KEY RESOURCE TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER | |||
|---|---|---|---|---|---|
| Antibodies | |||||
| Anti-human CD3e | BioLegend | Cat#317326; clone OKT3 | |||
| Anti-human CD28 | BioLegend | Cat#302943; clone CD28.2 | |||
| Alexa Fluor 647 Mouse Anti-Stat5 (pY694) | BD Biosciences | Cat#612599; 47/Stat5(pY694) | |||
| Mouse Anti-Stat5 (pY694), PE | BD Biosciences | Cat#612567; 47/Stat5(pY694) | |||
| Mouse Anti-Stat3 (pY705), Alexa Fluor 647 | BD Biosciences | Cat#557815; clone 4/P-STAT3 | |||
| Phospho-Stat1 (Tyr701) Rabbit mAb, Alexa Fluor 647 Conjugate | Cell Signaling Technology | Cat#8009S; clone 58D6 | |||
| Phospho-Stat1 (Tyr701) Rabbit mAb, Alexa Fluor 488 Conjugate | Cell Signaling Technology | Cat#9174S; clone 58D6 | |||
| Phospho-S6 Ribosomal Protein (Ser240/244) XP Rabbit mAb, Alexa Fluor 488 Conjugate | Cell Signaling Technology | Cat#5018S; clone D68F8 | |||
| HA-Tag Mouse mAb, Alexa Fluor 647 Conjugate | Cell Signaling Technology | Cat#3444S; clone 6E2 | |||
| Ultra-LEAF Purified anti-human CD337 (NKp30) Antibody | BioLegend | Cat#325224; clone P30–15 | |||
| Anti-human CD107a (LAMP-1) Antibody, FITC | Biolegend | Cat#328606; clone H4A3 | |||
| BD GolgiPlug Protein Transport Inhibitor (containing Brefeldin A) | BD Biosciences | Cat#555029 | |||
| GolgiStop Protein Transport Inhibitor (Containing Monensin) | BD Biosciences | Cat#554724 | |||
| Fixation/Permeabilization Solution Kit | BD Biosciences | Cat#554714 | |||
| PE-Cy7 Mouse Anti-Human TNF | BD Biosciences | Cat#557647; clone Mab11 | |||
| Perm buffer III | BD Biosciences | Cat#558050 | |||
| Mouse Anti-Human CD8, PE | BD Biosciences | Cat#555635; clone HIT8a | |||
| Mouse Anti-Human MIP-1β, PE | BD Biosciences | Cat#550078; clone D21–1351 | |||
| Mouse Anti-Human IFN-γ, Alexa Fluor 647 | BD Biosciences | Cat#557729; clone B27 | |||
| Anti-human CD122 (IL-2Rβ) Antibody, APC | Biolegend | Cat#339008; clone TU27 | |||
| Anti-human CD3 Antibody, APC | Biolegend | Cat#300439; clone UCHT1 | |||
| Anti-human CD45RA Antibody, APC | Biolegend | Cat#304150; clone HI100 | |||
| Anti-human IFN-γ Antibody, APC | Biolegend | Cat#506510; clone B27 | |||
| Anti-human CD8 Antibody, APC/Cyanine7 | Biolegend | Cat#344714; clone SK1 | |||
| Anti-human CD56 (NCAM) Antibody, Brilliant Violet 605 | Biolegend | Cat#362538; clone 5.1H11 | |||
| Anti-human CD197 (CCR7) Antibody, Brilliant Violet 605 | Biolegend | Cat#353224; clone G043H7 | |||
| Anti-human CD8a Antibody, Brilliant Violet 605 | Biolegend | Cat#344742; clone SK1 | |||
| Anti-human CD4 Antibody, Brilliant Violet 785 | Biolegend | Cat#300554; clone RPA-T4 | |||
| Anti-human CD3 Antibody, FITC | Biolegend | Cat#300452; clone UCHT1 | |||
| Anti-human CD4 Antibody, FITC | Biolegend | Cat#300538; clone RPA-T4 | |||
| FITC anti-human CD69 Antibody | Biolegend | Cat#310904; Clone FN50 | |||
| Anti-human CD3 Antibody , Pacific Blue | Biolegend | Cat#300431; clone UCHT1 | |||
| Anti-human CD4 Antibody, Pacific Blue | Biolegend | Cat#300521; clone RPA-T4 | |||
| Anti-human IL-2 Antibody, Pacific Blue | Biolegend | Cat#500307; clone MQ117H12 | |||
| Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Rabbit mAb, Alexa Fluor 647 Conjugate | Biolegend | Cat#13148s; clone 197G2 | |||
