Recent studies have identified molecular events characteristic of immunogenic cell death (ICD), including surface exposure of calreticulin (CRT), the heat shock proteins HSP70 and HSP90, the release of high-mobility group box protein 1 (HMGB1) and the release of ATP from dying cells. We investigated the potential of high hydrostatic pressure (HHP) to induce ICD in human tumor cells. HHP induced the rapid expression of HSP70, HSP90 and CRT on the cell surface. HHP also induced the release of HMGB1 and ATP. The interaction of dendritic cells (DCs) with HHP-treated tumor cells led to a more rapid rate of DC phagocytosis, upregulation of CD83, CD86 and HLA-DR and the release of interleukin IL-6, IL-12p70 and TNF-α. DCs pulsed with tumor cells killed by HHP induced high numbers of tumor-specific T cells. DCs pulsed with HHP-treated tumor cells also induced the lowest number of regulatory T cells. In addition, we found that the key features of the endoplasmic reticulum stress-mediated apoptotic pathway, such as reactive oxygen species production, phosphorylation of the translation initiation factor eIF2α and activation of caspase-8, were activated by HHP treatment. Therefore, HHP acts as a reliable and potent inducer of ICD in human tumor cells.

• MeSH
• adenosintrifosfát sekrece   MeSH
• aktivace enzymů imunologie   MeSH
• antigeny CD86 biosyntéza   MeSH
• apoptóza imunologie   MeSH
• CD antigeny biosyntéza   MeSH
• dendritické buňky imunologie   MeSH
• eukaryotický iniciační faktor 2 metabolismus   MeSH
• fagocytóza imunologie   MeSH
• fosforylace MeSH
• HLA-DR antigeny biosyntéza   MeSH
• hydrostatický tlak MeSH
• imunoglobuliny biosyntéza   MeSH
• interleukin-12 sekrece   MeSH
• interleukin-6 sekrece   MeSH
• kalretikulin biosyntéza   imunologie   MeSH
• kaspasa 8 metabolismus   MeSH
• lidé MeSH
• membránové glykoproteiny biosyntéza   MeSH
• membránové proteiny biosyntéza   MeSH
• nádorové buněčné linie MeSH
• nádory imunologie   MeSH
• protein HMGB1 imunologie   sekrece   MeSH
• proteiny tepelného šoku HSP70 biosyntéza   imunologie   MeSH
• proteiny tepelného šoku HSP90 biosyntéza   imunologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulační T-lymfocyty imunologie   MeSH
• stres endoplazmatického retikula imunologie   MeSH
• TNF-alfa sekrece   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Z imunologického hlediska lze buněčnou smrt nádorových buněk rozlišit na imunogenní a neimunogenní v závislosti na podnětu, který ji vyvolává. Imunogenní buněčná smrt na rozdíl od fyziologické apoptózy vede ke stimulaci imunitního systému pomocí molekul, které jsou označovány jako DAMPs (danger associated molecular pattern). V prvních hodinách imunogenní buněčné apoptózy dochází k rychlé translokaci chaperonového proteinu kalretikulinu a proteinů teplotního stresu 70 a 90 (heat‑shock – HSP70 a HSP90) z endoplazmatického retikula na buněčný povrch. Následně dochází k aktivní sekreci molekuly adenosintrifosfátu a pasivnímu uvolnění nukleárního proteinu HMGB1 do extracelulárního prostoru. Společnou vlastností těchto molekul je aktivace buněk imunitního systému, zejména pak dendritických buněk, které se řadí mezi profesionální antigen prezentující buňky, které následně aktivují protinádorovou imunitní odpověď. Cílem tohoto souhrnného článku je popsat dosud známé induktory imunogenní buněčné smrti a jejich vliv na protinádorovou imunitní odpověď.

Cancer cell death can be immunogenic or nonimmunogenic depending on the initiating stimulus. The immunogenic characteristics of immunogenic cell death are mainly mediated by damage-associated molecular patterns represented by preapoptotic exposure of calreticulin and heat shock proteins (HSP70 and HSP90) from endoplasmic reticulum at the cell surface and active secretion of adenosintriphospate. Other damage-associated molecular patterns are produced in late stage apoptosis as high mobility group box 1 protein (HMGB1) into the extracellular milieu. Such signals operate on various receptors expressed by antigen presenting cells, mainly by population of dendritic cells, to stimulate the activation of antigen specific T-cell response. In this review, we describe the current known immunogenic cell death inducers and their potential to activate antitumor immune response. Key words: calreticulin – dendritic cell – immunogenic cell death – antitumor immune response This work was supported by grant IGA MH CZ – NT 12402-5 and MH CZ – DRO, University Hospital Motol, Prague, Czech Republic 00064203. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 29. 7. 2015 Accepted: 1. 10. 2015

