Journal Article
. 2017 Nov; 8(1):1447.
doi: 10.1038/s41467-017-01572-7.

Intratumoral CD40 activation and checkpoint blockade induces T cell-mediated eradication of melanoma in the brain

Manisha Singh 1 Christina Vianden 1 Mark J Cantwell 2 Zhimin Dai 1 Zhilan Xiao 1 Meenu Sharma 1 Hiep Khong 1 Ashvin R Jaiswal 3 Faisal Faak 1 Yared Hailemichael 1 L M E Janssen 1 Uddalak Bharadwaj 4 Michael A Curran 3 Adi Diab 1 Roland L Bassett 5 David J Tweardy 4 Patrick Hwu 1 Willem W Overwijk 6 
  • PMID: 29129918
  •     34 References
  •     37 citations


CD40 agonists bind the CD40 molecule on antigen-presenting cells and activate them to prime tumor-specific CD8+ T cell responses. Here, we study the antitumor activity and mechanism of action of a nonreplicating adenovirus encoding a chimeric, membrane-bound CD40 ligand (ISF35). Intratumoral administration of ISF35 in subcutaneous B16 melanomas generates tumor-specific, CD8+ T cells that express PD-1 and suppress tumor growth. Combination therapy of ISF35 with systemic anti-PD-1 generates greater antitumor activity than each respective monotherapy. Triple combination of ISF35, anti-PD-1, and anti-CTLA-4 results in complete eradication of injected and noninjected subcutaneous tumors, as well as melanoma tumors in the brain. Therapeutic efficacy is associated with increases in the systemic level of tumor-specific CD8+ T cells, and an increased ratio of intratumoral CD8+ T cells to CD4+ Tregs. These results provide a proof of concept of systemic antitumor activity after intratumoral CD40 triggering with ISF35 in combination with checkpoint blockade for multifocal cancer, including the brain.

Effective innate and adaptive antimelanoma immunity through localized TLR7/8 activation.
Manisha Singh, Hiep Khong, +6 authors, Willem W Overwijk.
J Immunol, 2014 Sep 26; 193(9). PMID: 25252955    Free PMC article.
Localized oncolytic virotherapy overcomes systemic tumor resistance to immune checkpoint blockade immunotherapy.
Dmitriy Zamarin, Rikke B Holmgaard, +6 authors, James P Allison.
Sci Transl Med, 2014 Mar 07; 6(226). PMID: 24598590    Free PMC article.
Highly Cited.
Conversion of tumor-specific CD4+ T-cell tolerance to T-cell priming through in vivo ligation of CD40.
E M Sotomayor, I Borrello, +6 authors, H I Levitsky.
Nat Med, 1999 Jul 08; 5(7). PMID: 10395323
Highly Cited.
CD40 activation in epithelial ovarian carcinoma cells modulates growth, apoptosis, and cytokine secretion.
N J Gallagher, A G Eliopoulos, +3 authors, L S Young.
Mol Pathol, 2002 Apr 16; 55(2). PMID: 11950960    Free PMC article.
In this issue: harnessing immunity against difficult targets, through the CD40-CD40L pathway.
Richard S Kornbluth, Adrian Bot.
Int Rev Immunol, 2012 Jul 19; 31(4). PMID: 22804569
Clinical activity and immune modulation in cancer patients treated with CP-870,893, a novel CD40 agonist monoclonal antibody.
Robert H Vonderheide, Keith T Flaherty, +12 authors, Scott J Antonia.
J Clin Oncol, 2007 Mar 01; 25(7). PMID: 17327609
Highly Cited.
Recent developments on immunotherapy for brain cancer.
Derek A Wainwright, Pragati Nigam, +2 authors, Maciej S Lesniak.
Expert Opin Emerg Drugs, 2012 Apr 27; 17(2). PMID: 22533851    Free PMC article.
Intratumoral injection of Ad-ISF35 (Chimeric CD154) breaks tolerance and induces lymphoma tumor regression.
Mauricio Urquiza, Johanna Melo-Cardenas, +2 authors, Januario E Castro.
Hum Gene Ther, 2014 Nov 11; 26(1). PMID: 25382101    Free PMC article.
Evolving treatment options for melanoma brain metastases.
Thankamma Ajithkumar, Christine Parkinson, +2 authors, Sarah Jefferies.
Lancet Oncol, 2015 Oct 05; 16(13). PMID: 26433822
Nivolumab and ipilimumab versus ipilimumab in untreated melanoma.
