Combination of ionising radiation with hyperthermia increases the immunogenic potential of B16-F10 melanoma cells<i>in vitro</i>and<i>in vivo</i>

Werthmöller, Nina; Frey, Benjamin; Rückert, Michael; Lotter, Michael; Fietkau, Rainer; Gaipl, Udo S.

doi:10.3109/02656736.2015.1106011

Cited by 59 publications

(58 citation statements)

References 67 publications

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“…An increase was detected in the fraction of splenic DCs expressing TLR4 in the directly irradiated cells especially after irradiation with 2 Gy. This is in line with published data demonstrating that irradiation leads to increased release of danger signals, such as HMGB1, which interact with DCs via their TLR4 receptor (66–68). However, in bystander animals, the fraction of TLR4-expressing DCs decreased to half of the control level and changes were not influenced by radiation dose.…”

Section: Discussionsupporting

confidence: 92%

Extracellular Vesicles Mediate Radiation-Induced Systemic Bystander Signals in the Bone Marrow and Spleen

et al. 2017

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Radiation-induced bystander effects refer to the induction of biological changes in cells not directly hit by radiation implying that the number of cells affected by radiation is larger than the actual number of irradiated cells. Recent in vitro studies suggest the role of extracellular vesicles (EVs) in mediating radiation-induced bystander signals, but in vivo investigations are still lacking. Here, we report an in vivo study investigating the role of EVs in mediating radiation effects. C57BL/6 mice were total-body irradiated with X-rays (0.1, 0.25, 2 Gy), and 24 h later, EVs were isolated from the bone marrow (BM) and were intravenously injected into unirradiated (so-called bystander) animals. EV-induced systemic effects were compared to radiation effects in the directly irradiated animals. Similar to direct radiation, EVs from irradiated mice induced complex DNA damage in EV-recipient animals, manifested in an increased level of chromosomal aberrations and the activation of the DNA damage response. However, while DNA damage after direct irradiation increased with the dose, EV-induced effects peaked at lower doses. A significantly reduced hematopoietic stem cell pool in the BM as well as CD4+ and CD8+ lymphocyte pool in the spleen was detected in mice injected with EVs isolated from animals irradiated with 2 Gy. These EV-induced alterations were comparable to changes present in the directly irradiated mice. The pool of TLR4-expressing dendritic cells was different in the directly irradiated mice, where it increased after 2 Gy and in the EV-recipient animals, where it strongly decreased in a dose-independent manner. A panel of eight differentially expressed microRNAs (miRNA) was identified in the EVs originating from both low- and high-dose-irradiated mice, with a predicted involvement in pathways related to DNA damage repair, hematopoietic, and immune system regulation, suggesting a direct involvement of these pathways in mediating radiation-induced systemic effects. In conclusion, we proved the role of EVs in transmitting certain radiation effects, identified miRNAs carried by EVs potentially responsible for these effects, and showed that the pattern of changes was often different in the directly irradiated and EV-recipient bystander mice, suggesting different mechanisms.

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Section: Discussionsupporting

confidence: 92%

Extracellular Vesicles Mediate Radiation-Induced Systemic Bystander Signals in the Bone Marrow and Spleen

et al. 2017

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“…Tumors were irradiated with a dose of 5 Gy each day using a linear accelerator unit with 6 MV and a focus-skin distance of 1,000 mm. In order to protect healthy tissue, the gantry of the linear accelerator was rotated to 340° as previously described by our group (23). …”

Section: Methodsmentioning

confidence: 99%

Hypofractionated Irradiation Has Immune Stimulatory Potential and Induces a Timely Restricted Infiltration of Immune Cells in Colon Cancer Tumors