| Phospho-Akt (Ser473) XP Rabbit mAb, Alexa Fluor 647 Conjugate | Cell Signaling Technology | Cat#4075S; clone DE9 | |||
| Bacterial and Virus Strains | |||||
| Mix & Go Competent Cells - DH5α | Zymo Research | Cat#T3007 | |||
| Biological Samples | |||||
| Human peripheral mononuclear cells (PBMCs), isolated from leukocyte reduction shuttles | Stanford Blood Center | N/A | |||
| Chemicals, Peptides, and Recombinant Proteins | |||||
| SA sensor chip | Cytiva | Cat#BR-1005–31 | |||
| HBS-P | Cytiva | Cat#BR-1006–71 | |||
| EasySep Magnet | StemCell Technologies | Cat#18000 | |||
| Human CD8+ T cell Isolation Kit | Miltenyi | Cat#130–096-495 | |||
| LS magnetic selection column | Miltenyi | Cat#130–042-401 | |||
| EasySep Human Naïve Pan T Cell Isolation Kit | StemCell Technologies | Cat#17961 | |||
| ExpiFectamine 293 Transfection Kit | Gibco | Cat#A14525 | |||
| Sapphire Baculovirus DNA | Allele | Cat#ABP-BVD-10002 | |||
| RNeasy Plus Mini Kit | Qiagen | Cat#74134 | |||
| CellTrace Violet Proliferation Kit | Invitrogen | Cat#C34557 | |||
| Bio-Safe Coomassie G-250 Stain, 5L | Biorad | Cat#1610787 | |||
| UltraComp eBeads | eBiosciences | Cat#01–2222-42 | |||
| 16% Paraformaldehyde aqueous solution | Electron Microscopy Sciences | Cat#15710 | |||
| EndoH | Produced in house | N/A | |||
| Kifunensine | Toronto Research Chemicals | Cat#K450000 | |||
| BirA | Produced in house (Fairhead and Howarth, 2015) | N/A | |||
| Streptavidin | Sigma | Cat#189730 | |||
| Alexa Flour 647 C2 Maleimide | Invitrogen | Cat#A20347 | |||
| Human MSA-IL-2 | Produced in house (Glassman et al., 2021) | Produced in house | |||
| Human IFNω | Produced in house (Thomas et al., 2011) | Produced in house | |||
| Human IL-15 | R&D systems | Cat#247-ILB-025 | |||
| Human IL-7 | R&D systems | Cat#207-IL-010 | |||
| PE-AnnexinV | Biolegend | Cat#640947 | |||
| Ionomycin | Sigma | Cat#I9657–1MG | |||
| Phorbol 12-myristate 13-acetate (PMA) | Sigma | Cat#P8139–5MG | |||
| Cellfectin II | Gibco | Cat#10362100 | |||
| Carboxypeptidase A | Sigma | Cat#C9268 | |||
| Carboxypeptidase B | Sigma | Cat#217356 | |||
| Expi293 Expression Medium | Gibco | Cat#A1435101 | |||
| Sf-900 III Media | Invitrogen | Cat#12658019 | |||
| ESF 921 Insect Cell Culture Medium | Expression Systems | Cat#96–001-01 | |||
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| MY194-R | This paper | N/A | |||
| MY195-F | This paper | N/A | |||
| MY195-R | This paper | N/A | |||
| IL2Rβ-Nb6 | This paper | N/A | |||
| γc-Nb6 | This paper | N/A | |||
| Human IL2Rβ ECD | This paper | N/A | |||
| Human γc ECD | This paper | N/A | |||
| HIS1 | This paper | N/A | |||
| HIS2 | This paper | N/A | |||
| HIS3 | This paper | N/A | |||
| HIS4 | This paper | N/A | |||
| HIS5 | This paper | N/A | |||
| HIS6 | This paper | N/A | |||
| HIS7 | This paper | N/A | |||
| A1 VHH monomer | This paper | N/A | |||
| 10Rβ1–2Rβ6 (8a.a. linker) | This paper | N/A | |||
| 10Rβ3–2Rβ6 (8a.a. linker) | This paper | N/A | |||
| 10Rβ4–2Rβ1 (8a.a. linker) | This paper | N/A | |||
| 2Rβ3–10Rβ1 (8a.a. linker) | This paper | N/A | |||
| 10Rβ1–2Rβ6 (0a.a. linker) | This paper | N/A | |||
| 10Rβ1–2Rβ6 (4a.a. linker) | This paper | N/A | |||
| 10Rβ1–2Rβ6 (12a.a. linker) | This paper | N/A | |||
| 10Rβ1–2Rβ6 (16a.a. linker) | This paper | N/A | |||
