Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (M. musculus) | Rag-deficient B6.129S7-Rag < tm1mom > JAusb (RAG1) |
Australian BioResources | Used for in vivo experiments | |
| Strain, strain background (M. musculus) | B6.SJL-PtprcaPepcb/BoyJAusb (PTPRCA) |
Australian BioResources | Used for in vivo experiments | |
| Strain, strain background (M. musculus) | B6.129p2-ccr5 tm1Kuz/J (CCR5-/-) |
Australian BioResources | 7–12 week mice used to isolate CCR5KO CTLs | |
| Strain, strain background (M. musculus) | B6-Tg-OTI (OT1) |
Australian BioResources | 7–12 week mice used to isolate OT1 CTLs | |
| Strain, strain background (M. musculus) | B6-Tg(CAG-Lifeact/EGFP)-OTI (OT1GFP) |
Galeano Niño et al., 2020 | 7–12 week mice used to isolate LifeactGFP OT1 CTLs | |
| Strain, strain background (M. musculus) | B6.gBT-I/uGFP (gBT1-GFP) |
Gift from W. R. Heath | 7–12 week mice used to isolate GFP gBT1 CTLs | |
| Strain, strain background (M. musculus) | B6.gBT-I/dsRED (gBT1-dsRED) |
Gift from W. R. Heath | 7–12 week mice used to isolate dsRED gBT1 CTLs | |
| Cell line (M. musculus) | EL-4 lymphoma | ATCC | Cat. #: TIB-39 | |
| Cell line (M. musculus) | NIH/3T3 fibroblast | ATCC | Cat. #: CRL-1658 | |
| Cell line (Homo-sapiens) | WK1 glioblastoma cells | Day et al., 2013 | Cultured as described in Stringer et al., 2019 | |
| Cell line (Homo-sapiens) | GP2-293 embryonic kidney cells | Clonetech | Cat. #: 631458 | |
| Cell line (M. musculus) | mCCL3-mScarletI secreting EL4 cells | This paper | EL4 cells transduced with CCL3-mScarletI construct. Sequence available in Materials and methods. | |
| Cell line (M. musculus) | mCCL4-miRFP670 secreting EL4 cells | This paper | EL4 cells transduced with CCL4-miRFP670 construct. Sequence available in Materials and methods. | |
| Cell line (M. musculus) | ‘CCL3/4-secreting EL4’ i.e. mCCL3-mScarletI and mCCL4-miRFP670 secreting EL4 cells | This paper | EL4 cells transduced with CCL3-mScarletI and CCL4-miRFP670 construct. Sequences available in Materials and methods. | |
| Cell line (M. musculus) | EL4.HSV | This paper | EL4 cells transduced with bicistronic SSIEFARL epitope and mTagBFP2. Sequence available in Materials and methods. | |
| Cell line (M. musculus) | EL4.OVA | This paper | EL4 cells transduced with bicistronic SIINFEKL epitope and mTagBFP2. Sequence available in Materials and methods. | |
| Cell line (Homo-sapiens) | CAR T cells | This paper | Primary human T cells transduced with EphA2-specific CAR construct. Produced by Kramer lab at Children's Cancer Research Unit, The Children's Hospital at Westmead. Information on CAR constructs available in Materials and methods. | |
| Transfected construct (M. musculus) | MSCV-based retrovial expression vector (LENC) |
Fellmann et al., 2013
Gift from J. Zuber |
http://doi.org/10.1016/j.celrep.2013.11.020
|
Retrovirus construct to express cDNA |
| Transfected construct (M. musculus) | pMD2.G | Gift from Didier Trono | Addgene plasmid # 12259; RRID:Addgene_12259 | Construct for VSVG pseudotyping retrovirus |
| Transfected construct (M. musculus) | mCCL3-mScarletI gBlocks | Integrated DNA Technologies | gBlocks cloned into LENC | |