• Klíčová slova
• protinádorová imunitní odpověď, induktory imunogenní smrti, coxackie viry CVB3, oxaliplatina,
• MeSH
• antracykliny farmakologie   imunologie   MeSH
• bortezomib MeSH
• buněčná imunita * imunologie   MeSH
• buněčná smrt * imunologie   MeSH
• cyklofosfamid farmakologie   imunologie   MeSH
• dendritické buňky MeSH
• fotochemoterapie MeSH
• hydrostatický tlak MeSH
• imunogenní buněčná smrt MeSH
• kalretikulin MeSH
• kyseliny boronové farmakologie   imunologie   MeSH
• lidé MeSH
• mitoxantron farmakologie   imunologie   MeSH
• nádory * imunologie   terapie   MeSH
• organoplatinové sloučeniny farmakologie   imunologie   MeSH
• pyraziny farmakologie   imunologie   MeSH
• radioterapie MeSH
• ultrafialové záření MeSH
• virové receptory MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

High hydrostatic pressure (HHP) induces immunogenic death of tumor cells which confer protective anti-tumor immunity in vivo. Moreover, DC pulsed with HHP-treated tumor cells induced therapeutic effect in mouse cancer model. In this study, we tested the immunogenicity, stability and T cell stimulatory activity of human monocyte-derived dendritic cell (DC)-based HHP lung cancer vaccine generated in GMP compliant serum free medium using HHP 250 MPa. DC pulsed with HHP-killed lung cancer cells and poly(I:C) enhanced DC maturation, chemotactic migration and production of pro-inflammatory cytokines after 24h. Moreover, DC-based HHP lung cancer vaccine showed functional plasticity after transfer into serum-containing media and stimulation with LPS or CD40L after additional 24h. LPS and CD40L stimulation further differentially enhanced the expression of costimulatory molecules and production of IL-12p70. DC-based HHP lung cancer vaccine decreased the number of CD4+CD25+Foxp3+ T regulatory cells and stimulated IFN-γ-producing tumor antigen-specific CD4+ and CD8+ T cells from non-small cell lung cancer (NSCLC) patients. Tumor antigen specific CD8+ and CD4+ T cell responses were detected in NSCLC patient's against a selected tumor antigens expressed by lung cancer cell lines used for the vaccine generation. We also showed for the first time that protein antigen from HHP-killed lung cancer cells is processed and presented by DC to CD8+ T cells. Our results represent important preclinical data for ongoing NSCLC Phase I/II clinical trial using DC-based active cellular immunotherapy (DCVAC/LuCa) in combination with chemotherapy and immune enhancers.

• MeSH
• CD4-pozitivní T-lymfocyty imunologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• dendritické buňky imunologie   MeSH
• hydrostatický tlak MeSH
• imunoterapie metody   MeSH
• interferon gama metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory plic terapie   MeSH
• nemalobuněčný karcinom plic terapie   MeSH
• protinádorové vakcíny imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

High hydrostatic pressure (HHP) has been shown to induce immunogenic cell death of cancer cells, facilitating their uptake by dendritic cells (DC) and subsequent presentation of tumor antigens. In the present study, we demonstrated immunogenicity of the HHP-treated tumor cells in mice. HHP was able to induce immunogenic cell death of both TC-1 and TRAMP-C2 tumor cells, representing murine models for human papilloma virus-associated tumors and prostate cancer, respectively. HHP-treated cells induced stronger immune responses in mice immunized with these tumor cells, documented by higher spleen cell cytotoxicity and increased IFNγ production as compared to irradiated tumor cells, accompanied by suppression of tumor growth in vivo in the case of TC-1 tumors, but not TRAMP-C2 tumors. Furthermore, HHP-treated cells were used for DC-based vaccine antigen pulsing. DC co-cultured with HHP-treated tumor cells and matured by a TLR 9 agonist exhibited higher cell surface expression of maturation markers and production of IL-12 and other cytokines, as compared to the DC pulsed with irradiated tumor cells. Immunization with DC cell-based vaccines pulsed with HHP-treated tumor cells induced high immune responses, detected by increased spleen cell cytotoxicity and elevated IFNγ production. The DC-based vaccine pulsed with HHP-treated tumor cells combined with docetaxel chemotherapy significantly inhibited growth of both TC-1 and TRAMP-C2 tumors. Our results indicate that DC-based vaccines pulsed with HHP-inactivated tumor cells can be a suitable tool for chemoimmunotherapy, particularly with regard to the findings that poorly immunogenic TRAMP-C2 tumors were susceptible to this treatment modality.

• MeSH
• antigeny nádorové metabolismus   MeSH
• antitumorózní látky aplikace a dávkování   MeSH
• cytotoxicita imunologická MeSH
• dendritické buňky cytologie   MeSH
• experimentální nádory farmakoterapie   terapie   MeSH
• hydrostatický tlak MeSH
• imunitní systém MeSH
• imunoterapie metody   MeSH
• infekce papilomavirem farmakoterapie   terapie   MeSH
• interferon gama metabolismus   MeSH
• interleukin-12 metabolismus   MeSH
• lidé MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory prostaty farmakoterapie   metabolismus   terapie   MeSH
• protinádorové vakcíny chemie   MeSH
• slezina imunologie   MeSH
• taxoidy aplikace a dávkování   MeSH
• toll-like receptor 9 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH