD28 (CD28.2; Biolegend), KLRG1 (SA231A2; Biolegend), CD27 (L128; Biolegend), CXCR3 (G025H7; Biolegend), CD127 (A019D5; Biolegend), PD1 (ED12.2H7; Biolegend), Lag3 (2DS223H; eBioscience), 4-1BB (4B4-1; Biolegend), BCL2 (100; Biolegend), CTLA-4 (L3D10) and the above described tetramers. Cells were analyzed on a Fortessa cytometer (Becton Dickinson) and data analysis performed with FlowJo. Staining, acquisition and analyses were performed on all samples in a batch on same day and negative controls included.

Paulson K.G., Voillet V., McAfee M.S., Hunter D.S., Wagener F.D., Perdicchio M., Valente W.J., Koelle S.J., Church C.D., Vandeven N., Thomas H., Colunga A.G., Iyer J.G., Yee C., Kulikauskas R., Koelle D.M., Pierce R.H., Bielas J.H., Greenberg P.D., Bhatia S., Gottardo R., Nghiem P, & Chapuis A.G. (2018). Acquired cancer resistance to combination immunotherapy from transcriptional loss of class I HLA. Nature Communications, 9, 3868.

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