KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Purified anti-mouse CD16/32 antibody (Clone# 93) | BioLegend | Cat#101302; RRID: AB_312801 |
| CD45 Antibody (Clone# 30–F11), Brilliant Violet 650™ | BioLegend | Cat#103151; RRID: AB_2565884 |
| CD45 Antibody (Clone# 30–F11), Brilliant Violet 785™ | BioLegend | Cat#103149; RRID: AB_2564590 |
| Siglec-F Antibody (Clone# E50–2440), Alexa Fluor® 647 | BD Biosciences | Cat#562680; RRID: AB_2687570 |
| IL-5 Antibody (Clone# TRFK5), APC | BD Biosciences | Cat#554396; RRID: AB_398548 |
| IL-4 Monoclonal Antibody (Clone# 11B11), PE | eBioscience | Cat#12–7041–82; RRID: AB_466156 |
| IL-13 Monoclonal Antibody (Clone# eBio13A), PE-Cyanine7 | eBioscience | Cat#25–7133–82; RRID: AB_2573530 |
| IL-13 Monoclonal Antibody (Clone# eBio13A), PerCP-eFluor 710 | eBioscience | Cat#46–7133–82; RRID: AB_11218496 |
| IFN-γ Antibody (Clone# XMG1.2), Brilliant Violet 605™ | BioLegend | Cat#505840; RRID: AB_2734493 |
| CD4 Antibody (Clone# RM4–5), V500 | BD Biosciences | Cat#560782; RRID: AB_1937327 |
| CD4 Antibody (Clone# GK1.5), Brilliant Violet 605™ | eBioscience | Cat#100451; RRID: AB_2564591 |
| CD4 Antibody (Clone# GK1.5), Alexa Fluor® 700 | BioLegend | Cat# 100430; RRID: AB_493699 |
| CD45RB Antibody (Clone# C363.16A), APC | eBioscience | Cat# 17–0455–81; RRID: AB_2565431 |
| CD25 Monoclonal Antibody (PC61.5), PE-Cyanine7 | eBioscience | Cat# 25–0251–82; RRID: AB_469608 |
| CD11b Antibody (Clone# RM4–5), Brilliant Violet 605™ | BioLegend | Cat#101257; RRID: AB_2565431 |
| CD11b Monoclonal Antibody (Clone# M1/70), eFluor™ 450 | eBioscience | Cat#48–0112–82; RRID: AB_1582236 |
| CD103 Antibody (Clone# 2E7), Brilliant Violet 785™ | BioLegend | Cat#121439; RRID: AB_2800588 |
| Purified anti mouse IL-25 antibody (Clone# 35B) | BioLegend | Cat#514401; RRID: AB_2043954 |
| Purified anti mouse TSLP antibody (Clone# 28F12) | BioLegend | Cat#515202; RRID: AB_2561323 |
| CD40 Antibody (Clone# 3/23), PE/Cyanine5 | BioLegend | Cat#124618; RRID: AB_2075922 |
| CD40 Antibody (Clone# 1C10), PE | eBioscience | Cat#12–0401–82; RRID: AB_465649 |
| KLRG1 (MAFA) Antibody (Clone# 2F1/KLRG1), Brilliant Violet 711™ | BioLegend | Cat#138427; RRID: AB_2629721 |
| CD45R/B220 Antibody (Clone# RA3–6B2), PerCP | BioLegend | Cat#103234; RRID: AB_893353 |
| CD45R (B220) Monoclonal Antibody (Clone# RA3–6B2), eFluor™ 450 | eBioscience | Cat#48–0452–82; RRID: AB_1548761 |
| CD49a Monoclonal Antibody (Clone# HMα1), PE | BioLegend | Cat#142604; RRID: AB_10945158 |
| CD49b Monoclonal Antibody (Clone# HMα2), PE/Cyanine7 | BioLegend | Cat#103518; RRID: AB_2566103 |
