The natural adjuvant properties of bacterial ghosts (BGs) lie within the presence of intact pathogen-associated molecular patterns on their surface. BGs can improve the direct delivery, natural processing and presentation of target antigens within dendritic cells (DCs). Moreover, sensitization of human DCs by cancer cell lysate (oncolysate)-loaded BGs in the presence of IFN-α and GM-CSF enhanced DC maturation as indicated by an increased expression of maturation markers and co-stimulatory molecules, higher production of IL-12p70 and stimulation of significantly increased proliferation of both autologous CD4+ and CD8+ T cells compared to DCs matured in the presence of purified lipopolysaccharide. The induced T cells efficiently recognized oncolysate-derived tumor-associated antigens expressed by cancer cells used for the production of oncolysate. Our optimized one-step simultaneous antigen delivery and DC maturation-inducing method emerges as a promising tool for the development and implementation of next-generation cellular cancer immunotherapies.

Advanced Cell Immunotherapy Unit Department of Pharmacology Medical Faculty Masaryk University Brno Czech Republic

BIRD C GmbH Dr Bohrgasse 2 8 1030 Vienna Austria

Cellthera s r o Brno Czech Republic

Department of Immunology National Cancer Institute Vilnius Lithuania

Department of Pediatrics The University Hospital Brno Brno Czech Republic

Department of Pharmacology and Immunotherapy Veterinary Research Institute Brno Czech Republic

Institute of Mathematics and Physics Faculty of Mechanical Engineering Slovak University of Technology Bratislava Slovak Republic

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Stewart BW, Wild CP (eds) (2014) World cancer report 2014. International agency for research on cancer, Lyon, France PubMed

Chen G, Emens LA. Chemoimmunotherapy: reengineering tumor immunity. Cancer Immunol Immunother. 2013;62(2):203–216. doi: 10.1007/s00262-012-1388-0. PubMed DOI PMC

Vanneman M, Dranoff G. Combining immunotherapy and targeted therapies in cancer treatment. Nat Rev Cancer. 2012;12(4):237–251. doi: 10.1038/nrc3237. PubMed DOI PMC

Bhatia A, Kumar Y. Cellular and molecular mechanisms in cancer immune escape: a comprehensive review. Expert Rev Clin Immunol. 2014;10(1):41–62. doi: 10.1586/1744666X.2014.865519. PubMed DOI

Zitvogel L, Galluzzi L, Smyth MJ, Kroemer G. Mechanism of action of conventional and targeted anticancer therapies: reinstating immunosurveillance. Immunity. 2013;39(1):74–88. doi: 10.1016/j.immuni.2013.06.014. PubMed DOI

Nguyen T, Urban J, Kalinski P. Therapeutic cancer vaccines and combination immunotherapies involving vaccination. Immunotargets Ther. 2014;3:135–150. PubMed PMC

Schijns V, Tartour E, Michalek J, Stathopoulos A, Dobrovolskiene NT, Strioga MM. Immune adjuvants as critical guides directing immunity triggered by therapeutic cancer vaccines. Cytotherapy. 2014;16(4):427–439. doi: 10.1016/j.jcyt.2013.09.008. PubMed DOI

Palucka K, Banchereau J. Dendritic-cell-based therapeutic cancer vaccines. Immunity. 2013;39(1):38–48. doi: 10.1016/j.immuni.2013.07.004. PubMed DOI PMC

Strioga MM, Felzmann T, Powell DJ, Jr, Ostapenko V, Dobrovolskiene NT, Matuskova M, Michalek J, Schijns VE. Therapeutic dendritic cell-based cancer vaccines: the state of the art. Crit Rev Immunol. 2013;33(6):489–547. doi: 10.1615/CritRevImmunol.2013008033. PubMed DOI

Kantoff PW, Higano CS, Shore ND, Berger ER, Small EJ, Penson DF, Redfern CH, Ferrari AC, Dreicer R, Sims RB, Xu Y, Frohlich MW, Schellhammer PF. Sipuleucel-T immunotherapy for castration-resistant prostate cancer. N Engl J Med. 2010;363(5):411–422. doi: 10.1056/NEJMoa1001294. PubMed DOI

Butterfield LH. Dendritic cells in cancer immunotherapy clinical trials: are we making progress? Front Immunol. 2013;4:454. doi: 10.3389/fimmu.2013.00454. PubMed DOI PMC

Anguille S, Smits EL, Lion E, van Tendeloo VF, Berneman ZN. Clinical use of dendritic cells for cancer therapy. Lancet Oncol. 2014;15(7):e257–e267. doi: 10.1016/S1470-2045(13)70585-0. PubMed DOI

Datta J, Terhune JH, Lowenfeld L, Cintolo JA, Xu S, Roses RE, Czerniecki BJ. Optimizing dendritic cell-based approaches for cancer immunotherapy. Yale J Biol Med. 2014;87(4):491–518. PubMed PMC

