Blebs produced by actin–myosin contraction during apoptosis release damage-associated molecular pattern proteins before secondary necrosis occurs

Wickman, Grant; Julian, Linda; Mardilovich, Katerina; Schumacher, S.; Munro, June; Rath, Nicola; Zander, Serge A.L.; Mleczak, Andrzej; Sumpton, David; Morrice, Nicholas A.; Bienvenut, Willy V.; Olson, Michael F.

doi:10.1038/cdd.2013.69

Cited by 160 publications

(172 citation statements)

References 43 publications

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“…The modest increased uptake of blebs compared to ACR (14.8% versus 9.2%, respectively), and thereby the higher number of MART-1 [26][27][28][29][30][31][32][33][34][35] presenting skin DCs, is unlikely to be solely responsible for the selective activation of the TAA-specific CD8 C CTLs. It is known that heat shock proteins (HSPs), previously described to be present at high levels in apoptotic blebs, 35 greatly facilitate cross-presentation.…”

Section: 1332mentioning

confidence: 99%

“…It is known that heat shock proteins (HSPs), previously described to be present at high levels in apoptotic blebs, 35 greatly facilitate cross-presentation. 36 In our system, HSPs could, therefore, have been responsible for the observed enhancing effect on cross-presentation mediated by bleb-administered DCs.…”

Section: 1332mentioning

confidence: 99%

“…C and CD8 C T-cell proliferation (n D 7). (B) DCs that egressed from the skin biopsies after injecting either wild-type or MART-1 expressing apoptotic cell remnants (ACRs) or blebs, were co-cultured with a MART-1 [26][27][28][29][30][31][32][33][34][35] recognizing and HLA-A2 restricted cytotoxic T lymphocyte (CTL) line (> 95% pure) for 24 hours. Antigen-specific activation was determined by deducting the levels of interferon g (IFNg) produced following the injection of wild-type ACRs or blebs from the levels produced after injecting MART-1 expressing apoptotic fractions (n D 3).…”

Section: Intradermal Bleb or Acr Administration Does Not Interfere Wimentioning

confidence: 99%

“…We generated a CTL line recognizing MART-1 [26][27][28][29][30][31][32][33][34][35] in an HLA-A2-restricted fashion (>95% pure, as tested by dextramer staining), by priming CD8 C T cells from healthy donors with MART-1 26-35(27L) peptide-loaded JY cells. After stimulation, the expanded MART-1-specific T cells were sorted using a MART-1 [26][27][28][29][30][31][32][33][34][35] dextramer (Immudex).…”

Section: Antigen Cross-presentationmentioning

confidence: 99%

“…injected in skin explants from HLA-A2 positive donors in the presence of 4/GM, and skin DC were harvested after 48-72 hours. After a subsequent 24 hours of co-culture with a MART-1 [26][27][28][29][30][31][32][33][34][35] recognizing CTL line (HLA-A2 restricted; >95% pure), the supernatant was analyzed for the presence of IFNg, as a measure of cross-presentation of the MART-1 [26][27][28][29][30][31][32][33][34][35] epitope by egressed DC. The MART-1 [26][27][28][29][30][31][32][33][34][35] specific IFNg production was assessed by deducting the IFNg production following a co-culture with DC from wild-type ACR or blebs injected skin, from that secreted after co-culturing with DC from MART-1-transduced ACR or blebs (D IFNg, Fig.…”

Section: Intradermal Bleb or Acr Administration Does Not Interfere Wimentioning

confidence: 99%

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In situloading of skin dendritic cells with apoptotic bleb-derived antigens for the induction of tumor-directed immunity

et al. 2014

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The generation and loading of dendritic cells (DC) ex-vivo for tumor vaccination purposes is laborious and costly. Direct intradermal (i.d.) administration of tumor-associated antigens could be an attractive alternative approach, provided that efficient uptake and cross-presentation by appropriately activated skin DCs can be achieved. Here, we compare the efficiency of i.d. delivery of relatively small apoptotic blebs (diameter »0.1-1 mm) derived from MART-1 transduced acute myeloid leukemia (AML) HL60 cells, to that of larger apoptotic cell remnants (ACR; 2-10 mm) in a physiologically highly relevant human skin explant model. Injection of either fluorescently-labelled ACRs or blebs alone did not affect the number or distribution of migrated DC subsets from skin biopsies after 48 hours, but resulted in a general up-regulation of the co-stimulatory molecules CD83 and CD86 on skin DCs that had ingested apoptotic material. We have previously shown that i.d. administration of GM-CSF and IL-4 resulted in preferential migration of a mature and highly T cell-stimulatory CD11 hi CD1a C CD14 ¡ dermal DC subset. Here, we found that co-injection of GM-CSF and IL-4 together with either ACRs or blebs resulted in uptake efficiencies within this dermal DC subset of 7.6% ( §6.1%) and 19.1% ( §15.9%), respectively, thus revealing a significantly higher uptake frequency of blebs (P < 0.02). Intradermal delivery of tumor-derived blebs did not affect the T-cell priming and T H -skewing abilities of migratory skin DC. Nevertheless, in contrast to i.d. administration of ACR, the injection of blebs lead to effective cross-presentation of MART-1 to specific CD8 C effector T cells. We conclude that apoptotic bleb-based vaccines delivered through the skin may offer an attractive, and broadly applicable, cancer immunotherapy.