| Transfected construct (M. musculus) | mCCL4-miRFP670 gBlocks | Integrated DNA Technologies | gBlocks cloned into LENC | |
| Transfected construct (M. musculus) | SSIEFARL-mTagBFP2 gBlocks | Integrated DNA Technologies | gBlocks cloned into LENC | |
| Transfected construct (M. musculus) | SIINFEKL-mTagBFP2 gBlocks | Integrated DNA Technologies | gBlocks cloned into LENC | |
| Transfected construct (Homo- sapiens) | EphA2-specific CAR construct | Chow et al., 2013 and Yi et al., 2018 | Construct for lentiviral transduction of human T cells | |
| Biological sample (Homo-sapiens) | Peripheral blood mononuclear cells (PBMCs) | Sydney Children’s Hospitals Network | Used to isolate human T cells | |
| Antibody | Anti-XCL1 (polyclonal goat IgG) | R & D Systems | Cat. #: AF486 RRID:AB_2216915 |
(5 µg/ml) |
| Antibody | Anti-CCL1 (monoclonal rat IgG2a) | R & D Systems | Cat. #: MAB845 RRID:AB_2070618 |
(5 µg/ml) |
| Antibody | Anti-CXCR2 (monoclonal rat IgG2a) | R & D Systems | Cat. #: MAB2164-100 RRID:AB_358062 |
(10 µg/ml) |
| Antibody | Anti-CCL3 (polyclonal goat IgG) | R & D Systems | Cat. #: AF-450-NA RRID:AB_354492 |
(5 µg/ml) |
| Antibody | Anti-CCL4 (monoclonal rat IgG2a) | R & D Systems | Cat. #: MAB451-100 RRID:AB_2259676 |
(5 µg/ml) |
| Antibody | Anti-CXCR4 (monoclonal rat IgG2b) | R & D Systems | Cat. #: MAB21651-100 RRID:AB_2801441 |
(10 µg/ml) |
| Antibody | Anti-CCL5 (polyclonal goat IgG) | R & D Systems | Cat. #: AF478 RRID:AB_10080077 |
(1 µg/ml) |
| Antibody | Anti-CCL9/10 (monoclonal rat IgG1) | R & D Systems | Cat. #: MAB463 RRID:AB_2259783 |
(5 µg/ml) |
| Antibody | Anti-CXCL10 (monoclonal rat IgG2a) | R & D Systems | Cat. #: MAB466 RRID:AB_2292486 |
(1 µg/ml) |
| Antibody | Anti-CXC12 (monoclonal mouse IgG1) | R & D Systems | Cat. #: MAB310 RRID:AB_2276927 |
(1 µg/ml) |
| Antibody | Anti-CD3ε (monoclonal Armenian Hamster) | BioLegend | Cat. #: 100302 RRID:AB_312667 |
(1 µg/ml) |
| Antibody | Anti-CD28 (monoclonal Syrian Hamster) | BioLegend | Cat. #: 102101 RRID:AB_312866 |
(1 µg/ml) |
| Antibody | Rat IgG1, κ Isotype control (Rat monoclonal) | BioLegend | Cat. #: 400402 RRID:AB_326508 |
(1 µg/ml) |
| Antibody | Mouse IgG1, κ Isotype control (Mouse monoclonal) | BioLegend | Cat. #: 401401 RRID:AB_2801452 |
(1 µg/ml) |
| Antibody | APC anti-CD44 (monoclonal rat IgG2b) | BD Biosciences | Cat. #: 553991 RRID:AB_10050405 |
(1 µg/ml) |
| Antibody | Pacific Blue anti-CD8a (monoclonal rat IgG2a) | BD Biosciences | Cat. #: 558106 RRID:AB_397029 |
(1 µg/ml) |
| Antibody | Biotin anti-CD3ε (monoclonal Armenian Hamster) | Thermo Fisher Scientific | Cat. #: 13-0031-82 RRID:AB_466319 |
(1 µg/ml) |
| Antibody | Biotin anti-CD28 (monoclonal Syrian Hamster) | Thermo Fisher Scientific | Cat. #: 13-0281-82 RRID:AB_466411 |
(1 µg/ml) |
| Antibody | Biotin anti-CD3 (monoclonal mouse IgG2a) | Thermo Fisher Scientific | Cat. #: 13-0037-82 RRID:AB_1234955 |
(1 µg/ml) |
| Antibody | Biotin anti-CD28 (monoclonal mouse IgG1) | Thermo Fisher Scientific | Cat. #: 13-0289-82 RRID:AB_466415 |
(1 µg/ml) |
