KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| eFluor 450 anti-mouse CD4 | ThermoFisher Scientific | Catalog #: 48-0042-82; Clone:RM4-5; RRID:AB_1272194 |
| Brilliant Violet 605 anti-mouse PD1 | Biolegend | Catalog #: 135220; Clone: 29F.1A12; RRID:AB_2562616 |
| Brilliant Violet 650 anti-mouse CD62L | Biolegend | Catalog #: 104453; Clone: MEL-14; RRID:AB_2800559 |
| Brilliant Violet 785 anti-mouse CD45 | Biolegend | Catalog #: 103149; Clone: 30-F11; RRID:AB_2564590 |
| FITC anti-mouse TIM-3 | ThermoFisher Scientific | Catalog #: 11-5870-82; Clone: RMT3-23; RRID:AB_2688129 |
| PerCP-eFluor 710 anti-mouse CD8 alpha | ThermoFisher Scientific | Catalog #: 46-0081-82; Clone: 53-6.7; RRID:AB_1834433 |
| PE anti-mouse LAG-3 | Biolegend | Catalog #: 125208; Clone: C9B7W; RRID:AB_2133343 |
| PE/Dazzle 594 anti-mouse CD44 | Biolegend | Catalog #: 103056; Clone: IM7; RRID:AB_2564044 |
| PE-Cy7 anti-mouse Ki67 | ThermoFisher Scientific | Catalog #: 25-5698-82; Clone: SolA15; RRID:AB_11220070 |
| Alexa Fluor 647 anti-mouse CD3 | Biolegend | Catalog #: 100209; Clone: 17A2; RRID:AB_389323 |
| APC-Cy7 anti-mouse CD19 | Biolegend | Catalog #: 115530; Clone: 6D5; RRID:AB_830707 |
| eFlour 450 anti-mouse IgM | ThermoFisher Scientific | Catalog #: 48-5890-82; Clone: eB121-15F9; RRID:AB_10671539 |
| PE/Dazzle 594 anti-mouse IgD | Biolegend | Catalog #: 405742; Clone: 11-26c.2a; RRID:AB_2571985 |
| PerCP-Cy5.5 anti-mouse CD11c | Biolegend | Catalog #: 117328; Clone: N418; RRID:AB_2129641 |
| PE anti-mouse I-A/I-E (MHC II) | Biolegend | Catalog #: 107608; Clone: M5/114.15.2; RRID:AB_313323 |
| Pacific Blue anti-mouse ICAM-1 (CD54) | Biolegend | Catalog #: 116116; Clone: YN1/1.7.4; RRID:AB_2121771 |
| Brilliant Violet 605 anti-mouse VCAM-1 (CD106) | BD Bioscience | Catalog #: 745193; Clone: 429; RRID:AB_2742789 |
| Alexa Fluor 488 polyclonal Collagen Type I | SoutherBiotech | Catalog #: 1310-30; Clone: Poly; RRID:AB_2794728 |
| PerCP-eFlour 710 anti-mouse CD31 | ThermoFisher Scientific | Catalog #: 46-0311-82; Clone: 390; RRID:AB_1834429 |
| Unconjugated anti-mouse alpha smooth muscle actin | ThermoFisher Scientific | Catalog #: 14-9760-82; Clone: 1A4; RRID:AB_2572996 |
| PE anti-mouse IgG2a | Biolegend | Catalog #: 407108; Clone: RMG2a-62; RRID:AB_10549456 |
| PE/Dazzle 594 anti-mouse CD140 alpha | Biolegend | Catalog #: 135922; Clone: APA5; RRID:AB_2814035 |
| PE-Cy7 anti-mouse podoplanin (gp38) | Biolegend | Catalog #: 127412; Clone: 8.1.1; RRID:AB_10613648 |
| eFlour 660 anti-mouse Ki67 | ThermoFisher Scientific | Catalog #: 50-5698-82; Clone: SolA15; RRID:AB_2574235 |
| Unconjugated anti-mouse FAP | Millipore Sigma | Catalog #: MABC1145; Clone: 73.3: RRID:AB_2892562 |
