scholarly journals Adenovirus-Mediated FasL Minigene Transfer Endows Transduced Cells with Killer Potential

2020 ◽  
Vol 21 (17) ◽  
pp. 6011 ◽  
Author(s):  
Madalina Dumitrescu ◽  
Violeta Georgeta Trusca ◽  
Lorand Savu ◽  
Ioana Georgeta Stancu ◽  
Attila Cristian Ratiu ◽  
...  
Keyword(s):  
Fas Ligand ◽  
Type I Diabetes ◽  
Adenovirus Vector ◽  
Type I ◽  
Fasl Expression ◽  
Tnf Superfamily ◽  
Size Limitation ◽  
Fasl Gene ◽  
Induced Apoptosis ◽  
Necrosis Factor

Fas ligand (First apoptosis signal ligand, FasL, also known as CD95L) is the common executioner of apoptosis within the tumor necrosis factor (TNF) superfamily. We aimed to induce functional FasL expression in transduced cells using an adenovirus vector, which has the advantage of strong and transient induction of the gene included in the adenoviral genome. Here, we report that the adenovirus carrying a truncated FasL gene, named FasL minigene, encoding the full-length FasL protein (Ad-gFasL) is more efficient than the adenovirus carrying FasL cDNA (Ad-cFasL) in the induction of FasL expression in transduced cells. FasL minigene (2887 bp) lacking the second intron and a part of the 3′-UTR was created to reduce the gene length due to the size limitation of the adenoviral genome. The results show that, in transduced hepatocytes, strong expression of mRNA FasL appeared after 10 h for Ad-gFasL, while for Ad-cFasL, a faint expression appeared after 16 h. For Ad-gFasL, the protein expression was noticed starting with 0.5 transfection units (TU)/cell, while for Ad-cFasL, it could not be revealed. FasL-expressing endothelial cells induced apoptosis of A20 cells in co-culture experiments. FasL-expressing cells may be exploitable in various autoimmune diseases such as graft-versus-host disease, chronic colitis, and type I diabetes.

scholarly journals The Death Domain Kinase RIP Is Essential for TRAIL (Apo2L)-Induced Activation of IκB Kinase and c-Jun N-Terminal Kinase

2000 ◽  
Vol 20 (18) ◽  
pp. 6638-6645 ◽  
Author(s):  
Yong Lin ◽  
Anne Devin ◽  
Amy Cook ◽  
Maccon M. Keane ◽  
Michelle Kelliher ◽  
...  
Keyword(s):  
Ectopic Expression ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Death Domain ◽  
Iκb Kinase ◽  
Tnf Superfamily ◽  
Key Factor ◽  
Induced Apoptosis ◽  
Jnk Activation ◽  
Necrosis Factor

ABSTRACT Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) (Apo2 ligand [Apo2L]) is a member of the TNF superfamily and has been shown to have selective antitumor activity. Although it is known that TRAIL (Apo2L) induces apoptosis and activates NF-κB and Jun N-terminal kinase (JNK) through receptors such as TRAIL-R1 (DR4) and TRAIL-R2 (DR5), the components of its signaling cascade have not been well defined. In this report, we demonstrated that the death domain kinase RIP is essential for TRAIL-induced IκB kinase (IKK) and JNK activation. We found that ectopic expression of the dominant negative mutant RIP, RIP(559–671), blocks TRAIL-induced IKK and JNK activation. In the RIP null fibroblasts, TRAIL failed to activate IKK and only partially activated JNK. The endogenous RIP protein was detected by immunoprecipitation in the TRAIL-R1 complex after TRAIL treatment. More importantly, we found that RIP is not involved in TRAIL-induced apoptosis. In addition, we also demonstrated that the TNF receptor-associated factor 2 (TRAF2) plays little role in TRAIL-induced IKK activation although it is required for TRAIL-mediated JNK activation. These results indicated that the death domain kinase RIP, a key factor in TNF signaling, also plays a pivotal role in TRAIL-induced IKK and JNK activation.

