In vitro Evaluation of Programmed Cell Death in the Immune System of Pacific Oyster Crassostrea gigas by the Effect of Marine Toxins

Programmed cell death (PCD) is an essential process for the immune system's development and homeostasis, enabling the remotion of infected or unnecessary cells. There are several PCD's types, depending on the molecular mechanisms, such as non-inflammatory or pro-inflammatory. Hemocytes are the main component of cellular immunity in bivalve mollusks. Numerous infectious microorganisms produce toxins that impair hemocytes functions, but there is little knowledge on the role of PCD in these cells. This study aims to evaluate in vitro whether marine toxins induce a particular type of PCD in hemocytes of the bivalve mollusk Crassostrea gigas during 4 h at 25°C. Hemocytes were incubated with two types of marine toxins: non-proteinaceous toxins from microalgae (saxitoxin, STX; gonyautoxins 2 and 3, GTX2/3; okadaic acid/dynophysistoxin-1, OA/DTX-1; brevetoxins 2 and 3, PbTx-2,-3; brevetoxin 2, PbTx-2), and proteinaceous extracts from bacteria (Vibrio parahaemolyticus, Vp; V. campbellii, Vc). Also, we used the apoptosis inducers, staurosporine (STP), and camptothecin (CPT). STP, CPT, STX, and GTX 2/3, provoked high hemocyte mortality characterized by apoptosis hallmarks such as phosphatidylserine translocation into the outer leaflet of the cell membrane, exacerbated chromatin condensation, DNA oligonucleosomal fragments, and variation in gene expression levels of apoptotic caspases 2, 3, 7, and 8. The mixture of PbTx-2,-3 also showed many apoptosis features; however, they did not show apoptotic DNA oligonucleosomal fragments. Likewise, PbTx-2, OA/DTX-1, and proteinaceous extracts from bacteria Vp, and Vc, induced a minor degree of cell death with high gene expression of the pro-inflammatory initiator caspase-1, which could indicate a process of pyroptosis-like PCD. Hemocytes could carry out both PCD types simultaneously. Therefore, marine toxins trigger PCD's signaling pathways in C. gigas hemocytes, depending on the toxin's nature, which appears to be highly conserved both structurally and functionally.

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Programmed Cell Death-1: Programmed Cell Death-Ligand 1 Interaction Protects Human Cardiomyocytes Against T-Cell Mediated Inflammation and Apoptosis Response In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms21072399 ◽

2020 ◽

Vol 21 (7) ◽

pp. 2399

Author(s):

Woan Ting Tay ◽

Yi-Hsien Fang ◽

Suet Theng Beh ◽

Yen-Wen Liu ◽

Ling-Wei Hsu ◽

...

Keyword(s):