| APC-Cy7 anti-mouse IgG1 | Biolegend | Catalog #: 406620; Clone: RMG1-1; RRID:AB_2571947 |
| PE/Dazzle 594 anti-mouse PD-L1 | Biolegend | Catalog #: 124324; Clone: 10F.9G2; RRID:AB_2565639 |
| APC anti-mouse CD120 alpha (TNF R Type I/p55) | Biolegend | Catalog #: 113006; Clone: 55R-286; RRID:AB_2208779 |
| Brilliant Violet 421 anti-mouse CD120 beta (TNF R Type II/p75) | BD Bioscience | Catalog #: 564088; Clone: TR75-89; RRID:AB_2738585 |
| PE/Dazzle 594 anti-mouse CD90.2 (Thy-1.2) | Biolegend | Catalog #: 140329; Clone: 53-2.1; RRID:AB_2650965 |
| APC anti-mouse lymphotoxin beta receptor (LTβR) | Biolegend | Catalog #: 134408; Clone: 5G11; RRID:AB_2728151 |
| DyLight 550 Anti-Mouse/Human CD45R (B220) | Leinco Technologies (Discontinued) | Catalog #: C1944; Clone: RA3-6B2; RRID:AB_2829026 |
| Unconjugated anti-mouse Podoplanin (gp38) | Biolegend | Catalog #: 127402; Clone: 8.1.1; RRID:AB_1089187 |
| Biotin anti-mouse/human PNAd | Biolegend | Catalog #: 120804; Clone: MECA-79; RRID:AB_493557 |
| Alexa Fluor 488 anti-mouse CD45 | Biolegend | Catalog #: 103122; Clone: 30-F11; RRID:AB_493531 |
| Biotin anti-mouse ICAM-1 (CD54) | Biolegend | Catalog #: 116103; Clone: YN1/1.7.4; RRID:AB_313694 |
| Alexa Fluor 647 anti-mouse VCAM-1 (CD106) | Biolegend | Catalog #: 105712; Clone: 429; RRID:AB_493429 |
| Alexa Fluor 488 AffiniPure Goat Anti-Syrian Hamster IgG (H+L) | Jackson Labs | Catalog #: 107-545-142; Clone: Poly; RRID:AB_2337478 |
| Rhodamine (TRITC) AffiniPure Goat Anti-Syrian Hamster IgG (H+L) | Jackson Labs | Catalog #: 107-025-142; Clone: Poly; RRID:AB_2337452 |
| Rhodamine (TRITC) AffiniPure Donkey Anti-Mouse IgG (H+L) | Jackson Labs | Catalog #: 715-025-150; Clone: Poly; RRID:AB_2340766 |
| Alexa Fluor 350 Streptavidin | ThermoFisher Scientific | Catalog #: S11249; Clone: N/A |
| InVivoPlus anti-mouse PD-L1 (B7-H1) | BioXcell | Catalog #: BP0101; Clone: 10F.9G2; RRID:AB_10949073 |
| InVivoPlus anti-mouse PD-1 (CD279) | BioXcell | Catalog #: BP0273; Clone: 29F.1A12; RRID:AB_2687796 |
| InVivoPlus anti-mouse CTLA-4 (CD152) | BioXcell | Catalog #: BP0032; Clone: UC10-4F10-11; RRID:AB_1107598 |
| Biotin anti-mouse Podoplanin | Biolegend | Catalog #: 127404; Clone: 8.1.1; RRID:AB_1134222 |
| Biotin anti-mouse CD31 | Biolegend | Catalog #: 102404; Clone: 390; RRID:AB_312899 |
| CD45 MicroBeads, mouse | Miltenyi Biotec | Catalog #: 130-097-153 |
| Anti-Biotin MicroBeads | Miltenyi Biotec | Catalog #: 130-097-046 |
| InVivomAb anti-mouse CD16/CD32 | BioXcell | Catalog #: BE0307; Clone: 2.4G2; RRID:AB_2736987 |
| Unconjugated anti-mouse/human Lymphotoxin alpha (LTA) | Abcam | Catalog #: ab100844; Clone: AT15A3; RRID:AB_10672573 |
| Unconjugated anti-human VCAM-1 | Nordic Biosite | Catalog #: LS-C391116-20; Clone: 1.4C3; RRID:AB_2892563 |
