Cancer Immunology, Immunotherapy

, Volume 68, Issue 2, pp 233–245 | Cite as

Development of CDX-1140, an agonist CD40 antibody for cancer immunotherapy

  • Laura A. Vitale
  • Lawrence J. Thomas
  • Li-Zhen He
  • Thomas O’Neill
  • Jenifer Widger
  • Andrea Crocker
  • Karuna Sundarapandiyan
  • James R. Storey
  • Eric M. Forsberg
  • Jeffrey Weidlick
  • April R. Baronas
  • Lauren E. Gergel
  • James M. Boyer
  • Crystal Sisson
  • Joel Goldstein
  • Henry C. MarshJr.
  • Tibor KelerEmail author
Original Article


Limitations of immunotherapy include poorly functioning events early in the immune response cycle, such as efficient antigen presentation and T cell priming. CD40 signaling in dendritic cells leads to upregulation of cell surface costimulatory and MHC molecules and the generation of cytokines, which promotes effective priming of CD8+ effector T cells while minimizing T cell anergy and the generation of regulatory T cells. This naturally occurs through interaction with CD40 ligand (CD40L) expressed on CD4+ T-helper cells. CD40 signaling can also be achieved using specific antibodies, leading to several agonist CD40 antibodies entering clinical development. Our approach to select a CD40 agonist antibody was to define a balanced profile between sufficiently strong immune stimulation and the untoward effects of systemic immune activation. CDX-1140 is a human IgG2 antibody that activates DCs and B cells and drives NFkB stimulation in a CD40-expressing reporter cell line. These activities are Fc-independent and are maintained using an F(ab′)2 fragment of the antibody. CDX-1140 binds outside of the CD40L binding site, and addition of recombinant CD40L greatly enhances DC and B activation by CDX-1140, suggesting that CDX-1140 may act synergistically with naturally expressed CD40L. CDX-1140 also has both direct and immune-mediated anti-tumor activity in xenograft models. CDX-1140 does not promote cytokine production in whole blood assays and has good pharmacodynamic and safety profiles in cynomolgus macaques. These data support the potential of CDX-1140 as part of a cancer therapy regimen, and a phase 1 trial has recently commenced.


CD40 Agonist antibody Immunotherapy Antigen presenting cells 



Association for Assessment and Accreditation of Laboratory Animal Care International


CD40 ligand also referred to as CD154


Chinese hamster ovary


Cysteine-rich domain 1


Cysteine-rich domain 2


Cysteine-rich domain 3


Cysteine-rich domain 4


Cynomolgus macaque


Extracellular domain




Meso Scale Discovery


Maximum tolerated dose


No observable adverse effect level


Recombinant CD40L




TNF receptor associated factors



The authors would like to thank Mallary L. Rocheleau, Michelle E. Grealish, Catherine D. Pilsmaker, Elizabeth Q. Do, Kathleen M. Borrelli, James Testa, Laura Mills-Chen, Collen Patterson and Karla Keler for expert technical assistance.

Author contributions

Laura A. Vitale: study design, methodology, data analysis, and manuscript preparation. Lawrence J. Thomas: study design, methodology, data analysis, and manuscript preparation. Li-Zhen He: study design, methodology, data analysis, and manuscript preparation. Thomas O’Neill: experimental design, methodology, and performance; data analysis. Jenifer Widger: recombinant DNA design, construction and characterization. Andrea Crocker: experimental design, methodology, and performance; data analysis. Karuna Sundarapandiyan: experimental design, methodology, and performance; data analysis. James R. Storey: recombinant DNA design, construction and characterization. Eric M. Forsberg: experimental design, methodology, and performance; data analysis. Jeffrey Weidlick: experimental design, methodology, and performance; data analysis. April R. Baronas: experimental design, methodology, and performance; data analysis. Lauren E. Gergel: experimental design, methodology, and performance; data analysis. James M. Boyer: experimental design, methodology, and performance; data analysis. Crystal Sisson: experimental design, methodology, and performance; data analysis. Joel Goldstein: study design, data analysis, and manuscript preparation. Henry C. Marsh, Jr.: study design, data analysis, and manuscript preparation. Tibor Keler: study design, data analysis, and manuscript preparation.


This work was funded by Celldex Therapeutics, Inc.

Compliance with ethical standards

Conflict of interest

All authors are employees of, and own stock or stock option in Celldex Therapeutics, Inc.

Ethical approval

and ethical standards and animal sources.

