KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-human/mouse BCL6 PerCP-Cy5.5 | Biolegend | RRID:AB_2566191 |
| Anti-human CD279 PE | Biolegend | RRID:AB_940483 |
| Anti-human CD3 BV421 | Biolegend | RRID:AB_2565849 |
| Anti-human CD4 BV510 | Biolegend | RRID:AB_2561377 |
| Anti-human CD8 APC | Biolegend | RRID:AB_2075390 |
| Anti-human CD185 BV711 | Biolegend | RRID:AB_2629525 |
| Anti-human CD278 BV650 | Biolegend | RRID:AB_2749928 |
| Anti-human CD45RA APC-Cy7 | Biolegend | RRID:AB_10708419 |
| Anti-human CD45RO PE-Cy7 | Biolegend | RRID:AB_11203903 |
| Anti-human FOXP3 AF488 | Biolegend | RRID:AB_430882 |
| Anti-human CD25 BV785 | Biolegend | RRID:AB_11219197 |
| Anti-human CD45 AF488 | Biolegend | RRID:AB_493033 |
| Anti-human CD20 BV650 | Biolegend | RRID:AB_11218609 |
| Anti-human CD27 BV785 | Biolegend | RRID:AB_2562674 |
| Anti-human CD38 APC-Cy7 | Biolegend | RRID:AB_2562576 |
| Anti-human CD138 BV421 | Biolegend | RRID:AB_2562659 |
| Anti-human IgG Fc APC | Biolegend | RRID:AB_11150590 |
| Anti-human IgM PE-Cy7 | Biolegend | RRID:AB_2566484 |
| Anti-human IgD BV510 | Biolegend | RRID:AB_2561386 |
| Anti-human CD21 PerCP-Cy5.5 | Biolegend | RRID:AB_2561543 |
| Anti-human CD45 BV785 | Biolegend | RRID:AB_2715888 |
| Anti-human CD3 BV605 | Biolegend | RRID:AB_11126166 |
| Anti-human CD19 BV711 | Biolegend | RRID:AB_2562062 |
| Anti-human CD56 AF488 | Biolegend | RRID:AB_2564093 |
| Anti-human CD11b PerCP-Cy5.5 | Biolegend | RRID:AB_10900072 |
| Anti-human CD68 APC-Cy7 | Biolegend | RRID:AB_2571964 |
| Anti-human CD14 PE-Cy7 | Biolegend | RRID:AB_830690 |
| Anti-human CD1c APC | Biolegend | RRID:AB_10718511 |
| Anti-human HLA-DR BV421 | Biolegend | RRID:AB_10897449 |
| Anti-human CD16 BV510 | Biolegend | RRID:AB_2562085 |
| Anti-human CD56 BV421 | Biolegend | RRID:AB_10900228 |
| Anti-human CD27 AF488 | Biolegend | RRID:AB_2565545 |
| Anti-human CD107a AF647 | Biolegend | RRID:AB_1227506 |
| Anti-human CD107a PE-Cy7 | Biolegend | RRID:AB_11147955 |
| Anti-human CD3 PE | Biolegend | RRID:AB_314044 |
| Anti-human CD19 APC | Biolegend | RRID:AB_314242 |
| Anti-human CD4 BV711 | Biolegend | RRID:AB_11219404 |
| Anti-human CXCR5 BV785 | Biolegend | RRID:AB_2629527 |
| Anti-human CXCR5 AF488 | Biolegend | RRID:AB_2561894 |
| Anti-human CD279 BB700 | BD Biosciences | RRID:AB_2744348 |
| Anti-human CD19 BV421 | Biolegend | RRID:AB_10897802 |
| Anti-human CD3 PE-Cy7 | Biolegend | RRID:AB_439781 |
| Anti-human CXCR5 BB700 | BD Biosciences | RRID:AB_2739739 |
| Anti-human CD127 PE-Cy7 | Biolegend | RRID:AB_10899414 |
| Anti-human PD-1 APC | Biolegend | RRID:AB_940475 |
| Anti-human FOXP3 eF450 | Thermo Fisher | RRID:AB_1834364 |
| Anti-human CD274 PE | Biolegend | RRID:AB_940368 |
| Anti-human CD274 APC | Biolegend | RRID:AB_2749927 |
| Anti-human PD-L1 | R&D Systems | RRID:AB_2073445 |