Mechanisms for T-cell selective cytotoxicity of arabinosylguanine

Blood ◽  
2003 ◽  
Vol 102 (5) ◽  
pp. 1842-1848 ◽  
Author(s):  
Carlos O. Rodriguez ◽  
Christine M. Stellrecht ◽  
Varsha Gandhi
Keyword(s):  
T Cell ◽  
Cell Lines ◽  
Fas Ligand ◽  
Myeloid Cell ◽  
S Phase ◽  
Soluble Fas ◽  
Fasl Gene ◽  
Polymerase Chain ◽  
B Lineage ◽  
Induced Apoptosis

AbstractNelarabine, prodrug of arabinosylguanine (ara-G), has demonstrated T-lymphoblastic antileukemic activity in cell lines and in the clinic. To investigate the mechanism for lineage-specific toxicity, the effects of ara-G were compared in CEM (T-lymphoblast), Raji (B-lymphoblast), and ML-1 (myeloid) cell lines. CEM cells were the most sensitive to ara-G–induced apoptosis and accumulated the highest levels of ara-G triphosphate (ara-GTP). However, compared with myeloid and B-lineage cell lines, CEM cells incorporated fewer ara-G molecules—which were at internucleotide positions in all 3 cell lines— into DNA. Ara-G induced an S-phase arrest in both Raji and ML-1, while in CEM the S-phase cells decreased with a concomitant increase in the sub-G1 population. Within 3 hours of ara-G treatment, the levels of soluble Fas ligand (sFasL) in the medium increased significantly in CEM cultures. In parallel, an induction of FasL gene expression was observed by real-time reverse transcriptase–polymerase chain reaction (RT-PCR). Pretreatment of CEM cells with a Fas antagonistic antibody inhibited ara-G–mediated cell death. These results demonstrate that high ara-GTP accumulation in T cells results in an S phase–dependent apoptosis induced by ara-G incorporation into DNA, which may lead to a T cell–specific signal for the induction and liberation of sFasL. Subsequently, the sFasL induces an apoptotic response in neighboring non–S-phase cells. In contrast, myeloid and B cells accumulated lower levels of ara-GTP and arrested in S phase, blocking any apoptotic signaling.

scholarly journals Epstein–Barr virus-infected cells release Fas ligand in exosomal fractions and induce apoptosis in recipient cells via the extrinsic pathway

2015 ◽  
Vol 96 (12) ◽  
pp. 3646-3659 ◽  
Author(s):  
Waqar Ahmed ◽  
Pretty S. Philip ◽  
Samir Attoub ◽  
Gulfaraz Khan
Keyword(s):  
Epstein Barr Virus ◽  
Fas Ligand ◽  
Type I ◽  
Dependent Manner ◽  
Extrinsic Pathway ◽  
Barr Virus ◽  
Infected Cells ◽  
Epstein Barr ◽  
Induced Apoptosis ◽  
Cellular Components

Epstein–Barr virus (EBV; human herpesvirus 4) is an oncogenic herpesvirus implicated in the pathogenesis of several human malignancies. A number of recent studies indicate that EBV can manipulate the local microenvironment by excreting viral and cellular components in nanovesicles called exosomes. In this study, we investigated the impact of EBV-derived exosomes on apoptosis of recipient cells and the molecular pathway involved in this process. Exosomes from EBV-infected but not from non-infected cells induced apoptosis in a number of different cell types, including B-cells, T-cells and epithelial cells. However, this phenomenon was not universal and the Burkitt's lymphoma-derived B-cell line BJAB was found to be resistant to apoptosis. Exosomes from both type I and type III EBV latently infected cells induced apoptosis in a dose- and time-dependent manner. Moreover, cells exposed to EBV exosomes did not form colonies in soft agar assays. We further show that fluorescently labelled exosomes derived from EBV-infected cells are taken up by non-infected cells and induce apoptosis via the extrinsic pathway. Inhibition of caspase-3/7/8 blocks EBV exosome-mediated apoptosis. Furthermore, our data indicate that EBV exosomes trigger apoptosis through the Fas ligand (FasL)-mediated extrinsic pathway, as FasL was present in EBV exosomal fractions and anti-FasL antibodies could block EBV exosome-mediated apoptosis. Together, these data support the notion that EBV hijacks the exosome pathway to excrete viral and cellular components that can modulate its microenvironment.