Gene Expression ◽

Cell Death ◽

T Cell ◽

T Lymphocytes ◽

Programmed Cell Death ◽

T Cell Response ◽

Tumor Bearing ◽

Cd8 T Lymphocytes ◽

Programmed Cell Death 1

Aim: Immunological checkpoint therapy is considered a powerful method for cancer therapy and acts by re-activating autologous T cells to kill the cancer cell. Myocarditis cases have been reported in cancer patients after immunological therapy; for example, nivolumab treatment is a monoclonal antibody that blocks programmed cell death-1/programmed cell death ligand-1 ligand interaction. This project provided insight into the inflammatory response as a benchmark to investigate the potential cardiotoxic effect of T cell response to the programmed cell death-1 (PD-1)/programmed cell death ligand-1 (PD-L1) axis in regulating cardiomyocyte injury in vitro. Methods and Results: We investigated cardiomyopathy resulted from the PD-1/PD-L1 axis blockade using the anti-PD-1 antibody in Rockefeller University embryonic stem cells-derived cardiomyocytes (RUES2-CMs) and a melanoma tumor-bearing murine model. We found that nivolumab alone did not induce inflammatory-related proteins, including PD-L1 expression, and did not induce apoptosis, which was contrary to doxorubicin, a cardiotoxic chemotherapy drug. However, nivolumab was able to exacerbate the immune response by increasing cytokine and inflammatory gene expression in RUES2-CMs when co-cultured with CD4+ T lymphocytes and induced apoptosis. This effect was not observed when RUES2-CMs were co-cultured with CD8+ T lymphocytes. The in vivo model showed that the heart function of tumor-bearing mice was decreased after treatment with anti-PD-1 antibody and demonstrated a dilated left ventricle histological examination. The dilated left ventricle was associated with an infiltration of CD4+ and CD8+ T lymphocytes into the myocardium. PD-L1 and inflammatory-associated gene expression were significantly increased in anti-PD-1-treated tumor-bearing mice. Cleaved caspase-3 and mouse plasma cardiac troponin I expressions were increased significantly. Conclusion: PD-L1 expression on cardiomyocytes suppressed T-cell function. Blockade of PD-1 by nivolumab enhanced cardiomyocyte inflammation and apoptosis through the enhancement of T-cell response towards cardiomyocytes.

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Programmed cell death in Dictyostelium

Journal of Cell Science ◽

10.1242/jcs.107.10.2691 ◽

1994 ◽

Vol 107 (10) ◽

pp. 2691-2704 ◽

Cited By ~ 10

Author(s):

S. Cornillon ◽

C. Foa ◽

J. Davoust ◽

N. Buonavista ◽

J.D. Gross ◽

...

Keyword(s):

Cell Death ◽

Programmed Cell Death ◽

Chromatin Condensation ◽

In Vitro System ◽

The Core ◽

Mechanical Constraints ◽

Propidium Iodide Staining ◽

Temporal Succession ◽

Phenotypic Variations

Programmed cell death (PCD) of Dictyostelium discoideum cells was triggered precisely and studied quantitatively in an in vitro system involving differentiation without morphogenesis. In temporal succession after the triggering of differentiation, PCD included first an irreversible step leading to the inability to regrow at 8 hours. At 12 hours, massive vacuolisation was best evidenced by confocal microscopy, and prominent cytoplasmic condensation and focal chromatin condensation could be observed by electron microscopy. Membrane permeabilization occurred only very late (at 40–60 hours) as judged by propidium iodide staining. No early DNA fragmentation could be detected by standard or pulsed field gel electrophoresis. These traits exhibit some similarity to those of previously described non-apoptotic and apoptotic PCD, suggesting the hypothesis of a single core molecular mechanism of PCD emerging in evolution before the postulated multiple emergences of multicellularity. A single core mechanism would underly phenotypic variations of PCD resulting in various cells from differences in enzymatic equipment and mechanical constraints. A prediction is that some of the molecules involved in the core PCD mechanism of even phylogenetically very distant organisms, e.g. Dictyostelium and vertebrates, should be related.

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Altered Expression of Immune Checkpoint Molecules Including Programmed Cell Death-1 (PD-1) and Its Ligands PD-L1/PD-L2 in Waldenstrom's Macroglobulinemia

Blood ◽

10.1182/blood.v128.22.1772.1772 ◽

2016 ◽

Vol 128 (22) ◽

pp. 1772-1772 ◽

Cited By ~ 1

Author(s):

Shahrzad Jalali ◽

Tammy Price-Troska ◽

Anne J. Novak ◽

Stephen M Ansell

Keyword(s):