Michael A Postow, Jason Chesney, +18 authors, F Stephen Hodi.
N Engl J Med, 2015 Apr 22; 372(21). PMID: 25891304    Free PMC article.
Highly Cited.
Cancer-fighting viruses win approval.
Heidi Ledford.
Nature, 2015 Oct 30; 526(7575). PMID: 26511559
Talimogene laherparepvec (T-VEC) for the treatment of advanced melanoma.
Douglas B Johnson, Igor Puzanov, Mark C Kelley.
Immunotherapy, 2015 Jun 23; 7(6). PMID: 26098919    Free PMC article.
Cancer immunotherapy: moving beyond current vaccines.
Steven A Rosenberg, James C Yang, Nicholas P Restifo.
Nat Med, 2004 Sep 02; 10(9). PMID: 15340416    Free PMC article.
Highly Cited.
Vaccination with a recombinant vaccinia virus encoding a "self" antigen induces autoimmune vitiligo and tumor cell destruction in mice: requirement for CD4(+) T lymphocytes.
W W Overwijk, D S Lee, +7 authors, N P Restifo.
Proc Natl Acad Sci U S A, 1999 Mar 17; 96(6). PMID: 10077623    Free PMC article.
Braf(V600E) cooperates with Pten loss to induce metastatic melanoma.
David Dankort, David P Curley, +7 authors, Marcus Bosenberg.
Nat Genet, 2009 Mar 14; 41(5). PMID: 19282848    Free PMC article.
Highly Cited.
ICP34.5 deleted herpes simplex virus with enhanced oncolytic, immune stimulating, and anti-tumour properties.
B L Liu, M Robinson, +9 authors, R S Coffin.
Gene Ther, 2003 Feb 22; 10(4). PMID: 12595888
Highly Cited.
Clinical Management of Multiple Melanoma Brain Metastases: A Systematic Review.
Sharad Goyal, Ann W Silk, +4 authors, Howard L Kaufman.
JAMA Oncol, 2015 Jul 17; 1(5). PMID: 26181286    Free PMC article.
Systematic Review.
Agonistic antibody to CD40 boosts the antitumor activity of adoptively transferred T cells in vivo.
Chengwen Liu, Carol M Lewis, +8 authors, Patrick Hwu.
J Immunother, 2012 Mar 17; 35(3). PMID: 22421945    Free PMC article.
CD4+CD25+ regulatory T cells in tumor immunity.
Xin Chen, Yong Du, +3 authors, ZhiMing Huang.
Int Immunopharmacol, 2016 Mar 20; 34. PMID: 26994448
The anatomical and cellular basis of immune surveillance in the central nervous system.
Richard M Ransohoff, Britta Engelhardt.
Nat Rev Immunol, 2012 Aug 21; 12(9). PMID: 22903150
Highly Cited. Review.
Phase II clinical trial of a granulocyte-macrophage colony-stimulating factor-encoding, second-generation oncolytic herpesvirus in patients with unresectable metastatic melanoma.
Neil N Senzer, Howard L Kaufman, +12 authors, John J Nemunaitis.
J Clin Oncol, 2009 Nov 04; 27(34). PMID: 19884534
Highly Cited.
The pharmacological impact of ATP-binding cassette drug transporters on vemurafenib-based therapy.
Chung-Pu Wu, Suresh V Ambudkar.
Acta Pharm Sin B, 2014 Apr 01; 4(2). PMID: 26579371    Free PMC article.
Melanoma brain metastasis: overview of current management and emerging targeted therapies.
Ekokobe Fonkem, Erik J Uhlmann, +4 authors, Eric T Wong.
Expert Rev Neurother, 2012 Oct 23; 12(10). PMID: 23082737
CD40 expression in bladder cancer.
P W Cooke, N D James, +3 authors, L S Young.
J Pathol, 1999 Jul 09; 188(1). PMID: 10398138
CD40 Stimulation Obviates Innate Sensors and Drives T Cell Immunity in Cancer.
Katelyn T Byrne, Robert H Vonderheide.
Cell Rep, 2016 Jun 14; 15(12). PMID: 27292635    Free PMC article.
Highly Cited.
Immunization against endogenous retroviral tumor-associated antigens.
M H Kershaw, C Hsu, +8 authors, P Hwu.
Cancer Res, 2001 Nov 03; 61(21). PMID: 11691813    Free PMC article.
Intratumoral delivery of CD154 homolog (Ad-ISF35) induces tumor regression: analysis of vector biodistribution, persistence and gene expression.