| Antibody | Biotin rat IgG2a κ Isotype Control (Rat polyclonal) | eBioscience | Cat. #: 13-4321-82 RRID:AB_470084 |
(1 µg/ml) |
| Antibody | Biotin IgG Isotype Control (Armenian hamster) | eBioscience | Cat. #: 14-4888-81 RRID:AB_470128 |
(1 µg/ml) |
| Antibody | Brilliant Violet 711 anti-CD11b (monoclonal rat IgG2b) | BioLegend | Cat. #: 101241 RRID:AB_11218791 |
(2 µg/ml) |
| Antibody | Alexa Fluor 488 anti-NK1.1 (monoclonal mouse IgG2a) | BioLegend | Cat. #: 108717 RRID:AB_493184 |
(2 µg/ml) |
| Antibody | FITC anti-Ly-6C (monoclonal rat IgG2c) | BioLegend | Cat. #: 128005 RRID:AB_1186134 |
(2 µg/ml) |
| Antibody | Brilliant Violet 510 anti-CD90.2 (monoclonal rat IgG2b) | BioLegend | Cat. #: 105335 RRID:AB_2566587 |
(2 µg/ml) |
| Antibody | Brilliant Violet 421 anti-CD64 (monoclonal mouse IgG1) | BioLegend | Cat. #: 139309 RRID:AB_AB_2562694 |
(2 µg/ml) |
| Antibody | APC/Fire 750 anti-CD45 (monoclonal rat IgG2b) | BioLegend | Cat. #: 103153 RRID:AB_2572115 |
(2 µg/ml) |
| Antibody | APC/Fire 750 anti-CD45.1 (monoclonal mouse IgG2a) | BioLegend | Cat. #: 110751 RRID:AB_2629805 |
(2 µg/ml) |
| Antibody | FITC anti-CD11c (monoclonal Armenian Hamster) | BioLegend | Cat. #: 117305 RRID:AB_313774 |
(2 µg/ml) |
| Antibody | Pacific blue anti-MHC class II (monoclonal rat IgG2b) | BioLegend | Cat. #: 107619 RRID:AB_493528 |
(2 µg/ml) |
| Antibody | APC anti-CD8a (monoclonal rat IgG2a) | BioLegend | Cat. #: 100711 RRID:AB_312750 |
(2 µg/ml) |
| Antibody | Biotin anti-CCL3 (polyclonal Rabbit sera) | PeproTech | Cat. #: 500-P121BT | (1 µg/ml) |
| Peptide, recombinant protein | OVA257-264 peptide, SIINFEKL | AusPep | Cat. #: 2711 | (1, 0.2, 0.1, 0.02, 0.001 µg/ml) |
| Peptide, recombinant protein | gB498-505 peptide, SSIEFARL | Auspep | Cat. #: gb498-505 | (1 µg/ml) |
| Peptide, recombinant protein | Recombinant mouse IL-2 | BioLegend | Cat. #: 575408 | (100 ng/ml) |
| Peptide, recombinant protein | Recombinant human IL-7 | Miltenyi Biotech Australia | Cat. #: 130-095-362 | (10 ng/ml) |
| Peptide, recombinant protein | Recombinant human IL-15 | Miltenyi Biotech Australia | Cat. #: 130-095-764 | (5 ng/ml) |
| Peptide, recombinant protein | Corning Collagen I Rat Tail Natural | In Vitro Technologies | Cat. #: FAL354236 | (~1.5 mg/ml) |
| Peptide, recombinant protein | Monobiotinylated H-2Kb/SIINFEKL | Tetramer Synthesis Service | John Curtin School of Medical Research | (1 µg/ml) |
| Peptide, recombinant protein | Recombinant human (cross reactive with murine) CCL1 | Peprotech | Cat. #: 300–37 | (1, 10 and 100 ng/ml) |
| Peptide, recombinant protein | Recombinant Mouse XCL1 | R & D systems | Cat. #: 486-LT-025 | (1, 10 and 100 ng/ml) |
| Peptide, recombinant protein | Recombinant Murine CCL3 | Peprotech | Cat. #: 250–09 | (1, 10 and 100 ng/ml) |
| Peptide, recombinant protein | Recombinant Murine CCL4 | Peprotech | Cat. #: 250–32 | (1, 10 and 100 ng/ml) |
| Peptide, recombinant protein | Recombinant Murine CCL5 | Peprotech | Cat. #: 250–07 | (1, 10 and 100 ng/ml) |
| Peptide, recombinant protein | Recombinant murine CCL9/10 | Peprotech | Cat. #: 250–12 | (1, 10 and 100 ng/ml) |