Mailliard RB, Wankowicz-Kalinska A, Cai Q, Wesa A, Hilkens CM, Kapsenberg ML, Kirkwood JM, Storkus WJ, Kalinski P. Alpha-type-1 polarized dendritic cells: a novel immunization tool with optimized CTL-inducing activity. Cancer Res. 2004;64(17):5934–5937. doi: 10.1158/0008-5472.CAN-04-1261. PubMed DOI

Lee MKt, Xu S, Fitzpatrick EH, Sharma A, Graves HL, Czerniecki BJ. Inhibition of CD4+ CD25+ regulatory T cell function and conversion into Th1-like effectors by a Toll-like receptor-activated dendritic cell vaccine. PLoS ONE. 2013;8(11):e74698. doi: 10.1371/journal.pone.0074698. PubMed DOI PMC

Vopenkova K, Mollova K, Buresova I, Michalek J. Complex evaluation of human monocyte-derived dendritic cells for cancer immunotherapy. J Cell Mol Med. 2012;16(11):2827–2837. doi: 10.1111/j.1582-4934.2012.01614.x. PubMed DOI PMC

Sundarasetty BS, Chan L, Darling D, Giunti G, Farzaneh F, Schenck F, Naundorf S, Kuehlcke K, Ruggiero E, Schmidt M, von Kalle C, Rothe M, Hoon DS, Gerasch L, Figueiredo C, Koehl U, Blasczyk R, Gutzmer R, Stripecke R. Lentivirus-induced ‘Smart’ dendritic cells: pharmacodynamics and GMP-compliant production for immunotherapy against TRP2-positive melanoma. Gene Ther. 2015;22(9):707–720. doi: 10.1038/gt.2015.43. PubMed DOI PMC

Powell KL, Stephens AS, Ralph SJ. Development of a potent melanoma vaccine capable of stimulating CD8(+) T-cells independently of dendritic cells in a mouse model. Cancer Immunol Immunother. 2015;64(7):861–872. doi: 10.1007/s00262-015-1695-3. PubMed DOI PMC

Muhammad A, Champeimont J, Mayr UB, Lubitz W, Kudela P. Bacterial ghosts as carriers of protein subunit and DNA-encoded antigens for vaccine applications. Expert Rev Vaccines. 2012;11(1):97–116. doi: 10.1586/erv.11.149. PubMed DOI

Ebensen T, Paukner S, Link C, Kudela P, de Domenico C, Lubitz W, Guzman CA. Bacterial ghosts are an efficient delivery system for DNA vaccines. J Immunol. 2004;172(11):6858–6865. doi: 10.4049/jimmunol.172.11.6858. PubMed DOI

Kudela P, Paukner S, Mayr UB, Cholujova D, Schwarczova Z, Sedlak J, Bizik J, Lubitz W. Bacterial ghosts as novel efficient targeting vehicles for DNA delivery to the human monocyte-derived dendritic cells. J Immunother. 2005;28(2):136–143. doi: 10.1097/01.cji.0000154246.89630.6f. PubMed DOI

Kudela P, Schwarczova Z, Sedlak J, Bizik J. Conditioned medium from HeLa cells enhances motility of human monocyte-derived dendritic cells but abrogates their maturation and endocytic activity. Neoplasma. 2001;48(5):382–388. PubMed

Ma Y, Shurin GV, Peiyuan Z, Shurin MR. Dendritic cells in the cancer microenvironment. J Cancer. 2013;4(1):36–44. doi: 10.7150/jca.5046. PubMed DOI PMC

Langemann T, Koller VJ, Muhammad A, Kudela P, Mayr UB, Lubitz W. The bacterial ghost platform system: production and applications. Bioeng Bugs. 2010;1(5):326–336. doi: 10.4161/bbug.1.5.12540. PubMed DOI PMC

Jonuleit H, Kuhn U, Muller G, Steinbrink K, Paragnik L, Schmitt E, Knop J, Enk AH. Pro-inflammatory cytokines and prostaglandins induce maturation of potent immunostimulatory dendritic cells under fetal calf serum-free conditions. Eur J Immunol. 1997;27(12):3135–3142. doi: 10.1002/eji.1830271209. PubMed DOI

Kalinski P. Regulation of immune responses by prostaglandin E2. J Immunol. 2012;188(1):21–28. doi: 10.4049/jimmunol.1101029. PubMed DOI PMC

Witte A, Wanner G, Sulzner M, Lubitz W. Dynamics of PhiX174 protein E-mediated lysis of Escherichia coli . Arch Microbiol. 1992;157(4):381–388. doi: 10.1007/BF00248685. PubMed DOI