| Normal goat IgG control Antibody | R&D Systems | RRID:AB_354267 |
| Anti-human CD3 | Bio X Cell | RRID:AB_1107632 |
| Bacterial and Virus Strains | ||
| DH5a E. coli | NEB | Cat# C2987H |
| XL10 Gold E. coli | Agilent | Cat# 200315 |
| Stbl3 E. coli | Invitrogen | Cat# C737303 |
| StellarComp E. coli | Clontech | Cat# 636763 |
| VSV-G pseudotyped lentiviral vector | Viral Vector Core, Cincinnati Children’s Hospital Medical Center | https://www.cincinnatichildrens.org/research/cores/translational-core-laboratory/viral-vector-core/team |
| Biological Samples | ||
| Human tonsillar tissue | Division of Pediatric Otolaryngology-Head and Neck Surgery (Cincinnati Children’s Hospital Medical Center) | https://www.cincinnatichildrens.org/research/divisions/o/otolaryngology |
| Human cord blood | Cell Manipulations Laboratory & Cell Processing Core (Cincinnati Children’s Hospital Medical Center) | https://www.cincinnatichildrens.org/research/cores/translational-core-laboratory/cell-manipulations-laboratory https://www.cincinnatichildrens.org/research/cores/translational-core-laboratory/cell-processing-core |
| Leukocyte reduction filters (source of human PBMC) | University of Cincinnati Hoxworth Blood Center | https://hoxworth.org/ |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Collagenase D | Roche | Cat# 11088793001 |
| Recombinant Human IL-2 | Peprotech | Cat# 200–02 |
| DNase 1 | Millipore Sigma | Cat# 11284932001 |
| Ficoll-Paque PLUS | GE Healthcare | Cat# 17144002 |
| BD Cytofix Fixation Buffer | BD Biosciences | Cat# 554655 |
| BD Cytofix/Cytoperm Fixation and Permeabilization Solution | BD Biosciences | Cat# 554714 |
| Perm/Wash Buffer | BD Biosciences | Cat# 554723 |
| Ampicillin | GoldBio | Cat# A-301–25 |
| Kanamycin | GoldBio | Cat# K-120 |
| T4 DNA Ligase | New England BioLabs | Cat# M0202S |
| Human Fibronectin | Invitrogen | Cat# PHE0023 |
| Protamine Sulfate | Fisher | Cat# ICN10275205 |
| Blasticidin | Fisher | Cat# K515001 |
| Puromycin | GIBCO | Cat# A1113803 |
| rhPD-1-Fc | R&D Systems | Cat# 1086-PD-050 |
| IgG1-Fc | R&D Systems | Cat# 110-HG-100 |
| PMA | Sigma-Aldrich | Cat# P8139 |
| Ionomycin | Sigma-Aldrich | Cat# I9657–1MG |
| Zombie UV Dye | Biolegend | Cat# 423107 |
| SEB | EMD Millipore | Cat# 324798; CAS# 11100–45-1 |
| Calcein AM Solution | Fisher | Cat# C34852 |
| Propidium Iodide | Biolegend | Cat# 421301 |
| Chromium-51 | Perkin Elmer | Cat# NEZ030S002MC |
| CellTrace Violet | Invitrogen | Cat# C34557 |
| Pristane | Sigma-Aldrich | Cat# P9622–10X1ML |
| Bulsulfan | Sigma-Aldrich | Cat# B2635–25G |
| UltraComp eBeads Compensation Beads | ThermoFisher | Cat# 01–2222-42 |
| Critical Commercial Assays | ||
| DES Blasticidin Support Kit | Invitrogen | Cat# K515001 |
| DES-Inducible Kit with pCoBlast | Invitrogen | Cat# K512001 |