| NK-1.1 Monoclonal Antibody (Clone# PK136), Brilliant Violet 605 | BioLegend | Cat#108753; RRID: AB_2686977 |
| CD335 (NKp46) Monoclonal Antibody (Clone# 29A1.4), Brilliant Violet 711 | BioLegend | Cat#137621; RRID: AB_2563289 |
| T1/ST2 (IL-33 R) Monoclonal Antibody (Clone# DJ8), PE | MD Bioproducts | Cat#101001PE |
| T1/ST2 (IL-33 R) Monoclonal Antibody (Clone# DJ8), FITC | MD Bioproducts | Cat#101001F |
| CD127 Monoclonal Antibody (Clone# A7R34), FITC | eBioscience | Cat#11–1271–82; RRID: AB_465195 |
| CD127 Monoclonal Antibody (Clone# A7R34), Alexa Fluor 700 | eBioscience | Cat#56–1271–82; RRID: AB_657611 |
| CD127 Monoclonal Antibody (Clone# eBioRDR5), PE-Cyanine7 | eBioscience | Cat#25–1278–42; RRID: AB_1659672 |
| CD11c Monoclonal Antibody (Clone #N418), eFluor 450 | eBioscience | Cat#48–0114–82; RRID: AB_1548654 |
| CD3e Monoclonal Antibody (Clone# 145–2C11), eFluor™ 450 | eBioscience | Cat#48–0031–82; RRID: AB_10735092 |
| TCR gamma/delta Monoclonal Antibody (Clone# eBioGL3), eFluor™ 450 | eBioscience | Cat#48–5711–82; RRID: AB_2574071 |
| CD5 Monoclonal Antibody (Clone 53–7.3), eFluor™ 450 | eBioscience | Cat#48–0051–80; RRID: AB_1603252 |
| Ly-6G/Ly-6C Monoclonal Antibody (Clone# RB6–8C5), eFluor™ 450 | eBioscience | Cat#48–5931–82; RRID: AB_1548788 |
| Ly-6G/Ly-6C Monoclonal Antibody (Clone# RB6–8C5), APC-eFluor 780 | eBioscience | Cat#47–5931–82; RRID: AB_1518804 |
| FcεRIα Monoclonal Antibody (Clone# MAR-1), Pacific Blue | BioLegend | Cat#134314; RRID: AB_10613298 |
| FOXP3 Monoclonal Antibody (Clone# FJK-16s), eFluor™ 450 | eBioscience | Cat#48–5773–82; RRID: AB_1518812 |
| FOXP3 Monoclonal Antibody (Clone# FJK-16s), PE | eBioscience | Cat#12–5773–82; RRID: AB_465936 |
| FOXP3 Monoclonal Antibody (Clone# MF23), R718 | BD Biosciences | Cat#567095 |
| Gata3 Monoclonal Antibody (Clone# TWAJ), Alexa Fluor™ 488 | eBioscience | Cat#53–9966–42; RRID: AB_2574493 |
| Gata3 Monoclonal Antibody (Clone# TWAJ), eFluor™ 660 | eBioscience | Cat#50–9966–42; RRID: AB_10596663 |
| Gata3 Monoclonal Antibody (Clone# TWAJ), PE-eFluor™ 610 | eBioscience | Cat#61–9966–42; RRID: AB_2574686 |
| ROR gamma (t) Monoclonal Antibody (Clone# B2D), PE-eFluor 610 | eBioscience | Cat#61–6981–82; RRID: AB_2574650 |
| ROR gamma (t) Monoclonal Antibody (Clone# Q31–378), BV650 | BD Biosciences | Cat#564722; RRID: AB_2738915 |
| T-bet Monoclonal Antibody (Clone# eBio4B10 (4B10)), PE-Cyanine7 | eBioscience | Cat#25–5825–82; RRID: AB_11042699 |
| Anti-CD3e (Clone 2C11) | Harlan | Lot#A2022931 |
| Anti-CD28 (Clone 37.51) | Harlan | Lot#A2022929 |
| Anti-IFN-γ (Clone XMG1.2) | Harlan | Lot#A2022930 |
| Anti-IL-4 (Clone 11B.11) | Harlan | Lot#A2022930 |
| Anti-CD16/CD32 (Clone 2.4G2) | Harlan | Lot#A2083015 |