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Section: 1332mentioning

confidence: 99%

Section: 1332mentioning

confidence: 99%

Section: Intradermal Bleb or Acr Administration Does Not Interfere Wimentioning

confidence: 99%

Section: Antigen Cross-presentationmentioning

confidence: 99%

Section: Intradermal Bleb or Acr Administration Does Not Interfere Wimentioning

confidence: 99%

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In situloading of skin dendritic cells with apoptotic bleb-derived antigens for the induction of tumor-directed immunity

et al. 2014

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Dismantling the Apoptotic Cell by Caspases

Deming

Kurokawa

2017

Encyclopedia of Life Sciences

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Apoptosis is a programme of cell death that results in dramatic morphological and biochemical changes in the dying cell due to the systematic dismantling of cellular architecture and functional pathways. The proteins that execute the apoptotic programme are a group of proteases termed caspases (cysteine‐dependent aspartate‐specific protease). Caspases proteolytically cleave a host of cellular substrates at aspartate residues, which may render them either functionally inactive or confer novel activities that help to promote cellular demise. Substrates targeted by caspases during the apoptotic programme include proteins involved in maintaining various aspects of cytoskeletal and organelle architecture as well as proteins that function in signalling networks critical for cell function. Following the execution phase of apoptosis, the cellular corpse is packaged in an orderly fashion into membrane‐bound apoptotic bodies that are sensed by phagocytes, which neatly engulf the dead cell without eliciting an immune response. Key Concepts Apoptotic caspases are cysteine proteases that become activated in response to diverse extracellular and intracellular stimuli and subsequently carry out the cell death programme by systematically cleaving intracellular proteins. A hierarchy of caspase activation exists whereby initiator caspases become activated to cleave and activate effector caspases. The cellular morphological changes associated with apoptosis encompass three stages: release, membrane blebbing and condensation. Caspase‐mediated cleavage of target proteins may produce stable functional effector fragments or unstable fragments that are quickly degraded. Caspases target multiple aspects of the cellular architecture to induce collapse of organelles and the cytoskeleton. Signalling networks that regulate cellular processes critical for cell survival are inactivated by caspases. As the dying cell is dismantled during the apoptotic process, it orchestrates its own disposal by displaying ‘eat me’ signals on its cell surface and releasing ‘find me’ signals to recruit phagocytic cells.

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Transient Support from Fibroblasts is Sufficient to Drive Functional Vascularization in Engineered Tissues

Song

Lammers

Sundaram

et al. 2020

Adv Funct Materials

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Formation of capillary blood vasculature is a critical requirement for native as well as engineered organs and can be induced in vitro by coculturing endothelial cells with fibroblasts. However, whether these fibroblasts are required only in the initial morphogenesis of endothelial cells or needed throughout is unknown, and the ability to remove these stromal cells after assembly can be useful for clinical translation. In this study, a technique termed CAMEO (Controlled Apoptosis in Multicellular Tissues for Engineered Organogenesis) is introduced, whereby fibroblasts are selectively ablated on demand, and it is utilized to probe the dispensability of fibroblasts in vascular morphogenesis. The presence of fibroblasts is shown to be necessary only during the first few days of endothelial cell morphogenesis, after which they can be ablated without significantly affecting the structural and functional features of the developed vasculature. Furthermore, the use of CAMEO to vascularize a construct containing primary human hepatocytes that improved tissue function is demonstrated. In conclusion, this study suggests that transient, initial support from fibroblasts is sufficient to drive vascular morphogenesis in engineered tissues, and this strategy of engineering-via-elimination may provide a new general approach for achieving desired functions and cell compositions in engineered organs.

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Blebs produced by actin–myosin contraction during apoptosis release damage-associated molecular pattern proteins before secondary necrosis occurs

Cited by 160 publications

References 43 publications

In situloading of skin dendritic cells with apoptotic bleb-derived antigens for the induction of tumor-directed immunity

In situloading of skin dendritic cells with apoptotic bleb-derived antigens for the induction of tumor-directed immunity

Dismantling the Apoptotic Cell by Caspases

Transient Support from Fibroblasts is Sufficient to Drive Functional Vascularization in Engineered Tissues

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