| Unconjugated anti-human CXCL13 | Novus Biological | Catalog #: NBP2-16041; Clone: Poly; RRID:AB_2892564 |
| Unconjugated anti-human/Rat/Mouse APRIL | Abcam | Catalog #: ab200836; Clone: EPR14588; RRID:AB_2892565 |
| Unconjugated anti-human gp36 | Abcam | Catalog #: ab236529; Clone: EPR22182; RRID:AB_2889051 |
| Unconjugated anti-human CD20 | Agilent | Catalog #: M075501-2; Clone: L26; RRID:AB_2892566 |
| Unconjugated anti-mouse CD4 | Abcam | Catalog #: ab183685; Clone: EPR19514; RRID:AB_2686917 |
| Unconjugated anti-mouse CD8 | Abcam | Catalog #: ab237723; Clone: CAL38; RRID:AB_2864723 |
| Unconjugated anti-mouse CD19 | Cell Signaling Technology | Catalog #: 90176S; Clone: D4V4B; RRID:AB_2800152 |
| Unconjugated anti-human/rat/mouse CD34 | Abcam | Catalog #: ab81289; Clone: EP373Y; RRID:AB_1640331 |
| Unconjugated anti-mouse CD11c | Abcam | Catalog #: ab219799; Clone: EPR21826; RRID:AB_2864725 |
| Biological samples | ||
| Melanoma tissue samples from patients | This paper | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Recombinant LTβR-Ig fusion protein | Wu etal. (1999) J. Exp. Med. 190, 629 | N/A |
| DNase I | Sigma | Catalog #: 11284932001 |
| Collagenase/Dispase | Sigma | Catalog #: 11097113001 |
| Collagenase P | Sigma | Catalog #: 11213865001 |
| Red Blood Cell Lysing Buffer Hybri-Max | Sigma | Catalog #: R7757-100ML |
| LIVE/DEAD Fixable Aqua Dead Cell Stain Kit | ThermoFisher Scientific | Catalog #: L34966 |
| 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI) | Sigma | Catalog #: 10236276001 |
| Ethylenediaminetetraacetic acid (EDTA) | Sigma | Catalog #: EDS-1KG |
| Sodium azide | Sigma | Catalog #: S2002-500G |
| Methanol | Sigma | Catalog #: 34860-1L-R |
| Formalin solution, neutral buffered, 10% | Sigma | Catalog #: HT501128-4L |
| ProLong Gold Antifade Mountant with DAPI | ThermoFisher Scientific | Catalog #: P36931 |
| ProLong Gold Antifade Mountant | ThermoFisher Scientific | Catalog #: P36930 |
| Critical commercial assays | ||
| Fixation/Permeabilization Solution Kit | BD Bioscience | Catalog #: 554714 |
| Transcription Factor Buffer Set | BD Bioscience | Catalog #: 562574 |
| RNAprotect Cell Reagent | QIAGEN | Catalog #: 76526 |
| RPMI 1640 Medium (Mod.) 1X with L-Glutamine | Fisher Scientific | Catalog #: MT-10043CV |
| DMEM with L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate | Fisher Scientific | Catalog #: MT10013CV |
| IMDM, GlutaMAX | ThermoFisher Scientific | Catalog #: 31980030 |
| L-Glutamine (200 mM) | ThermoFisher Scientific | Catalog #: 25030081 |
| HEPES (1 M) | ThermoFisher Scientific | Catalog #: 15630080 |
| CD8a T Cell Isolation Kit, mouse | Miltenyi Biotec | Catalog #: 130-104-075 |