J Melo-Cardenas, M Urquiza, T J Kipps, J E Castro.
Cancer Gene Ther, 2012 Mar 10; 19(5). PMID: 22402624    Free PMC article.
CD40-induced aggregation of MHC class II and CD80 on the cell surface leads to an early enhancement in antigen presentation.
Abigail Clatza, Laura C Bonifaz, Dario A A Vignali, José Moreno.
J Immunol, 2003 Dec 10; 171(12). PMID: 14662847
Gene immunotherapy of chronic lymphocytic leukemia: a phase I study of intranodally injected adenovirus expressing a chimeric CD154 molecule.
Januario E Castro, Johanna Melo-Cardenas, +3 authors, Thomas J Kipps.
Cancer Res, 2012 Apr 17; 72(12). PMID: 22505652    Free PMC article.
CD40 expression by human melanocytic lesions and melanoma cell lines and direct CD40 targeting with the therapeutic anti-CD40 antibody CP-870,893.
Anusha Kalbasi, Ester Fonsatti, +9 authors, Antoni Ribas.
J Immunother, 2010 Sep 16; 33(8). PMID: 20842056
Agonistic CD40 antibodies and cancer therapy.
Robert H Vonderheide, Martin J Glennie.
Clin Cancer Res, 2013 Mar 06; 19(5). PMID: 23460534    Free PMC article.
Highly Cited.
T-bet and eomesodermin are required for T cell-mediated antitumor immune responses.
Yibei Zhu, Songguang Ju, +6 authors, Binfeng Lu.
J Immunol, 2010 Aug 18; 185(6). PMID: 20713880
Fc-dependent depletion of tumor-infiltrating regulatory T cells co-defines the efficacy of anti-CTLA-4 therapy against melanoma.
Tyler R Simpson, Fubin Li, +11 authors, Sergio A Quezada.
J Exp Med, 2013 Jul 31; 210(9). PMID: 23897981    Free PMC article.
Highly Cited.
Depleting tumor-specific Tregs at a single site eradicates disseminated tumors.
Aurélien Marabelle, Holbrook Kohrt, +13 authors, Ronald Levy.
J Clin Invest, 2013 Jun 04; 123(6). PMID: 23728179    Free PMC article.
Highly Cited.
Oncolytic virus and PD-1/PD-L1 blockade combination therapy.
Chun-Yu Chen, Brian Hutzen, Mary F Wedekind, Timothy P Cripe.
Oncolytic Virother, 2018 Aug 15; 7. PMID: 30105219    Free PMC article.
Treatment with mRNA coding for the necroptosis mediator MLKL induces antitumor immunity directed against neo-epitopes.
Lien Van Hoecke, Sandra Van Lint, +6 authors, Xavier Saelens.
Nat Commun, 2018 Aug 26; 9(1). PMID: 30143632    Free PMC article.
Synergistic cancer immunotherapy combines MVA-CD40L induced innate and adaptive immunity with tumor targeting antibodies.
José Medina-Echeverz, Maria Hinterberger, +11 authors, Henning Lauterbach.
Nat Commun, 2019 Nov 07; 10(1). PMID: 31695037    Free PMC article.
Modeling multi-needle injection into solid tumor.
Vladimir Subbotin, Gennady Fiksel.
Am J Cancer Res, 2019 Nov 14; 9(10). PMID: 31720083    Free PMC article.
Immune System Evasion as Hallmark of Melanoma Progression: The Role of Dendritic Cells.
Marco Tucci, Anna Passarelli, +4 authors, Francesco Silvestris.
Front Oncol, 2019 Nov 22; 9. PMID: 31750245    Free PMC article.
Melittin-lipid nanoparticles target to lymph nodes and elicit a systemic anti-tumor immune response.
Xiang Yu, Yanfeng Dai, +5 authors, Zhihong Zhang.
Nat Commun, 2020 Mar 01; 11(1). PMID: 32111828    Free PMC article.
A CD40 Agonist and PD-1 Antagonist Antibody Reprogram the Microenvironment of Nonimmunogenic Tumors to Allow T-cell-Mediated Anticancer Activity.
Hayley S Ma, Bibhav Poudel, +9 authors, Elizabeth M Jaffee.
Cancer Immunol Res, 2019 Jan 16; 7(3). PMID: 30642833    Free PMC article.
Concepts for agonistic targeting of CD40 in immuno-oncology.