Animals were sourced from IACUC-approved commercial sources. Murine xenograft studies (animal source: Taconic Biosciences) were approved by the Celldex IACUC of Hampton, NJ (AUP CDX-002) or the Celldex IACUC of Needham, MA (AUP 08-2017). Animal care followed the Guide for the Care and Use of Laboratory Animals: Eighth Edition (National Research Council. 2011. Washington, DC: The National Academies Press). The pilot primate study (animal source: Charles River Laboratories) was approved by the Charles River Laboratories IACUC of Shrewsbury, MA (AUP 20097548). Those animals were handled according to Guide for the Care and Use of Laboratory Animals: Eighth Edition and AAALAC rules (Association for Assessment and Accreditation of Laboratory Animal Care International). The primate toxicology study (animal source: Kunming Biomed International LTD) was approved by the Citoxlab IACUC of Montreal, QC (AUP 1016–3273). Those animals were handled according to The Canadian Council on Animal Care and AAALAC rules.

Cell line authentication.

Cell lines were sourced directly from vendors that provide authentication. CHO and EJ138 cells were purchased from Millipore-Sigma, HEK-293 cells were purchased from InvivoGen, NFkB luciferase reporter HEK293 stable cell line was purchased from Signosis, and the Ramos and Raji cell lines were purchased from ATCC.

Supplementary material

262_2018_2267_MOESM1_ESM.pdf (202 kb)
Supplementary material 1 (PDF 201 KB)