et al. 2017

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In addition to locally controlling the tumor, hypofractionated radiotherapy (RT) particularly aims to activate immune cells in the RT-modified microenvironment. Therefore, we examined whether hypofractionated RT can activate dendritic cells (DCs), induce immune cell infiltration in tumors, and how the chronology of immune cell migration into tumors occurs to gain knowledge for future definition of radiation breaks and inclusion of immunotherapy. Colorectal cancer treatments offer only limited survival benefit, and immunobiological principles for additional therapies need to be explored with preclinical models. The impact of hypofractionated RT on CT26 colon cancer tumor cell death, migration of DCs toward supernatants (SN) of tumor cells, and activation of DCs by SN were analyzed. The subcutaneous tumor of a BALB/c-CT26 mouse model was locally irradiated with 2 × 5 Gy, the tumor volume was monitored, and the infiltration of immune cells in the tumor was determined by flow cytometry daily. Hypofractionated RT induced a mixture of apoptotic and necrotic CT26 cells, which is known to be in particular immunogenic. DCs that migrated toward SN of CT26 cells particularly upregulated the activation markers CD80 and CD86 when in contact with SN of irradiated tumor cells. After hypofractionated RT, the tumor outgrowth was significantly retarded and in the irradiated tumors an increased infiltration of macrophages (CD11bhigh/F4-80+) and DCs (MHC-II+), but only between day 5 and 10 after the first irradiation, takes place. While CD4+ T cells migrated into non-irradiated and irradiated tumors, CD8+ T cells were only found in tumors that had been irradiated and they were highly increased at day 8 after the first irradiation. Myeloid-derived suppressor cells and regulatory T cells show regular turnover in irradiated and non-irradiated tumors. Tumor cell-specific anti-IgM antibodies were enhanced in the serum of animals with irradiated tumors. We conclude that hypofractionated RT suffices to activate DCs and to induce infiltration of innate and adaptive immune cells into solid colorectal tumors. However, the presence of immune cells in the tumor which are beneficial for antitumor immune responses is timely restricted. These findings should be considered when innovative multimodal tumor treatment protocols of distinct RT with immune therapies are designed and clinically implemented.

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“…Lethal RT doses (in particular when delivered according to a fractionated schedule) promote a variant of RCD that is highly immunogenic owing to the abundant and timely release of endogenous adjuvant-like molecules collectively known as damage-associated molecular patterns (DAMPs). 13,37,38 These DAMPs include (but may not be limited to): (i) CALR and other ER chaperones that become exposed on the plasma membrane of dying cells, [39][40][41] where they act as a potent phagocytic signals for dendritic cells (DCs) upon binding to LDL receptor-related protein 1 (LRP1, best known as CD91); 42 (ii) extracellular ATP, which is secreted by cancer cells succumbing to irradiation in an autophagy-dependent manner 40,43,44 and promotes tumor infiltration by myeloid cells-upon binding to purinergic receptor P2Y2 (P2RY2)-as well as their activation-upon binding to purinergic receptor P2X 7 (P2RX7); 45,46 (iii) extracellular high mobility group box 1 (HMGB1), 40,43 which mediates immunostimulatory effects in an advanced glycosylation end-product specific receptor (AGER)-and Toll-like receptor 4 (TLR4)-dependent manner; 47,48 and (iv) type I interferon (IFN), which operates in a dual mode to favor the release of chemotactic cytokines like C-X-C motif chemokine ligand 10 (CXCL10) from cancer cells (via an autocrine/paracrine mechanism) 49 and to promote the maturation/activation of tumor-infiltrating myeloid cells. 36 F I G U R E 1 On-target immunological effects of radiation therapy.…”

Section: Immunostimulationmentioning

confidence: 99%

Immune recognition of irradiated cancer cells

Wennerberg

Vanpouille-Box

Bornstein

et al. 2017

Immunological Reviews

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Summary Ionizing irradiation has been extensively employed for the clinical management of solid tumors, with therapeutic or palliative intents, for decades. Until recently, radiation therapy (RT) was believed to mediate antineoplastic activity mostly (if not only) as a consequence of cancer cell-intrinsic effects. Indeed, the macromolecular damage imposed to malignant cells by RT initiates one or multiple signal transduction cascades that drive a permanent proliferative arrest (cellular senescence) or regulated cell death. Both these phenomena show a rather linear dose-response correlation. However, RT also mediates consistent immunological activity, not only as an “on-target effect” originating within irradiated cancer cells, but also as an “off-target effect” depending on the interaction between RT and stromal, endothelial and immune components of the tumor microenvironment. Interestingly, the immunological activity of RT does not exhibit linear dose-response correlation. Here, we discuss the mechanisms whereby RT alters the capacity of the immune system to recognize and eliminate irradiated cancer cells, either as an “on-target” or as on “off-target” effect. In particular, we discuss the antagonism between the immunostimulatory and immunosuppressive effects of RT as we delineate combinatorial strategies to boost the former at the expenses of the latter.

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Combination of ionising radiation with hyperthermia increases the immunogenic potential of B16-F10 melanoma cellsin vitroandin vivo

Cited by 59 publications

References 67 publications

Extracellular Vesicles Mediate Radiation-Induced Systemic Bystander Signals in the Bone Marrow and Spleen

Extracellular Vesicles Mediate Radiation-Induced Systemic Bystander Signals in the Bone Marrow and Spleen

Hypofractionated Irradiation Has Immune Stimulatory Potential and Induces a Timely Restricted Infiltration of Immune Cells in Colon Cancer Tumors

Immune recognition of irradiated cancer cells

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