Bone Marrow ◽

T Cells ◽

Cell Death ◽

Programmed Cell Death ◽

B Cell ◽

Immune Checkpoint ◽

Molecular Mechanisms ◽

Immune Checkpoint Molecules ◽

Programmed Cell Death 1

Abstract Waldenstrom's Macroglobulinemia (WM) is an indolent B-cell malignancy that is characterized by bone marrow infiltration of lymphoplasmacytic cells and excessive synthesis of IgM. Despite improved treatment strategies and increased understanding of the underlying molecular mechanisms contributing to malignant disease progression, WM remains incurable. While whole exome sequencing analysis has revealed the genomic events associated with malignant B cell transformation in WM, the bone marrow (BM) microenvironment is also emerging as a major player in the pathogenesis of WM. Previously, our lab has reported that an array of cytokines, including IL-21 and IL-6 are increased in the WM BM and such increases are associated with high proliferation rates and IgM secretion by WM cells. In this study, we aimed to identify how immune checkpoint molecules in the BM microenvironment, specifically programmed cell death-1 (PD-1) and its ligands PD-L1 and PD-L2, contribute to malignant cell expansion in WM. Using Real-Time PCR analysis, we show that the expression of PD-1, PD-L1 and PD-L2 are all significantly elevated in the cells of WM BM as compared to their normal BM counterparts. Interestingly, PD-L1 expression is also upregulated in the malignant WM cells. These findings were further confirmed by flow cytometry analysis. Subsequent in vitro analysis showed that PD-L1 and PD-L2 are cleaved from the WM cells and these soluble ligands (sPD-L1 and sPD-L2) are able to bind to the cells in the BM microenvironment, including T-cells. This results in reduced activation and proliferation of T-cells, and is accompanied by a reduction in cyclin A and p-ERK levels. In validation of the in vitro data, we also found sPD-L1 and sPD-L2 in the bone marrow plasma obtained from WM patients. While sPD-L2 was found to be present in both WM and normal BM plasma (though in a reduced level in WM), sPD-L1 was exclusive to the WM samples, implying that the function of these molecules are differentially regulated in WM. Furthermore, overexpression of PD-L1 in a WM cell line, MWCL.1, resulted in an increase in IgM secretion. Interestingly, treatment with IL-21 increased the expression of PD-L1 in WM cells, indicating that elevated IL-21 in WM BM could contribute to increased expression of PD-L1 by malignant WM cells. In summary, our data support a role for PD-1 ligands as immune regulatory molecules in WM. Further studies are ongoing to explore the molecular mechanisms by which PD-1/PD-L1/PD-L2 signaling affects the pathophysiology of WM cells. Disclosures No relevant conflicts of interest to declare.

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In vitro activity of 1H-phenalen-1-one derivatives against Leishmania spp. and evidence of programmed cell death

Parasites & Vectors ◽

10.1186/s13071-019-3854-4 ◽

2019 ◽

Vol 12 (1) ◽

Cited By ~ 1

Author(s):

Atteneri López-Arencibia ◽

María Reyes-Batlle ◽

Mónica B. Freijo ◽

Ines Sifaoui ◽

Carlos J. Bethencourt-Estrella ◽

...

Keyword(s):

Cell Death ◽

Membrane Potential ◽

Programmed Cell Death ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Chromatin Condensation ◽

Death Process ◽

Leishmania Spp ◽

Characteristic Features

Abstract Background The in vitro activity against Leishmania spp. of a novel group of compounds, phenalenone derivatives, is described in this study. Previous studies have shown that some phenalenones present leishmanicidal activity, and induce a decrease in the mitochondrial membrane potential in L. amazonensis parasites, so in order to elucidate the evidence of programmed cell death occurring inside the promastigote stage, different assays were performed in two different species of Leishmania. Methods We focused on the determination of the programmed cell death evidence by detecting the characteristic features of the apoptosis-like process, such as phosphatidylserine exposure, mitochondrial membrane potential, and chromatin condensation among others. Results The results showed that four molecules activated the apoptosis-like process in the parasite. All the signals observed were indicative of the death process that the parasites were undergoing. Conclusions The present results highlight the potential use of phenalenone derivatives against Leishmania species and further studies should be undertaken to establish them as novel leishmanicidal therapeutic agents.