| GeneJET Gel Extraction kit | ThermoFisher | Cat# K0692 |
| QIAprep Mini, Midi, Maxi Kits | QIAGEN | Cat# 27106, 12943, 12362 |
| Rneasy Mini Kit | QIAGEN | Cat# 74104 |
| iScript cDNA Synthesis Kit | Bio-Rad | Cat# 170–8891 |
| PrimeTime Gene Expression Master Mix | IDT Technologies | Cat# 1055770 |
| TaqMan Gene Expression Assay | ThermoFisher | Cat# 4331182 |
| Easy Sep Human CD4+ T cell Isolation kit | StemCell Technologies | Cat# 17952 |
| Easy Sep Human B cell Isolation kit | StemCell Technologies | Cat# 17954 |
| Human IgG total ELISA Ready-SET-Go! Kit | eBioscience | Cat# 50–112-8669 |
| Experimental Models: Cell Lines | ||
| Drosophila S2 cell line | ThermoFisher | Cat# R69007 |
| HEK293T cell line | ATCC | RRID:CVCL_0063 |
| NK-92 cell line | ATCC | RRID:CVCL_2142 |
| Raji cell line | ATCC | RRID:CVCL_0511 |
| Experimental Models: Organisms/Strains | ||
| Mouse: NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ | Wunderlich et al., 201050 | RRID:IMSR_JAX:013062 |
| Oligonucleotides | ||
| Primer/probe sets for CAR expression qPCR | This paper | Table S1 |
| Sequencing primers PD-L1 CAR plasmid | This paper | Table S1 |
| Sequencing primers PD-1 pAc/V5-His plasmid | This paper | Table S1 |
| Sequencing primers PD-1 PB513 plasmid | This paper | Table S1 |
| Recombinant DNA | ||
| Human PDL1-CD28-CD3ζ chimeric antigen receptor | This paper | n/a |
| Human PD-1 protein sequence | Uniprot (a.a. 1–288) | n/a |
| pLVX-IRES-ZsGreen1 Vector | Clontech | Cat# 632187 |
| pUC57-Kan plasmid vector | GeneWiz | n/a |
| pAc5.1/V5-His A Plasmid Vector | Invitrogen | Cat# V411020 |
| PiggyBac Transposon, Cloning and Expression Vector | System Biosciences | Cat# PB513B-1 |
| Super PiggyBac Transposase Expression Vector | System Biosciences | Cat# PB210PA-1 |
| pcDNA3.1/His/lacZ | Invitrogen | Cat# V38520 |
| pAc5.1/V5-His A | Invitrogen | Cat# V411020 |
| Software and Algorithms | ||
| Sony SH800S Cell Sorter acquisition software | Sony Biotechnologies | https://www.sonybiotechnology.com/us/instruments/sh800s-cell-sorter/software/ |
| Snapgene | GSL Biotech | https://www.snapgene.com/ |
| Molecular Biology tool | Benchling | https://www.benchling.com/ |
| ‘‘Codon Optimization Tool’’ | IDT Technologies | https://www.idtdna.com/pages/tools |
| NIS-Elements Imaging Software | Nikon | https://www.microscope.healthcare.nikon.com/products/software |
| FACSDiva | BD Biosciences | https://www.bdbiosciences.com/en-us/instruments/research-instruments/research-software/flow-cytometry-acquisition/facsdiva-software |
| FlowJo v.10 Software | TreeStar Inc. | https://www.flowjo.com/ |
| Prism 7 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| Nonlinear Regression (curve fit) | GraphPad Prism | https://www.graphpad.com/guides/prism/7/curve-fitting/index.htm?REG_DR_stim_variable_2.htm. |