Eko FO, Mania-Pramanik J, Pais R, Pan Q, Okenu DM, Johnson A, Ibegbu C, He C, He Q, Russell R, Black CM, Igietseme JU. Vibrio cholerae ghosts (VCG) exert immunomodulatory effect on dendritic cells for enhanced antigen presentation and induction of protective immunity. BMC Immunol. 2014;15(1):584. doi: 10.1186/s12865-014-0056-x. PubMed DOI PMC

Kudela P, Paukner S, Mayr UB, Cholujova D, Kohl G, Schwarczova Z, Bizik J, Sedlak J, Lubitz W. Effective gene transfer to melanoma cells using bacterial ghosts. Cancer Lett. 2008;262(1):54–63. doi: 10.1016/j.canlet.2007.11.031. PubMed DOI

Paukner S, Kudela P, Kohl G, Schlapp T, Friedrichs S, Lubitz W. DNA-loaded bacterial ghosts efficiently mediate reporter gene transfer and expression in macrophages. Mol Ther. 2005;11(2):215–223. doi: 10.1016/j.ymthe.2004.09.024. PubMed DOI

de Rosa F, Ridolfi L, Ridolfi R, Gentili G, Valmorri L, Nanni O, Petrini M, Fiammenghi L, Granato AM, Ancarani V, Pancisi E, Soldati V, Cassan S, Riccobon A, Parisi E, Romeo A, Turci L, Guidoboni M. Vaccination with autologous dendritic cells loaded with autologous tumor lysate or homogenate combined with immunomodulating radiotherapy and/or preleukapheresis IFN-alpha in patients with metastatic melanoma: a randomised “proof-of-principle” phase II study. J Transl Med. 2014;12:209. doi: 10.1186/1479-5876-12-209. PubMed DOI PMC

Eyrich M, Schreiber SC, Rachor J, Krauss J, Pauwels F, Hain J, Wolfl M, Lutz MB, de Vleeschouwer S, Schlegel PG, Van Gool SW. Development and validation of a fully GMP-compliant production process of autologous, tumor-lysate-pulsed dendritic cells. Cytotherapy. 2014;16(7):946–964. doi: 10.1016/j.jcyt.2014.02.017. PubMed DOI

Gonzalez FE, Ortiz C, Reyes M, Dutzan N, Patel V, Pereda C, Gleisner MA, Lopez MN, Gutkind JS, Salazar-Onfray F. Melanoma cell lysate induces CCR7 expression and in vivo migration to draining lymph nodes of therapeutic human dendritic cells. Immunology. 2014;142(3):396–405. doi: 10.1111/imm.12264. PubMed DOI PMC

Chiang CL, Kandalaft LE, Tanyi J, Hagemann AR, Motz GT, Svoronos N, Montone K, Mantia-Smaldone GM, Smith L, Nisenbaum HL, Levine BL, Kalos M, Czerniecki BJ, Torigian DA, Powell DJ, Jr, Mick R, Coukos G. A dendritic cell vaccine pulsed with autologous hypochlorous acid-oxidized ovarian cancer lysate primes effective broad antitumor immunity: from bench to bedside. Clin Cancer Res. 2013;19(17):4801–4815. doi: 10.1158/1078-0432.CCR-13-1185. PubMed DOI PMC

Truxova I, Pokorna K, Kloudova K, Partlova S, Spisek R, Fucikova J. Day 3 Poly (I:C)-activated dendritic cells generated in CellGro for use in cancer immunotherapy trials are fully comparable to standard Day 5 DCs. Immunol Lett. 2014;160(1):39–49. doi: 10.1016/j.imlet.2014.03.010. PubMed DOI

Win SJ, McMillan DG, Errington-Mais F, Ward VK, Young SL, Baird MA, Melcher AA. Enhancing the immunogenicity of tumour lysate-loaded dendritic cell vaccines by conjugation to virus-like particles. Br J Cancer. 2012;106(1):92–98. doi: 10.1038/bjc.2011.538. PubMed DOI PMC

Schreiber H, Rowley JD, Rowley DA. Targeting mutations predictably. Blood. 2011;118(4):830–831. doi: 10.1182/blood-2011-06-357541. PubMed DOI

Lakshminarayanan V, Supekar NT, Wei J, McCurry DB, Dueck AC, Kosiorek HE, Trivedi PP, Bradley JM, Madsen CS, Pathangey LB, Hoelzinger DB, Wolfert MA, Boons GJ, Cohen PA, Gendler SJ. MUC1 vaccines, comprised of glycosylated or non-glycosylated peptides or tumor-derived MUC1, can circumvent immunoediting to control tumor growth in MUC1 transgenic mice. PLoS ONE. 2016;11(1):e0145920. doi: 10.1371/journal.pone.0145920. PubMed DOI PMC

