Pancreatic adenocarcinoma is one of the leading causes of cancer-related deaths, and its therapy remains a challenge. Our proposed therapeutic approach is based on the intratumoral injections of mannan-BAM, toll-like receptor ligands, and anti-CD40 antibody (thus termed MBTA therapy), and has shown promising results in the elimination of subcutaneous murine melanoma, pheochromocytoma, colon carcinoma, and smaller pancreatic adenocarcinoma (Panc02). Here, we tested the short- and long-term effects of MBTA therapy in established subcutaneous Panc02 tumors two times larger than in previous study and bilateral Panc02 models as well as the roles of CD4+ and CD8+ T lymphocytes in this therapy. The MBTA therapy resulted in eradication of 67% of Panc02 tumors with the development of long-term memory as evidenced by the rejection of Panc02 cells after subcutaneous and intracranial transplantations. The initial Panc02 tumor elimination is not dependent on the presence of CD4+ T lymphocytes, although these cells seem to be important in long-term survival and resistance against tumor retransplantation. The resistance was revealed to be antigen-specific due to its inability to reject B16-F10 melanoma cells. In the bilateral Panc02 model, MBTA therapy manifested a lower therapeutic response. Despite numerous combinations of MBTA therapy with other therapeutic approaches, our results show that only simultaneous application of MBTA therapy into both tumors has potential for the treatment of the bilateral Panc02 model.

Department of Medical Biology Faculty of Science University of South Bohemia Ceske Budejovice 37005 Czech Republic

Immunoaction LLC Charlotte VT 05445 USA

Section on Medical Neuroendocrinology Eunice Kennedy Shriver National Institute of Child Health and Human Development National Institutes of Health Bethesda MD 20814 USA

Surgical Neurology Branch National Institute of Neurological Disorders and Stroke National Institutes of Health Bethesda MD 20814 USA

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Rawla P, Sunkara T, Gaduputi V. Epidemiology of pancreatic cancer: global trends, etiology and risk factors. World J Oncol. 2019;10(1):10–27. doi: 10.14740/wjon1166. PubMed DOI PMC

Kleeff J, Korc M, Apte M, La Vecchia C, Johnson CD, Biankin AV, Neale RE, Tempero M, Tuveson DA, Hruban RH, Neoptolemos JP. Pancreatic cancer. Nat Rev Dis Primers. 2016;2:16022. doi: 10.1038/nrdp.2016.22. PubMed DOI

Neesse A, Bauer CA, Ohlund D, Lauth M, Buchholz M, Michl P, Tuveson DA, Gress TM. Stromal biology and therapy in pancreatic cancer: ready for clinical translation? Gut. 2019;68(1):159–171. doi: 10.1136/gutjnl-2018-316451. PubMed DOI

Royal RE, Levy C, Turner K, Mathur A, Hughes M, Kammula US, Sherry RM, Topalian SL, Yang JC, Lowy I, Rosenberg SA. Phase 2 trial of single agent Ipilimumab (anti-CTLA-4) for locally advanced or metastatic pancreatic adenocarcinoma. J Immunother. 2010;33(8):828–833. doi: 10.1097/CJI.0b013e3181eec14c. PubMed DOI PMC

Brahmer JR, Tykodi SS, Chow LQ, Hwu WJ, Topalian SL, Hwu P, Drake CG, Camacho LH, Kauh J, Odunsi K, Pitot HC, Hamid O, Bhatia S, Martins R, Eaton K, Chen S, Salay TM, Alaparthy S, Grosso JF, Korman AJ, Parker SM, Agrawal S, Goldberg SM, Pardoll DM, Gupta A, Wigginton JM. Safety and activity of anti-PD-L1 antibody in patients with advanced cancer. N Engl J Med. 2012;366(26):2455–2465. doi: 10.1056/NEJMoa1200694. PubMed DOI PMC