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Monitoring programmed cell death triggered by mild heat shock in soybean-cultured cells

Functional Plant Biology ◽

10.1071/fp06015 ◽

2006 ◽

Vol 33 (7) ◽

pp. 617 ◽

Cited By ~ 23

Author(s):

Anna Zuppini ◽

Valentina Bugno ◽

Barbara Baldan

Keyword(s):

Cell Death ◽

Heat Shock ◽

Programmed Cell Death ◽

Molecular Mechanisms ◽

Cultured Cells ◽

Chromatin Condensation ◽

H2o2 Production ◽

Cytochrome C Release ◽

Mild Heat ◽

Glycine Max L

Programmed cell death (PCD) is a common form of cellular demise during plant response to environmental stresses. The pathway of PCD has been partially clarified in plants although the underlying molecular mechanisms are still poorly defined. We have investigated the signalling cascade induced by a mild heat treatment causing PCD in soybean cells (Glycine max L.). The data show that heat shock led to the onset of PCD in soybean cells involving H2O2 production and mitochondrial damage. Cytochrome c release accompanies the presence of caspase 9-like and caspase 3-like protease activities. Concomitantly, cells were severely damaged with a progressive cell shrinkage, chloroplast alteration and detachment of the plasma membrane from the cell wall. Chromatin condensation and DNA damage were observed. It is proposed that a mild heat stress induces PCD in soybean cells through a caspase-like-dependent pathway.

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Human clusterin gene expression is confined to surviving cells during in vitro programmed cell death.

Journal of Clinical Investigation ◽

10.1172/jci117043 ◽

1994 ◽

Vol 93 (2) ◽

pp. 877-884 ◽

Cited By ~ 113

Author(s):

L E French ◽

A Wohlwend ◽

A P Sappino ◽

J Tschopp ◽

J A Schifferli

Keyword(s):

Gene Expression ◽

Cell Death ◽

Programmed Cell Death

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High-resolution analysis of differential gene expression during skeletal muscle atrophy and programmed cell death

Physiological Genomics ◽

10.1152/physiolgenomics.00047.2020 ◽

2020 ◽

Vol 52 (10) ◽

pp. 492-511

Author(s):

Junko Tsuji ◽

Travis Thomson ◽

Elizabeth Chan ◽

Christine K. Brown ◽

Julia Oppenheimer ◽

...

Keyword(s):

Gene Expression ◽

Cell Death ◽

Programmed Cell Death ◽

Molecular Mechanisms ◽

Cell Types ◽

Skeletal Muscle Atrophy ◽

Wide Range ◽

Cord Injury ◽

Associated Proteins ◽

Loss Of Mass

Skeletal muscles can undergo atrophy and/or programmed cell death (PCD) during development or in response to a wide range of insults, including immobility, cachexia, and spinal cord injury. However, the protracted nature of atrophy and the presence of multiple cell types within the tissue complicate molecular analyses. One model that does not suffer from these limitations is the intersegmental muscle (ISM) of the tobacco hawkmoth Manduca sexta. Three days before the adult eclosion (emergence) at the end of metamorphosis, the ISMs initiate a nonpathological program of atrophy that results in a 40% loss of mass. The ISMs then generate the eclosion behavior and initiate a nonapoptotic PCD during the next 30 h. We have performed a comprehensive transcriptomics analysis of all mRNAs and microRNAs throughout ISM development to better understand the molecular mechanisms that mediate atrophy and death. Atrophy involves enhanced protein catabolism and reduced expression of the genes involved in respiration, adhesion, and the contractile apparatus. In contrast, PCD involves the induction of numerous proteases, DNA methylases, membrane transporters, ribosomes, and anaerobic metabolism. These changes in gene expression are largely repressed when insects are injected with the insect steroid hormone 20-hydroxyecdysone, which delays death. The expression of the death-associated proteins may be greatly enhanced by reductions in specific microRNAs that function to repress translation. This study not only provides fundamental new insights into basic developmental processes, it may also represent a powerful resource for identifying potential diagnostic markers and molecular targets for therapeutic intervention.