| B Cell Isolation Kit, mouse | Miltenyi Biotec | Catalog #: 130-090-862 |
| Biotin Tyramide signal amplification | PerkinElmer | Catalog #: NEL700001KT |
| Avidin/Biotin Blocking Kit | Vector Laboratories | Catalog #: SP-2001 |
| Opal 7-Color IHC Kits | Akoya Biosciences | Catalog #: NEL811001KT |
| AR9 Antigen Retrieval Buffer | Akoya Biosciences | Catalog #: AR9001KT |
| RNeasy Micro Kit | QIAGEN | Catalog #: 74004 |
| RNeasy Mini Kit | QIAGEN | Catalog #: 74104 |
| High-Capacity cDNA Reverse Transcription Kit | ThermoFisher Scientific | Catalog #: 4368814 |
| TaqMan Fast Advanced Master Mix | ThermoFisher Scientific | Catalog #: 4444557 |
| Experimental models: Cell lines | ||
| B16-OVA | Hargadon et al. (2006) J. Immunol. 177, 6081 | N/A |
| MC38-OVA | Hargadon et al. (2006) J. Immunol. 177, 6081 | N/A |
| LLC-OVA | Gift of E. Podack (Univ of Miami) | N/A |
| Experimental models: Organisms/strains | ||
| C57BL/6 | Charles River/NCI | Catalog #: 027 |
| Rag1−/− | Jackson Labs | Catalog #: 002216 |
| μMT−/− | Jackson Labs | Catalog #: 002288 |
| LTa−/− | Jackson Labs | Catalog #: 002258 |
| TNFR1/2−/− | Jackson Labs | Catalog #: 003243 |
| Human ubiquitin C Promoter (UBC)-mCherry | Jackson Labs | Catalog #: 017614 |
| CXCR5−/− | Jackson Labs | Catalog #: 006659 |
| Rag2−/−Il2rg−/− | Taconic Labs | Catalog #: 4111-F |
| Oligonucleotides | ||
| CCL21a | ThermoFisher Scientific | Assay ID: Mm03646971_gH |
| CXCL12 | ThermoFisher Scientific | Assay ID: Mm00445552_m1 |
| CXCL13 | ThermoFisher Scientific | Assay ID: Mm00444533_m1 |
| TNFSF13 (APRIL) | ThermoFisher Scientific | Assay ID: Mm03809849_s1 |
| TNFSF13b (BAFF) | ThermoFisher Scientific | Assay ID: Mm01218923_g1 |
| Software and algorithms | ||
| FlowJo v10.7 software | BD Bioscience | https://www.flowjo.com/ |
| ImageJ software | NIH | https://imagej.nih.gov/ij/ |
| Imaris image analysis software | Bitplane | https://imaris.oxinst.com/ |
| Photoshop 2021 | Adobe | https://www.adobe.com/ |
| Vectra 3 Automated Quantitative Pathology Imaging System | Akoya Biosciences | https://www.akoyabio.com/ |
| Phenochart Whole Slide Viewer | Akoya Biosciences | https://www.akoyabio.com/ |
| InForm Tissue Analysis Software | Akoya Biosciences | https://www.akoyabio.com/ |
| HALO Image Analysis Platform Software | IndicaLabs | https://indicalab.com/halo/ |
| QuantStudio 6 Flex Real-Time PCR system | Applied Biosystem | https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/real-time-pcr-instruments/quantstudio-systems/models/quantstudio-6-7-flex.html |
| R version 4 | R Studios | https://www.r-project.org/ |
| Prism version 7.0 | GraphPad | https://www.graphpad.com/ |
| SAS software suite version 9.4 | SAS | https://www.sas.com/en_us/home.html |