| Dynabeads™ Mouse T-Activator CD3/CD28 for T cell Expansion and Activation | ThermoFisher | Cat#11452D |
| Chemicals, peptides, and recombinant proteins | ||
| Peptide: 2W1S (EAWGALANWAVDSA) | GenScript | Cat#SC1208 |
| Tetramer: 2W1S: IAb (I-A(b): EAWGALANWAVDSA-PE) | NIH Tetramer Facility | N/A |
| Albumin from chicken egg white | Sigma-Aldrich | Cat#A5503 |
| Imject™ Alum Adjuvant | ThermoFisher | Cat#77161 |
| Papain, Carica papaya | Sigma-Aldrich | Cat#5125 |
| UltraComp eBeads™ Plus Compensation Beads | eBioscience | Cat#01–3333–42 |
| Recombinant Mouse IL-1 β | R&D Systems | Cat#401-ML |
| Recombinant Mouse IL-2 protein | R&D Systems | Cat#402-ML |
| Recombinant Mouse IL-4 Protein | R&D Systems | Cat#404-ML |
| Recombinant Mouse IL-6 Protein | PeproTech | Cat#216–16 |
| Recombinant Mouse IL-7 Protein | R&D Systems | Cat#407–ML–005 |
| Recombinant Mouse IL-12 Protein | PeproTech | Cat#210–12 |
| Recombinant Mouse IL-13 Protein | R&D Systems | Cat#413–ML–005 |
| Recombinant Mouse IL-25 (IL-17E) | BioLegend | Cat#587302 |
| Recombinant Mouse IL-33 Protein | R&D Systems | Cat#3626–ML |
| Recombinant Mouse TSLP Protein | R&D Systems | Cat#555–TS–010/CF |
| Recombinant Mouse TGF-P1 Protein | R&D Systems | Cat#7666–MB |
| SYBR™ Safe DNA Gel Stain | ThermoFisher | Cat#S33102 |
| Platinum™ Hot Start PCR Master Mix (2X) | ThermoFisher | Cat#13000012 |
| DNA Gel Loading Dye | ThermoFisher | Cat#R0611 |
| 100 bp DNA Ladder | ThermoFisher | Cat#15628019 |
| HBSS, calcium, magnesium | ThermoFisher | Cat#24020117 |
| ACK Lysing Buffer | ThermoFisher | Cat#A1049201 |
| Collagenase, Type IV, powder | ThermoFisher | Cat#17104019 |
| DNase I recombinant grade I, from bovine pancreas, expressed in Pichia pastoris | Roche | Cat#4536282001 |
| PBS, pH 7.4 | ThermoFisher | Cat#10010031 |
| Albumin, Bovine Fraction V | MP | Cat#160069 |
| RPMI 1640 Medium | ThermoFisher | Cat#21870–076 |
| Fetal Bovine Serum | R&D Systems | Cat#S12450 |
| Penicillin-Streptomycin | ThermoFisher | Cat#15140122 |
| L-Glutamine (200 mM) | ThermoFisher | Cat#25030081 |
| MEM Non-Essential Amino Acids Solution (100X) | ThermoFisher | Cat#11140050 |
| Sodium Pyruvate | ThermoFisher | Cat#11360 |
| 2-Mercaptoethanol | ThermoFisher | Cat#21985023 |
| E-Gel™ SizeSelect™ II Agarose Gels, 2% | ThermoFisher | Cat#G661012 |
| Nystatin | Sigma-Aldrich | Cat#N6261–500KU |
| Activated charcoal | Sigma-Aldrich | Cat#C2764–500G |
| Phorbol 12-myristate 13-acetate | Sigma-Aldrich | Cat#P8139–5MG |
| Ionomycin | Sigma-Aldrich | Cat#407952–5MG |
| Brefeldin A Solution | BioLegend | Cat#420601 |
| Qiazol lysis reagent | QIAGEN | Cat#79306 |