David M Richards, Julian P Sefrin, +2 authors, Christian Merz.
Hum Vaccin Immunother, 2019 Aug 14; 16(2). PMID: 31403344    Free PMC article.
A Novel Signature of 23 Immunity-Related Gene Pairs Is Prognostic of Cutaneous Melanoma.
Ya-Nan Xue, Yi-Nan Xue, +3 authors, Wei-Qiang Tan.
Front Immunol, 2020 Nov 17; 11. PMID: 33193373    Free PMC article.
Correlation of tumor-infiltrating immune cells of melanoma with overall survival by immunogenomic analysis.
Lili Huang, Hong Chen, +3 authors, Qing Xu.
Cancer Med, 2020 Sep 16; 9(22). PMID: 32931642    Free PMC article.
New emerging targets in cancer immunotherapy: the role of Cluster of Differentiation 40 (CD40/TNFR5).
Manuel Piechutta, Anna Sophie Berghoff.
ESMO Open, 2019 Jul 06; 4(Suppl 3). PMID: 31275618    Free PMC article.
Ad-CD40L mobilizes CD4 T cells for the treatment of brainstem tumors.
Phonphimon Wongthida, Matthew R Schuelke, +10 authors, Richard G Vile.
Neuro Oncol, 2020 May 28; 22(12). PMID: 32459315    Free PMC article.
Emerging immunotherapies for metastasis.
Sarah C Edwards, Wilma H M Hoevenaar, Seth B Coffelt.
Br J Cancer, 2020 Dec 03; 124(1). PMID: 33262520    Free PMC article.
Boiling histotripsy and in-situ CD40 stimulation improve the checkpoint blockade therapy of poorly immunogenic tumors.
Mohit Pratap Singh, Sri Nandhini Sethuraman, +2 authors, Ashish Ranjan.
Theranostics, 2021 Jan 05; 11(2). PMID: 33391491    Free PMC article.
M1-like TAMs are required for the efficacy of PD-L1/PD-1 blockades in gastric cancer.
Rui Zhao, Qianyi Wan, +11 authors, Xiaoting Wu.
Oncoimmunology, 2021 Jan 19; 10(1). PMID: 33457080    Free PMC article.
A Hepatitis B Virus-Derived Peptide Exerts an Anticancer Effect via TNF/iNOS-producing Dendritic Cells in Tumor-Bearing Mouse Model.
Soo-Bin Yang, Mi-Hyun Lee, +2 authors, Bum-Joon Kim.
Cancers (Basel), 2021 Jan 28; 13(3). PMID: 33499256    Free PMC article.
Molecular basis and therapeutic implications of CD40/CD40L immune checkpoint.
TingTing Tang, Xiang Cheng, +3 authors, Hong Wang.
Pharmacol Ther, 2020 Oct 23; 219. PMID: 33091428    Free PMC article.
Varied functions of immune checkpoints during cancer metastasis.
Ali Safarzadeh, Mohsen Alizadeh, +7 authors, Nicola Silvestris.
Cancer Immunol Immunother, 2020 Sep 10; 70(3). PMID: 32902664    Free PMC article.
IL-1α Mediates Innate and Acquired Resistance to Immunotherapy in Melanoma.
Shubhra Singh, Zhilan Xiao, +10 authors, Manisha Singh.
J Immunol, 2021 Mar 17; 206(8). PMID: 33722878    Free PMC article.
Novel induction of CD40 expression by tumor cells with RAS/RAF/PI3K pathway inhibition augments response to checkpoint blockade.
Chi Yan, Nabil Saleh, +9 authors, Ann Richmond.
Mol Cancer, 2021 Jun 08; 20(1). PMID: 34092233    Free PMC article.
Co-Stimulatory versus Cell Death Aspects of Agonistic CD40 Monoclonal Antibody Selicrelumab in Chronic Lymphocytic Leukemia.
Raquel Delgado, Karoline Kielbassa, +5 authors, Eric Eldering.
Cancers (Basel), 2021 Jul 03; 13(12). PMID: 34205588    Free PMC article.
Pyroptosis-Related Gene Signatures Can Robustly Diagnose Skin Cutaneous Melanoma and Predict the Prognosis.
Anji Ju, Jiaze Tang, +2 authors, Yongzhang Luo.
Front Oncol, 2021 Jul 31; 11. PMID: 34327145    Free PMC article.
Dendritic cell targeting with Fc-enhanced CD40 antibody agonists induces durable antitumor immunity in humanized mouse models of bladder cancer.
