Administration of low-dose combination anti-CTLA4, anti-CD137, and anti-OX40 into murine tumor or proximal to the tumor draining lymph node induces systemic tumor regression
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The delivery of immunomodulators directly into the tumor potentially harnesses the existing antigen, tumor-specific infiltrating lymphocytes, and antigen presenting cells. This can confer specificity and generate a potent systemic anti-tumor immune response with lower doses and less toxicity compared to systemic administration, in effect an in situ vaccine. Here, we test this concept using the novel combination of immunomodulators anti-CTLA4, -CD137, and -OX40. The triple combination administered intratumorally at low doses to one tumor of a dual tumor mouse model had dramatic local and systemic anti-tumor efficacy in lymphoma (A20) and solid tumor (MC38) models, consistent with an abscopal effect. The minimal effective dose was 10 μg each. The effect was dependent on CD8 T-cells. Intratumoral administration resulted in superior local and distant tumor control compared to systemic routes, supporting the in situ vaccine concept. In a single tumor A20 model, injection close to the tDLN resulted in similar efficacy as intratumoral and significantly better than targeting a non-tDLN, supporting the role of the tDLN as a viable immunotherapy target in addition to the tumor itself. Distribution studies confirmed expected concentration of antibodies in tumor and tDLN, in keeping with the anti-tumor results. Overall intratumoral or peri-tDLN administration of the novel combination of anti-CTLA4, anti-CD137, and anti-OX40, all agents in the clinic or clinical trials, demonstrates potent systemic anti-tumor effects. This immunotherapeutic combination is promising for future clinical development via both these safe and highly efficacious routes of administration.
KeywordsCombination immunotherapy Intratumoral Tumor draining lymph node CD137 OX40 CTLA4
Enzyme-linked immunosorbent assay
Tumor draining lymph node
Tumor necrosis factor receptor
The authors would like to dedicate this study to the memory of Dr. Holbrook Kohrt, who conceptualized and inspired this work but sadly passed away before its completion. He was a great scientist, physician, and human being. He continues to inspire. Also thanks to Dr. Idit Sagiv-Barfi and Dr. Suparna Dutt for their advice and expertise during the course of the project.
Compliance with ethical standards
Conflict of interest
D. Felsher and J. Hebb received research funding from Alligator Biosciences. P. Ellmark and A. Rosén are employees of Alligator Biosciences. The remaining authors have no conflicts of interest to declare.
This project was funded by NIH grants R01 CA170378, U01 CA188383, and R01 CA184384 (D. W. Felsher) and Alligator Biosciences.
- 1.Hebb J, Kohrt H (2015) Systemic antitumor effects of intratumoral administration of the novel immunotherapeutic combination anti-CTLA4, anti-CD137, and anti-OX40 in mouse models of lymphoma and solid tumor [abstract]. In: American Society of Hematology 57th Annual Meeting; 2015 Dec 5–8; Orlando, FLA. Washington (DC): ASH; 2015. Abstract nr 1552. Blood, vol 126, no 23Google Scholar
- 2.Hebb J, Mosley A, Vences Catalan F, Ellmark P, Norlen P, Felsher D (2016) Intratumoral administration of the immunotherapeutic combination anti-ctla4, anti-cd137 and anti-ox40: comparison to systemic administration, peri-draining lymph node injection, and cellular vaccine in a mouse lymphoma model [abstract]. In: American Society of Hematology 58th Annual Meeting; 2016 Dec 3-6; San Diego, CA. Washington (DC): ASH; 2016. Abstract nr 4172. Blood, vol 128, no 22Google Scholar
- 21.Tirapu I, Mazzolini G, Rodriguez-Calvillo M, Arina A, Palencia B, Gabari I, Melero I (2002) Effective tumor immunotherapy: start the engine, release the brakes, step on the gas pedal,…and get ready to face autoimmunity. Arch Immunol Ther Exp (Warsz) 50(1):13–18Google Scholar
- 35.van Elsas A, Hurwitz AA, Allison JP (1999) Combination immunotherapy of B16 melanoma using anti-cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and granulocyte/macrophage colony-stimulating factor (GM-CSF)-producing vaccines induces rejection of subcutaneous and metastatic tumors accompanied by autoimmune depigmentation. J Exp Med 190(3):355–366CrossRefPubMedPubMedCentralGoogle Scholar