Darvin P, Toor SM, Sasidharan Nair V, Elkord E. Immune checkpoint inhibitors: recent progress and potential biomarkers. Exp Mol Med. 2018;50(12):165. doi: 10.1038/s12276-018-0191-1. PubMed DOI PMC

Hargadon KM, Johnson CE, Williams CJ. Immune checkpoint blockade therapy for cancer: an overview of FDA-approved immune checkpoint inhibitors. Int Immunopharmacol. 2018;62:29–39. doi: 10.1016/j.intimp.2018.06.001. PubMed DOI

Uher O, Caisova V, Hansen P, Kopecky J, Chmelar J, Zhuang Z, Zenka J, Pacak K. Coley’s immunotherapy revived: innate immunity as a link in priming cancer cells for an attack by adaptive immunity. Semin Oncol. 2019 doi: 10.1053/j.seminoncol.2019.10.004. PubMed DOI PMC

Janotova T, Jalovecka M, Auerova M, Svecova I, Bruzlova P, Maierova V, Kumzakova Z, Cunatova S, Vlckova Z, Caisova V, Rozsypalova P, Lukacova K, Vacova N, Wachtlova M, Salat J, Lieskovska J, Kopecky J, Zenka J. The use of anchored agonists of phagocytic receptors for cancer immunotherapy: B16-F10 murine melanoma model. PLoS ONE. 2014;9(1):e85222. doi: 10.1371/journal.pone.0085222. PubMed DOI PMC

Waldmannova E, Caisova V, Faberova J, Svackova P, Kovarova M, Svackova D, Kumzakova Z, Jackova A, Vacova N, Nedbalova P, Horka M, Kopecky J, Zenka J. The use of Zymosan A and bacteria anchored to tumor cells for effective cancer immunotherapy: B16-F10 murine melanoma model. Int Immunopharmacol. 2016;39:295–306. doi: 10.1016/j.intimp.2016.08.004. PubMed DOI

Caisova V, Uher O, Nedbalova P, Jochmanova I, Kvardova K, Masakova K, Krejcova G, Padoukova L, Chmelar J, Kopecky J, Zenka J. Effective cancer immunotherapy based on combination of TLR agonists with stimulation of phagocytosis. Int Immunopharmacol. 2018;59:86–96. doi: 10.1016/j.intimp.2018.03.038. PubMed DOI

Caisova V, Li L, Gupta G, Jochmanova I, Jha A, Uher O, Huynh TT, Miettinen M, Pang Y, Abunimer L, Niu G, Chen X, Ghayee HK, Taieb D, Zhuang Z, Zenka J, Pacak K. The significant reduction or complete eradication of subcutaneous and metastatic lesions in a pheochromocytoma mouse model after immunotherapy using mannan-BAM, TLR ligands, and anti-CD40. Cancers (Basel) 2019 doi: 10.3390/cancers11050654. PubMed DOI PMC

Medina R, Wang HR, Caisova V, Cui J, Indig IH, Uher O, Ye J, Nwankwo A, Sanchez V, Wu TX, Nduom E, Heiss J, Gilbert MR, Terabe M, Ho W, Zenka J, Pacak K, Zhuang ZP. Induction of immune response against metastatic tumors via vaccination of mannan-BAM, TLR ligands, and anti-CD40 antibody (MBTA) Adv Ther Germany. 2020 doi: 10.1002/adtp.202000044. PubMed DOI PMC

Li J, Piao YF, Jiang Z, Chen L, Sun HB. Silencing of signal transducer and activator of transcription 3 expression by RNA interference suppresses growth of human hepatocellular carcinoma in tumor-bearing nude mice. World J Gastroenterol. 2009;15(21):2602–2608. doi: 10.3748/wjg.15.2602. PubMed DOI PMC

Kunk PR, Bauer TW, Slingluff CL, Rahma OE. From bench to bedside a comprehensive review of pancreatic cancer immunotherapy. J Immunother Cancer. 2016;4:14. doi: 10.1186/s40425-016-0119-z. PubMed DOI PMC