Christopher S Garris, Jeffrey L Wong, Jeffrey V Ravetch, David A Knorr.
Sci Transl Med, 2021 May 21; 13(594). PMID: 34011627    Free PMC article.
Adenoviral CD40 Ligand Immunotherapy in 32 Canine Malignant Melanomas-Long-Term Follow Up.
Sara Saellstrom, Arian Sadeghi, +7 authors, Henrik Ronnberg.
Front Vet Sci, 2021 Aug 10; 8. PMID: 34368282    Free PMC article.
Next-generation immunotherapies for brain metastatic cancers.
María López Vázquez, Wanlu Du, +2 authors, Khalid Shah.
Trends Cancer, 2021 Mar 17; 7(9). PMID: 33722479    Free PMC article.
TNF blockade uncouples toxicity from antitumor efficacy induced with CD40 chemoimmunotherapy.
Meredith L Stone, Jesse Lee, +11 authors, Gregory L Beatty.
JCI Insight, 2021 Jun 09; 6(14). PMID: 34101617    Free PMC article.
Pre-clinical evaluation of immunoPET imaging using agonist CD40 monoclonal antibody in pancreatic tumor-bearing mice.
Sadaf Aghevlian, Bo Wu, +4 authors, Katherine W Ferrara.
Nucl Med Biol, 2021 May 08; 98-99. PMID: 33962357    Free PMC article.
Ex vivo expanded allogeneic natural killer cells have potent cytolytic activity against cancer cells through different receptor-ligand interactions.
Daun Jung, Young Seok Baek, +8 authors, Hee Jung An.
J Exp Clin Cancer Res, 2021 Oct 24; 40(1). PMID: 34686187    Free PMC article.
Kickstarting Immunity in Cold Tumours: Localised Tumour Therapy Combinations With Immune Checkpoint Blockade.
Elizabeth Appleton, Jehanne Hassan, +7 authors, Erik Wennerberg.
Front Immunol, 2021 Nov 05; 12. PMID: 34733287    Free PMC article.
Simultaneous Inhibition of PD-1 and Stimulation of CD40 Signaling Pathways by Anti-PD-L1/CD40L Bispecific Fusion Protein Synergistically Activate Target and Effector Cells.
Madhu S Pandey, Chunlei Wang, Scott Umlauf, Shihua Lin.
Int J Mol Sci, 2021 Nov 14; 22(21). PMID: 34768776    Free PMC article.
Myeloid-Derived Suppressive Cell Expansion Promotes Melanoma Growth and Autoimmunity by Inhibiting CD40/IL27 Regulation in Macrophages.
Julio C Valencia, Rebecca A Erwin-Cohen, +18 authors, Howard A Young.
Cancer Res, 2021 Oct 14; 81(23). PMID: 34642183    Free PMC article.
Directing T-Cell Immune Responses for Cancer Vaccination and Immunotherapy.
Peter Lawrence Smith, Katarzyna Piadel, Angus George Dalgleish.
Vaccines (Basel), 2021 Dec 29; 9(12). PMID: 34960140    Free PMC article.
Combination therapy with immune checkpoint inhibitors (ICIs); a new frontier.
Somayeh Vafaei, Angelina O Zekiy, +4 authors, Majid Zamani.
Cancer Cell Int, 2022 Jan 05; 22(1). PMID: 34980128    Free PMC article.
CD40 stimulation as a molecular adjuvant for cancer vaccines and other immunotherapies.
Timothy N J Bullock.
Cell Mol Immunol, 2021 Jul 21; 19(1). PMID: 34282297    Free PMC article.
Mechanisms of immune activation and regulation: lessons from melanoma.
Shelly Kalaora, Adi Nagler, Jennifer A Wargo, Yardena Samuels.
Nat Rev Cancer, 2022 Feb 03; 22(4). PMID: 35105962
Immune priming using DC- and T cell-targeting gene therapy sensitizes both treated and distant B16 tumors to checkpoint inhibition.
Jessica Wenthe, Sedigheh Naseri, +6 authors, Angelica Loskog.
Mol Ther Oncolytics, 2022 Feb 11; 24. PMID: 35141399    Free PMC article.
Tumor immunotherapies by immune checkpoint inhibitors (ICIs); the pros and cons.
Adel Naimi, Rebar N Mohammed, +11 authors, Ehsan Razeghian.
Cell Commun Signal, 2022 Apr 09; 20(1). PMID: 35392976    Free PMC article.