| DMSO | Sigma-Aldrich | Cat#D2650 |
| 1000 ML PREFILLED FORMALIN CONTAINER | Azer Scientific | Cat#PFNBF-1000 |
| eBioscience™ Foxp3/Transcription Factor Staining Buffer Set | eBioscience | Cat#00–5523–00 |
| Fixable Viability Dye eFluor™ 780 | eBioscience | Cat#65–0865–14 |
| Critical commercial assays | ||
| Mouse IL-17E Duoset ELISA | R&D Systems | Cat#DY1399 |
| Mouse IL-33 DuoSet ELISA | R&D Systems | Cat#DY3626–05 |
| Mouse TSLP Quantikine ELISA kit | R&D Systems | Cat#MTLP00 |
| Qubit™ RNA High Sensitivity (HS), Assay Kit | ThermoFisher | Cat#Q10210 |
| Qubit™ dsDNA Broad Rang (BR) Assay Kit | ThermoFisher | Cat#Q32850 |
| RNeasy Micro Kit (50) | QIAGEN | Cat#74004 |
| MinElute Gel Extraction Kit (50) | QIAGEN | Cat#28604 |
| MinElute Reaction Cleanup Kit | QIAGEN | Cat#28206 |
| RNase-Free DNase Set | QIAGEN | Cat#79256 |
| Ovation® RNA-Seq System V2 | TECAN | Cat#7102–32 |
| KAPA LTP Library Preparation Kit | Roche | Cat#KK8232 |
| Naive CD4 T cell isolation Kit mouse | Miltenyi Biotec | Cat#130–104–453 |
| EasySep™ Mouse ILC2 Enrichment Kit | STEMCELL | Cat#19842 |
| Deposited data | ||
| RNA-Seq Raw and analyzed data | This paper | GEO: GSE166779 |
| Experimental models: Organisms/strains | ||
| Mouse: KIrg1Cre; strain background: C57BL/6 | Dr. Richard Flavell of Yale University | Herndler-Brandstetter et al.27 |
| Mouse: Gata3fl/fl; strain background: C57BL/6 | NIAID-Taconic repository | Zhu et al.14 (Line 355) |
| Mouse: hCD2Cre; strain background: C57BL/6 | Dr. Paul Love of NICHD, NIH | Vacchio et al.51 (also available as JAX line: 027406) |
| Mouse: Crlf2−/−;strain background: C57BL/6 | Dr. Warren J. Leonard of NHLBI, NIH | Al-Shami et al.36 |
| Mouse: IL-33 KO (IL-33 trap); strain background: C57BL/6 | Late Dr. William E. Paul of NIAID, NIH | Pichery et al.35 |
| Mouse: CIKS KO; strain background: C57BL/6 | NIAID-Taconic repository | Line: 290 |
| Mouse: Rag1−/−; B6.SJL-[KO]RAG1 | NIAID-Taconic repository | Line: 165 |
| Mouse: Rag2−/−II2rg−/−; C57BL/10SgSnAi-[KO]RAG2-[KO]gamma c | NIAID-Taconic repository | Line: 111 |
| Mouse: OT-II; B6.SJL-Ptprca Rag1tm1 Tg(TcraTcrb)425Cbn | NIAID-Taconic repository | Line: 361 |
| Mouse: C57BL/6 (B6NTac) | Taconic | C57BL/6NTac |
| Mouse: II4−/−; strain background: C57BL/6 | NIAID-Taconic repository | Line: 46 |
| Mouse: II4/II13−/−; strain background: C57BL/6 | NIAID-Taconic repository | Line: 242 |
| Oligonucleotides | ||
| please see Table S1 for Oligo sequences | IDT | N/A |
| Software and algorithms | ||
| FlowJo V10 | FlowJo LLC | https://www.flowjo.com |
| Partek Flow | Partek | https://www.partek.com |
| GraphPad Prism | GraphPad | https://www.graphpad.com |