Nicholaou T, Chen W, Davis ID, Jackson HM, Dimopoulos N, Barrow C, Browning J, Macgregor D, Williams D, Hopkins W, Maraskovsky E, Venhaus R, Pan L, Hoffman EW, Old LJ, Cebon J. Immunoediting and persistence of antigen-specific immunity in patients who have previously been vaccinated with NY-ESO-1 protein formulated in ISCOMATRIX. Cancer Immunol Immunother. 2011;60(11):1625–1637. doi: 10.1007/s00262-011-1041-3. PubMed DOI PMC

Berard F, Blanco P, Davoust J, Neidhart-Berard EM, Nouri-Shirazi M, Taquet N, Rimoldi D, Cerottini JC, Banchereau J, Palucka AK. Cross-priming of naive CD8 T cells against melanoma antigens using dendritic cells loaded with killed allogeneic melanoma cells. J Exp Med. 2000;192(11):1535–1544. doi: 10.1084/jem.192.11.1535. PubMed DOI PMC

Neidhardt-Berard EM, Berard F, Banchereau J, Palucka AK. Dendritic cells loaded with killed breast cancer cells induce differentiation of tumor-specific cytotoxic T lymphocytes. Breast Cancer Res. 2004;6(4):R322–R328. doi: 10.1186/bcr794. PubMed DOI PMC

Aguilar LK, Guzik BW, Aguilar-Cordova E. Cytotoxic immunotherapy strategies for cancer: mechanisms and clinical development. J Cell Biochem. 2011;112(8):1969–1977. doi: 10.1002/jcb.23126. PubMed DOI

Baxevanis CN, Voutsas IF, Tsitsilonis OE, Gritzapis AD, Sotiriadou R, Papamichail M. Tumor-specific CD4+ T lymphocytes from cancer patients are required for optimal induction of cytotoxic T cells against the autologous tumor. J Immunol. 2000;164(7):3902–3912. doi: 10.4049/jimmunol.164.7.3902. PubMed DOI

Quezada SA, Simpson TR, Peggs KS, Merghoub T, Vider J, Fan X, Blasberg R, Yagita H, Muranski P, Antony PA, Restifo NP, Allison JP. Tumor-reactive CD4+ T cells develop cytotoxic activity and eradicate large established melanoma after transfer into lymphopenic hosts. J Exp Med. 2010;207(3):637–650. doi: 10.1084/jem.20091918. PubMed DOI PMC

Aarntzen EH, De Vries IJ, Lesterhuis WJ, Schuurhuis D, Jacobs JF, Bol K, Schreibelt G, Mus R, De Wilt JH, Haanen JB, Schadendorf D, Croockewit A, Blokx WA, Van Rossum MM, Kwok WW, Adema GJ, Punt CJ, Figdor CG. Targeting CD4+ T-helper cells improves the induction of antitumor responses in dendritic cell-based vaccination. Cancer Res. 2013;73(1):19–29. doi: 10.1158/0008-5472.CAN-12-1127. PubMed DOI

Trombetta ES, Ebersold M, Garrett W, Pypaert M, Mellman I. Activation of lysosomal function during dendritic cell maturation. Science. 2003;299(5611):1400–1403. doi: 10.1126/science.1080106. PubMed DOI

Spadaro F, Lapenta C, Donati S, Abalsamo L, Barnaba V, Belardelli F, Santini SM, Ferrantini M. IFN-alpha enhances cross-presentation in human dendritic cells by modulating antigen survival, endocytic routing, and processing. Blood. 2012;119(6):1407–1417. doi: 10.1182/blood-2011-06-363564. PubMed DOI

Berk E, Xu S, Czerniecki BJ. Dendritic cells matured in the presence of TLR ligands overcome the immunosuppressive functions of regulatory T cells. Oncoimmunology. 2014;3:e27617. doi: 10.4161/onci.27617. PubMed DOI PMC

Brezar V, Ruffin N, Richert L, Surenaud M, Lacabaratz C, Palucka K, Thiebaut R, Banchereau J, Levy Y, Seddiki N. Decreased HIV-specific T-regulatory responses are associated with effective DC-vaccine induced immunity. PLoS Pathog. 2015;11(3):e1004752. doi: 10.1371/journal.ppat.1004752. PubMed DOI PMC

Santini SM, Lapenta C, Santodonato L, D’Agostino G, Belardelli F, Ferrantini M. IFN-alpha in the generation of dendritic cells for cancer immunotherapy. Handb Exp Pharmacol. 2009;188:295–317. doi: 10.1007/978-3-540-71029-5_14. PubMed DOI

Pantel A, Teixeira A, Haddad E, Wood EG, Steinman RM, Longhi MP. Direct type I IFN but not MDA5/TLR3 activation of dendritic cells is required for maturation and metabolic shift to glycolysis after poly IC stimulation. PLoS Biol. 2014;12(1):e1001759. doi: 10.1371/journal.pbio.1001759. PubMed DOI PMC