| Peptide, recombinant protein | Recombinant murine CXCL10 | Peprotech | Cat. #: 250–16 | (1, 10 and 100 ng/ml) |
| Peptide, recombinant protein | Recombinant Human CXCL12 | Peprotech | Cat. #: 300-28A | (1, 10 and 100 ng/ml) |
| Peptide, recombinant protein | Lysyl Endopeptidase, Mass Spectrometry Grade (Lys-C) | FUJIFILM Wako Pure Chemical Corp | Cat. #: 121–05063 | (1 µg) |
| Sequence-based reagent | TruSeq Stranded mRNA | Illumina | Cat. #: 20020594 | |
| Chemical compound, drug | CellTracker Orange 5-(and-6)-(((4-chloromethyl) benzoyl) amino) tetramethylrhodamine (CMTMR) |
Invitrogen | Cat. #: C2927 | (5 µM) |
| Chemical compound, drug | CellTracker Deep Red | Invitrogen | Cat. #: C34565 | (1 µM) |
| Chemical compound, drug | CellTracker Green 5-chloromethylfluorescein diacetate (CMFDA) | Invitrogen | Cat. #: C7025 | (5 µM) |
| Chemical compound, drug | Ketamine | Provet Pty Ltd | Cat. #: KETAM I | (100 mg/kg) |
| Chemical compound, drug | Xylazine | Provet Pty Ltd | Cat. #: XYLA Z2 | (15 mg/kg) |
| Chemical compound, drug | Pertussis toxin from Bordetella pertussis (PTX) | Sigma Aldrich | Cat. #: P7208 | (100 ng/ml) |
| Chemical compound, drug | Inactive mutated version of Pertussis toxin from Bordetella pertussis (m-PTX) | Sigma Aldrich | Cat. #: PT-16.0003 | (100 ng/ml) |
| Chemical compound, drug | Maraviroc (CCR5 antagonist) | Sigma Aldrich | Cat. #: PZ0002-5mg | (10 µg/ml) |
| Chemical compound, drug | Cenicriviroc (CCR2 + CCR5 dual inhibitor) | AdooQ Bioscience | Cat. #: A13632-10 (000–22038) | (1 µM) |
| Chemical compound, drug | CCR2 antagonist | Santa Cruz Biotechnology Inc | Cat. #: sc-202525 | (1 µM) |
| Chemical compound, drug | UCB35625 (CCR1 + CCR3 dual inhibitor) | R & D Systems | Cat. #: 2757/1 | (50 µM) |
| Chemical compound, drug | Iodoacetamide (IAM) | Sigma Aldrich | Cat. #: I6125-5G | (1 M) |
| Chemical compound, drug | Dithiothreitol (DTT) | Sigma Aldrich | Cat. #: 43816–10 ML | (50 mM) |
| Commercial assay or kit | EasySep Direct Human T Cell Isolation Kit | Stem Cell Technologies | Cat. #: 19661 | |
| Commercial assay or kit | T Cell TransAct, human | Miltenyi Biotech Australia | Cat. #: 130-111-160 | |
| Commercial assay or kit | EasySep Human CD19 Positive Selection Kit II | Stem Cell Technologies | Cat. #: 17854 | |
| Commercial assay or kit | RNeasy Mini Kit | Qiagen | Cat. #: 74104 | |
| Commercial assay or kit | High-Capacity cDNA Reverse Transcription Kit | ThermoFisher Scientific | Cat. #: 4368814 | |
| Commercial assay or kit | Sandwich ELISA kits (CCL1) | OriGene Technologies | Cat. #: EA100390 | |
| Commercial assay or kit | Sandwich ELISA kits (CCL9) | OriGene Technologies | Cat. #: EA100725 | |
| Commercial assay or kit | Cytometric Bead Array: Multi-Analyte Flow Assay Kit (CCL3, CCL4, CCL5, CXCL10, TNF-α and IFN-γ) | LEGENDplex | Custom mouse panel | |
| Commercial assay or kit | BD Cytometry Beads array Mouse MIP-1b Flex Set (Bead C9) | BD Biosciences | Cat. #: 558343 | |
| Commercial assay or kit | BD Cytometry Beads array Mouse MIP-1a Flex Set (Bead C7) | BD Biosciences | Cat. #: 558449 | |