  1. 1.
    Caux C, Massacrier C, Vanbervliet B, Dubois B, Van Kooten C, Durand I, Banchereau J (1994) Activation of human dendritic cells through CD40 cross-linking. J Exp Med 180:1263–1272CrossRefPubMedGoogle Scholar
  2. 2.
    Clark EA (2014) A Short History of the B-Cell-Associated Surface Molecule CD40. Front Immunol 5:472. CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kiener PA, Moran-Davis P, Rankin BM, Wahl AF, Aruffo A, Hollenbaugh D (1995) Stimulation of CD40 with purified soluble gp39 induces proinflammatory responses in human monocytes. J Immunol 155:4917–4925PubMedGoogle Scholar
  4. 4.
    Henn V, Steinbach S, Buchner K, Presek P, Kroczek RA (2001) The inflammatory action of CD40 ligand (CD154) expressed on activated human platelets is temporally limited by coexpressed CD40. Blood 98:1047–1054CrossRefPubMedGoogle Scholar
  5. 5.
    Yellin MJ, Brett J, Baum D, Matsushima A, Szabolcs M, Stern D, Chess L (1995) Functional interactions of T cells with endothelial cells: the role of CD40L-CD40-mediated signals. J Exp Med 182:1857–1864CrossRefPubMedGoogle Scholar
  6. 6.
    Vonderheide RH, Glennie MJ (2013) Agonistic CD40 antibodies and cancer therapy. Clin Cancer Res 19:1035–1043. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Lederman S, Yellin MJ, Krichevsky A, Belko J, Lee JJ, Chess L (1992) Identification of a novel surface protein on activated CD4 + T cells that induces contact-dependent B cell differentiation (help). J Exp Med 175:1091–1101CrossRefPubMedGoogle Scholar
  8. 8.
    Noelle RJ, Roy M, Shepherd DM, Stamenkovic I, Ledbetter JA, Aruffo A (1992) A 39-kDa protein on activated helper T cells binds CD40 and transduces the signal for cognate activation of B cells. Proc Natl Acad Sci U S A 89:6550–6554CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Reis e Sousa C (2006) Dendritic cells in a mature age. Nat Rev Immunol 6:476–483. CrossRefPubMedGoogle Scholar
  10. 10.
    Yellin MJ, Sinning J, Covey LR et al (1994) T lymphocyte T cell-B cell-activating molecule/CD40-L molecules induce normal B cells or chronic lymphocytic leukemia B cells to express CD80 (B7/BB-1) and enhance their costimulatory activity. J Immunol 153:666–674PubMedGoogle Scholar
  11. 11.
    Beatty GL, Chiorean EG, Fishman MP et al (2011) CD40 agonists alter tumor stroma and show efficacy against pancreatic carcinoma in mice and humans. Science 331:1612–1616. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Wu Y, Wang L, He X, Xu H, Zhou L, Zhao F, Zhang Y (2008) Expression of CD40 and growth-inhibitory activity of CD40 ligand in colon cancer ex vivo. Cell Immunol 253:102–109. CrossRefPubMedGoogle Scholar
  13. 13.
    Zhou Y, He J, Gou LT et al (2012) Expression of CD40 and growth-inhibitory activity of CD40 agonist in ovarian carcinoma cells. Cancer Immunol Immunother 61:1735–1743. CrossRefPubMedGoogle Scholar
  14. 14.
    Wang S, Yang T, Zhu F, Zhu J, Huang Y, Wu L, Chen L, Xu Z (2008) CD40L-mediated inhibition of NF-kappaB in CA46 Burkitt lymphoma cells promotes apoptosis. Leuk Lymphoma 49:1792–1799. CrossRefPubMedGoogle Scholar
  15. 15.
    Advani R, Forero-Torres A, Furman RR, Rosenblatt JD, Younes A, Ren H, Harrop K, Whiting N, Drachman JG (2009) Phase I study of the humanized anti-CD40 monoclonal antibody dacetuzumab in refractory or recurrent non-Hodgkin’s lymphoma. J Clin Oncol 27:4371–4377. CrossRefPubMedGoogle Scholar
  16. 16.
    Bremer E (2013) Targeting of the tumor necrosis factor receptor superfamily for cancer immunotherapy. ISRN Oncol. 2013: 371854.
  17. 17.
    White AL, Chan HT, Roghanian A et al (2011) Interaction with FcgammaRIIB is critical for the agonistic activity of anti-CD40 monoclonal antibody. J Immunol 187:1754–1763. CrossRefPubMedGoogle Scholar
  18. 18.
    Li F, Ravetch JV (2011) Inhibitory Fcgamma receptor engagement drives adjuvant and anti-tumor activities of agonistic CD40 antibodies. Science 333:1030–1034. CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    White AL, Chan HT, French RR et al (2015) Conformation of the human immunoglobulin G2 hinge imparts superagonistic properties to immunostimulatory anticancer antibodies. Cancer Cell 27:138–148. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Beatty GL, Li Y, Long KB (2017) Cancer immunotherapy: activating innate and adaptive immunity through CD40 agonists. Expert Rev Anticancer Ther 17:175–186. CrossRefGoogle Scholar
  21. 21.
    Beatty GL, Torigian DA, Chiorean EG et al (2013) A phase I study of an agonist CD40 monoclonal antibody (CP-870,893) in combination with gemcitabine in patients with advanced pancreatic ductal adenocarcinoma. Clin Cancer Res 19:6286–6295. CrossRefPubMedGoogle Scholar
  22. 22.
    Vonderheide RH, Flaherty KT, Khalil M et al (2007) Clinical activity and immune modulation in cancer patients treated with CP-870,893, a novel CD40 agonist monoclonal antibody. J Clin Oncol 25:876–883. CrossRefPubMedGoogle Scholar
  23. 23.
    Ruter J, Antonia SJ, Burris HA 3rd, Huhn RD, Vonderheide RH (2010) Immune modulation with weekly dosing of an agonist CD40 antibody in a phase I study of patients with advanced solid tumors. Cancer Biol Ther. 10Google Scholar