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Molecular signature of anastasis for reversal of apoptosis

F1000Research ◽

10.12688/f1000research.10568.1 ◽

2017 ◽

Vol 6 ◽

pp. 43 ◽

Cited By ~ 12

Author(s):

Ho Man Tang ◽

C. Conover Talbot Jr ◽

Ming Chiu Fung ◽

Ho Lam Tang

Keyword(s):

Gene Expression ◽

Cell Death ◽

Time Course ◽

Molecular Mechanisms ◽

Expression Profiles ◽

Molecular Signature ◽

Cell Recovery ◽

Therapeutic Implications

Apoptosis is a type of programmed cell death that is essential for normal organismal development and homeostasis of multicellular organisms by eliminating unwanted, injured, or dangerous cells. This cell suicide process is generally assumed to be irreversible. However, accumulating studies suggest that dying cells can recover from the brink of cell death. We recently discovered an unexpected reversibility of the execution-stage of apoptosis in vitro and in vivo, and proposed the term anastasis (Greek for “rising to life”) to describe this cell recovery phenomenon. Promoting anastasis could in principle preserve injured cells that are difficult to replace, such as cardiomyocytes and neurons. Conversely, arresting anastasis in dying cancer cells after cancer therapies could improve treatment efficacy. To develop new therapies that promote or inhibit anastasis, it is essential to identify the key regulators and mediators of anastasis – the therapeutic targets. Therefore, we performed time-course microarray analysis to explore the molecular mechanisms of anastasis during reversal of ethanol-induced apoptosis in mouse primary liver cells. We found striking changes in transcription of genes involved in multiple pathways, including early activation of pro-survival genes, cell cycle arrest, stress-inducible responses, and at delayed times, cell migration and angiogenesis. Here, we present the time-course whole-genome gene expression dataset revealing gene expression profiles during the reversal of apoptosis. This dataset provides important insights into the physiological, pathological, and therapeutic implications of anastasis.

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Acrylonitrile Derivatives against Trypanosoma cruzi: In Vitro Activity and Programmed Cell Death Study

Pharmaceuticals ◽

10.3390/ph14060552 ◽

2021 ◽

Vol 14 (6) ◽

pp. 552

Author(s):

Carlos J. Bethencourt-Estrella ◽

Samuel Delgado-Hernández ◽

Atteneri López-Arencibia ◽

Desirée San Nicolás-Hernández ◽

Ines Sifaoui ◽

...

Keyword(s):

Cell Death ◽

Programmed Cell Death ◽

Trypanosoma Cruzi ◽

Chagas Disease ◽

Therapeutic Potential ◽

Chromatin Condensation ◽

Protozoan Parasite ◽

The United States ◽

Death Process

The neglected infection known as Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, results in more than 7000 deaths per year, with an increasing number of cases in non-endemic areas such as Europe or the United States. Moreover, with the current available therapy, only two compounds which are active against the acute phase of the disease are readily available. In addition, these therapeutic agents display multiple undesired side effects such as high toxicity, they are expensive, the treatment is lengthy and the resistant strain has emerged. Therefore, there is a need to find new compounds against Chagas disease which should be active against the parasite but also cause low toxicity to the patients. In the present work, the activity of novel acrylonitriles against Trypanosoma cruzi was evaluated as well as the analysis of the physiological events induced in the treated parasites related to the cell death process. Hence, the characteristic features of an apoptosis-like process such as chromatin condensation and mitochondrial membrane potential, among others, were studied. From the 32 compounds tested against the epimastigote stage of T. cruzi, 11 were selected based on their selectivity index to determine if these compounds were able to induce programmed cell death (PCD) in the treated parasites. Furthermore, acrylonitriles Q5, Q7, Q19, Q27 and Q29 were shown to trigger physiological events related in the PCD. Therefore, this study highlights the therapeutic potential of acrylonitriles as novel trypanocidal agents.

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