| Commercial assay or kit | CellTrace CFSE Proliferation Kit | ThermoFisher Scientific |
Cat. #: C34554 | |
| Commercial assay or kit | CellTrace Violet Proliferation Kit | ThermoFisher Scientific |
Cat. #: C34571 | |
| Commercial assay or kit | Cytofix/Cytoperm Plus Kit (with BD GolgiStop) | BD Bioscience | Cat. #: 554715 | |
| Software, algorithm | FlowJo software version 10.2 | Tree Star Inc | flowjo.com | |
| Software, algorithm | Prism GraphPad Software version 8 | GraphPad | Imaris.oxinst.com | |
| Software, algorithm | Leica Application Suite (LAS) X software version 3.3.0.16794 | Leica Microsystems | Leica-microsystems.com | |
| Software, algorithm | FLUOstaromega software version 5.10 R2 | BMG Labtech GmbH | Bmglabtech.com | |
| Software, algorithm | CFX Manager Software | Bio-Rad | Bio-rad.com | |
| Software, algorithm | BioStation IM software | Nikon | nikon.com | |
| Software, algorithm | ImageJ/FIJI | US National Institutes of Health | Imagej.net | |
| Software, algorithm | Matlab | Mathworks | Mathworks.com | |
| Software, algorithm | R | R Core Team | r-project.org | |
| Software, algorithm | Plot2 for Mac 2.6.1 | Mike Wesemann | apps.micw.org | |
| Software, algorithm | motilisim | Read et al., 2016 | https://github.com/marknormanread/TcellSwarming | |
| Software, algorithm | FastQC (version 0.11.5) | Babraham Institute | bioinformatics.babraham.ac.uk | |
| Software, algorithm | Trimmomatic (version 0.36) | Bolger, A. M., Lohse, M., and Usadel, B. | usadellab.org | |
| Software, algorithm | FeatureCounts (version 1.5.1) | Bioconductor | bioconductor.org | |
| Software, algorithm | STRING DB | ELIXIR | string-db.org | |
| Software, algorithm | MaxQuant (version 1.5.8.3) | Max Plank Insitute of Biochemistry | maxquant.org | |
| Software, algorithm | LFQ Analyst | Monash University | bioinformatics.erc.monash.edu/apps/LFQ-Analyst | |
| Software, algorithm | Limma | Bioconductor | bioconductor.org | |
| Software, algorithm | Motility_analysis package | Python Software Foundation | https://github.com/marknormanread/TcellSwarming | |
| Other | Streptavidin-coated polystyrene particles | Spherotech Inc | Cat. #: SVP-60–5 | 6.0–8.06 µm diameter |
| Other | SPHERO AccuCount blank particles | Spherotech Inc | Cat. #: QACBP-70–10 | |
| Other | Sera-Mag Speed beads | ThermoFisher Scientific | Cat. #: 65152105050250 | |
| Other | Sera-Mag Speed beads | ThermoFisher Scientific | Cat. #: 45152105050250 | |
| Other | Trypsin-gold | Promega | Cat. #: V5280 | |
| Other | Dimethyl sulfoxide (DMSO) | Sigma-Aldrich | Cat. #: D2650-100ml | |
| Other | Collagenase IV | Sigma-Aldrich | Cat. #: C5138-1G | |
| Other | 4’, 6-Diamidino-2-phenylindole (DAPI) | ThermoFisher Scientific | Cat. #: D3571 | (0.5 µg/ml) |
| Other | 10 kDA Dextran labelled with AlexaFluor647 (Dextran-AF647) | ThermoFisher Scientific | Cat. #: D22914 | (10 µg/ml) |
| Other | Fmoc-Asp-Wand resin | GL Biochem | Cat. #: 40501 | |
| Other | 1-hydroxybenzotriazole (HOBt) | GL Biochem | Cat. #: 2592-95-2 | |
| Other | 2-(1H-Benzotriazole-1-yl)−1,1,3,3-Tetramethyluronium hexafluorophosphate (HBTU) | GL Biochem | Cat. #: 94790-37-1 |