| 10Rβ1–2Rβ6-Fc | This paper | N/A | |||
| Critical Commercial Assays | |||||
| Superdex 200 Increase column 10/300 GL | Cytiva | Cat#28990944 | |||
| Superdex 75 Increase column 10/300 GL | Cytiva | Cat#29148721 | |||
| SA sensor chip | Cytiva | Cat#BR-1005–31 | |||
| HBS-P | Cytiva | Cat#BR-1006–71 | |||
| Yeast-Display Nanobody Library (NbLib) | Kerafast | Cat#EF0014-FP | |||
| Deposited Data | |||||
| γc:γc-Nb6 Complex Crystal Structure | RSCB | 7S2R | |||
| RNA-seq dataset | GEO | GSE183436 | |||
| IL2Rβ:β-Nb6 Complex Crystal Structure | RSCB | 7S2S | |||
| Experimental Models: Cell Lines | |||||
| Human: Expi293F | GIBCO | Cat#A14527 | |||
| Insect: Spodoptera frugiperda (Sf9) | ATCC | Cat#CRL-1711 | |||
| Insect: Trichoplusia ni (T. ni) | Expression Systems | Cat#94–002F | |||
| Human: YT-1 (CD25+) | N/A | ||||
| Human: K-562 | ATCC | Cat#CCL-243 | |||
| Human: A-375 | ATCC | Cat#CRL-1619 | |||
| Recombinant DNA | |||||
| DNA | |||||
| pD649 MY140-F | This paper | N/A | |||
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| pD649 MY141-F | This paper | N/A | |||
| pD649 MY141-R | This paper | N/A | |||
| pD649 MY177-F | This paper | N/A | |||
| pD649 MY177-R | This paper | N/A | |||
| pD649 MY171-F | This paper | N/A | |||
| pD649 MY171-R | This paper | N/A | |||
| pD649 MY144-F | This paper | N/A | |||
| pD649 MY144-R | This paper | N/A | |||
| pD649 MY145-F | This paper | N/A | |||
| pD649 MY145-R | This paper | N/A | |||
| pD649 MY178-F | This paper | N/A | |||
| pD649 MY178-R | This paper | N/A | |||
| pD649 MY172-F | This paper | N/A | |||
| pD649 MY172-R | This paper | N/A | |||
| pD649 MY142-F | This paper | N/A | |||
| pD649 MY142-R | This paper | N/A | |||
| pD649 MY143-F | This paper | N/A | |||
| pD649 MY143-R | This paper | N/A | |||
| pD649 MY179-F | This paper | N/A | |||
| pD649 MY179-R | This paper | N/A | |||
| pD649 MY173-F | This paper | N/A | |||
| pD649 MY173-R | This paper | N/A | |||
| pD649 MY188-F | This paper | N/A | |||
| pD649 MY188-R | This paper | N/A | |||
| pD649 MY189-F | This paper | N/A | |||
| pD649 MY189-R | This paper | N/A | |||
| pD649 MY190-F | This paper | N/A | |||
| pD649 MY190-R | This paper | N/A | |||
| pD649 MY191-F | This paper | N/A | |||
| pD649 MY191-R | This paper | N/A | |||
| pD649 MY192-F | This paper | N/A | |||
| pD649 MY192-R | This paper | N/A | |||
| pD649 MY193-F | This paper | N/A | |||
| pD649 MY193-R | This paper | N/A | |||
| pD649 MY194-F | This paper | N/A | |||
| pD649 MY194-R | This paper | N/A | |||
| pD649 MY195-F | This paper | N/A | |||
| pD649 MY195-R | This paper | N/A | |||
| pD649 IL2Rβ-Nb6 | This paper | N/A | |||
| pD649 γc-Nb6 | This paper | N/A | |||
| pD649 | ATUM | Cat#PD649 | |||
| Software and Algorithms | |||||
| FlowJo v10.5 | Tree Star | RRID: SCR_008520 | |||
| GraphPad Prism 9.1.0 | GraphPad Software | RRID: SCR_002798 | |||
| BIAevaluation software | Cytiva | RRID: SCR_015936 | |||
| PHENIX | (Liebschner et al., 2019) | RRID:SCR_014224 | |||
| Coot | (Emsley et al., 2010) | RRID:SCR_014222 | |||
| SBGrid | (Morin et al., 2013) | RRID:SCR_003511 | |||
| DESEQ2 | (Love et al., 2014) | RRID:SCR_015687 | |||
| STAR | https://github.com/alexdobin/STAR/releases | RRID:SCR_004463 | |||
| UCSF ChimeraX | https://www.cgl.ucsf.edu/chimerax/ | RRID:SCR_015872 |