  24. 24.
    Bajor DL, Xu X, Torigian DA et al (2014) Immune activation and a 9-year ongoing complete remission following CD40 antibody therapy and metastasectomy in a patient with metastatic melanoma. Cancer Immunol Res 2:1051–1058. CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Nowak AK, Cook AM, McDonnell AM et al (2015) A phase 1b clinical trial of the CD40-activating antibody CP-870,893 in combination with cisplatin and pemetrexed in malignant pleural mesothelioma. Ann Oncol 26:2483–2490. CrossRefPubMedGoogle Scholar
  26. 26.
    Morris AE, Remmele RL Jr, Klinke R, Macduff BM, Fanslow WC, Armitage RJ (1999) Incorporation of an isoleucine zipper motif enhances the biological activity of soluble CD40L (CD154). J Biol Chem 274:418–423CrossRefPubMedGoogle Scholar
  27. 27.
    Gladue RP, Paradis T, Cole SH et al (2011) The CD40 agonist antibody CP-870,893 enhances dendritic cell and B-cell activity and promotes anti-tumor efficacy in SCID-hu mice. Cancer Immunol Immunother 60:1009–1017. CrossRefPubMedGoogle Scholar
  28. 28.
    Francisco JA, Donaldson KL, Chace D, Siegall CB, Wahl AF (2000) Agonistic properties and in vivo antitumor activity of the anti-CD40 antibody SGN-14. Cancer Res 60:3225–3231PubMedGoogle Scholar
  29. 29.
    Alexandroff AB, Jackson AM, Paterson T, Haley JL, Ross JA, Longo DL, Murphy WJ, James K, Taub DD (2000) Role for CD40-CD40 ligand interactions in the immune response to solid tumours. Mol Immunol 37:515–526CrossRefPubMedGoogle Scholar
  30. 30.
    Stebbings R, Findlay L, Edwards C et al (2007) “Cytokine storm” in the phase I trial of monoclonal antibody TGN1412: better understanding the causes to improve preclinical testing of immunotherapeutics. J Immunol 179:3325–3331CrossRefPubMedGoogle Scholar
  31. 31.
    Findlay L, Eastwood D, Stebbings R, Sharp G, Mistry Y, Ball C, Hood J, Thorpe R, Poole S (2010) Improved in vitro methods to predict the in vivo toxicity in man of therapeutic monoclonal antibodies including TGN1412. J Immunol Methods 352:1–12CrossRefPubMedGoogle Scholar
  32. 32.
    Sharma P, Allison JP (2015) The future of immune checkpoint therapy. Science 348:56–61. CrossRefPubMedGoogle Scholar
  33. 33.
    Mackey MF, Gunn JR, Ting PP, Kikutani H, Dranoff G, Noelle RJ, Barth RJ Jr (1997) Protective immunity induced by tumor vaccines requires interaction between CD40 and its ligand, CD154. Cancer Res 57:2569–2574PubMedGoogle Scholar
  34. 34.
    French RR, Chan HT, Tutt AL, Glennie MJ (1999) CD40 antibody evokes a cytotoxic T-cell response that eradicates lymphoma and bypasses T-cell help. Nat Med 5:548–553. CrossRefPubMedGoogle Scholar
  35. 35.
    Ngiow SF, Young A, Blake SJ, Hill GR, Yagita H, Teng MW, Korman AJ, Smyth MJ (2016) Agonistic CD40 mAb-Driven IL12 Reverses Resistance to Anti-PD1 in a T-cell-Rich Tumor. Cancer Res 76:6266–6277. CrossRefPubMedGoogle Scholar
  36. 36.
    Grilley-Olson JE, Curti BD, Smith DC et al (2018) SEA-CD40, a non-fucosylated CD40 agonist: Interim results from a phase 1 study in advanced solid tumors. J Clin Oncol 36:3093. CrossRefGoogle Scholar
  37. 37.
    Johnson M, Fakih M, Bendell J, Bajor D, Cristea M, Tremblay T, Trifan O, Vonderheide R (2017) First in human study with the CD40 agonistic monoclonal antibody APX005M in subjects with solid tumors. SITC 2017 Annual Meeting, National Harbor, MDGoogle Scholar
  38. 38.
    Mangsbo SM, Broos S, Fletcher E et al (2015) The human agonistic CD40 antibody ADC-1013 eradicates bladder tumors and generates T-cell-dependent tumor immunity. Clin Cancer Res 21:1115–1126. CrossRefPubMedGoogle Scholar
  39. 39.
    Smulski CR, Beyrath J, Decossas M et al (2013) Cysteine-rich domain 1 of CD40 mediates receptor self-assembly. J Biol Chem 288:10914–10922. CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    An HJ, Kim YJ, Song DH, Park BS, Kim HM, Lee JD, Paik SG, Lee JO, Lee H (2011) Crystallographic and mutational analysis of the CD40-CD154 complex and its implications for receptor activation. J Biol Chem 286:11226–11235. CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Anandasabapathy N, Breton G, Hurley A et al (2015) Efficacy and safety of CDX-301, recombinant human Flt3L, at expanding dendritic cells and hematopoietic stem cells in healthy human volunteers. Bone Marrow Transplant 50:924–930. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Laura A. Vitale
    • 1
  • Lawrence J. Thomas
    • 2
  • Li-Zhen He
    • 1
  • Thomas O’Neill
    • 1
  • Jenifer Widger
    • 1
  • Andrea Crocker
    • 1
  • Karuna Sundarapandiyan
    • 1
  • James R. Storey
    • 2
  • Eric M. Forsberg
    • 2
  • Jeffrey Weidlick
    • 1
  • April R. Baronas
    • 2
  • Lauren E. Gergel
    • 2
  • James M. Boyer
    • 2
  • Crystal Sisson
    • 1
  • Joel Goldstein
    • 1
  • Henry C. MarshJr.
    • 2
  • Tibor Keler
    • 1
    Email author
  1. 1.Celldex Therapeutics, IncHamptonUSA
  2. 2.Celldex Therapeutics, IncNeedhamUSA

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