Kultti A, Li X, Jiang P, Thompson CB, Frost GI, Shepard HM. Therapeutic targeting of hyaluronan in the tumor stroma. Cancers (Basel) 2012;4(3):873–903. doi: 10.3390/cancers4030873. PubMed DOI PMC

Long KB, Gladney WL, Tooker GM, Graham K, Fraietta JA, Beatty GL. IFNgamma and CCL2 cooperate to redirect tumor-infiltrating monocytes to degrade fibrosis and enhance chemotherapy efficacy in pancreatic carcinoma. Cancer Discov. 2016;6(4):400–413. doi: 10.1158/2159-8290.CD-15-1032. PubMed DOI PMC

Azad A, Yin Lim S, D’Costa Z, Jones K, Diana A, Sansom OJ, Kruger P, Liu S, McKenna WG, Dushek O, Muschel RJ, Fokas E. PD-L1 blockade enhances response of pancreatic ductal adenocarcinoma to radiotherapy. EMBO Mol Med. 2017;9(2):167–180. doi: 10.15252/emmm.201606674. PubMed DOI PMC

Jackaman C, Bundell CS, Kinnear BF, Smith AM, Filion P, van Hagen D, Robinson BW, Nelson DJ. IL-2 intratumoral immunotherapy enhances CD8 + T cells that mediate destruction of tumor cells and tumor-associated vasculature: a novel mechanism for IL-2. J Immunol. 2003;171(10):5051–5063. doi: 10.4049/jimmunol.171.10.5051. PubMed DOI

Haabeth OA, Tveita AA, Fauskanger M, Schjesvold F, Lorvik KB, Hofgaard PO, Omholt H, Munthe LA, Dembic Z, Corthay A, Bogen B. How do CD4(+) T cells detect and eliminate tumor cells that either lack or express MHC class II molecules? Front Immunol. 2014;5:174. doi: 10.3389/fimmu.2014.00174. PubMed DOI PMC

Bogen B, Fauskanger M, Haabeth OA, Tveita A. CD4(+) T cells indirectly kill tumor cells via induction of cytotoxic macrophages in mouse models. Cancer Immunol Immunother. 2019 doi: 10.1007/s00262-019-02374-0. PubMed DOI PMC

Eisel D, Das K, Dickes E, Konig R, Osen W, Eichmuller SB. Cognate interaction with CD4(+) T cells instructs tumor-associated macrophages to acquire M1-like phenotype. Front Immunol. 2019;10:219. doi: 10.3389/fimmu.2019.00219. PubMed DOI PMC

Tang F, Du X, Liu M, Zheng P, Liu Y. Anti-CTLA-4 antibodies in cancer immunotherapy: selective depletion of intratumoral regulatory T cells or checkpoint blockade? Cell Biosci. 2018;8:30. doi: 10.1186/s13578-018-0229-z. PubMed DOI PMC

Park DW, Kim JS, Chin BR, Baek SH. Resveratrol inhibits inflammation induced by heat-killed Listeria monocytogenes. J Med Food. 2012;15(9):788–794. doi: 10.1089/jmf.2012.2194. PubMed DOI PMC

Mathers BW, Harvey HA, Dye CE, Dougherty-Hamod B, Moyer MT. Endoscopic ultrasound-guided ethanol ablation of a large metastatic carcinoid tumor: success with a note of caution. Endosc Int Open. 2014;2(4):E256–E258. doi: 10.1055/s-0034-1377612. PubMed DOI PMC

Anwar MA, Shah M, Kim J, Choi S. Recent clinical trends in Toll-like receptor targeting therapeutics. Med Res Rev. 2019;39(3):1053–1090. doi: 10.1002/med.21553. PubMed DOI PMC

Vonderheide RH. CD40 agonist antibodies in cancer immunotherapy. Annu Rev Med. 2020;71:47–58. doi: 10.1146/annurev-med-062518-045435-045435. PubMed DOI

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