scholarly journals FasL Is Required for Osseous Healing in Extraction Sockets in Mice

2021 ◽  
Vol 12 ◽  
Author(s):  
Karol Alí Apaza Alccayhuaman ◽  
Patrick Heimel ◽  
Jung-Seok Lee ◽  
Stefan Tangl ◽  
Franz J. Strauss ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Tooth Extraction ◽  
Fas Ligand ◽  
Healing Process ◽  
Apical Part ◽  
Trabecular Thickness ◽  
Tnf Superfamily ◽  
Significant Difference ◽  
Coronal Region ◽  
Necrosis Factor

Fas ligand (FasL) is a member of the tumor necrosis factor (TNF) superfamily involved in the activation of apoptosis. Assuming that apoptosis is initiated after tooth extraction it is reasonable to suggest that FasL may play a pivotal role in the healing of extraction sockets. Herein, we tested the hypothesis of whether the lack of FasL impairs the healing of extraction sockets. To this end, we extracted upper right incisors of FasL knockout (KO) mice and their wildtype (WT) littermates. After a healing period of two weeks, bone volume over total volume (BV/TV) via µCT and descriptive histological analyses were performed. µCT revealed that BV/TV in the coronal region of the socket amounted to 39.4% in WT and 21.8% in KO, with a significant difference between the groups (p=0.002). Likewise, in the middle region of the socket, BV/TV amounted to 50.3% in WT and 40.8% in KO (p<0.001). In the apical part, however, no difference was noticed. Consistently, WT mice displayed a significantly higher median trabecular thickness and a lower trabecular separation when compared to the KO group at the coronal and central region of the socket. There was the overall tendency that in both, female and male mice, FasL affects bone regeneration. Taken together, these findings suggest that FasL deficiency may reduce bone regeneration during the healing process of extraction sockets.

scholarly journals Keratin attenuates tumor necrosis factor–induced cytotoxicity through association with TRADD

2001 ◽  
Vol 155 (3) ◽  
pp. 415-426 ◽  
Author(s):  
Hiroyasu Inada ◽  
Ichiro Izawa ◽  
Miwako Nishizawa ◽  
Eriko Fujita ◽  
Tohru Kiyono ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Epithelial Cells ◽  
Tumor Necrosis ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Type I ◽  
Death Domain ◽  
Interacting Protein ◽  
Induced Apoptosis ◽  
Necrosis Factor

Keratin 8 and 18 (K8/18) are the major components of intermediate filament (IF) proteins of simple or single-layered epithelia. Recent data show that normal and malignant epithelial cells deficient in K8/18 are nearly 100 times more sensitive to tumor necrosis factor (TNF)–induced cell death. We have now identified human TNF receptor type 1 (TNFR1)–associated death domain protein (TRADD) to be the K18-interacting protein. Among IF proteins tested in two-hybrid systems, TRADD specifically bound K18 and K14, type I (acidic) keratins. The COOH-terminal region of TRADD interacted with the coil Ia of the rod domain of K18. Endogenous TRADD coimmunoprecipitated with K18, and colocalized with K8/18 filaments in human mammary epithelial cells. Overexpression of the NH2 terminus (amino acids 1–270) of K18 containing the TRADD-binding domain as well as overexpression of K8/18 in SW13 cells, which are devoid of keratins, rendered the cells more resistant to killing by TNF. We also showed that overexpressed NH2 termini of K18 and K8/18 were associated with endogenous TRADD in SW13 cells, resulting in the inhibition of caspase-8 activation. These results indicate that K18 may sequester TRADD to attenuate interactions between TRADD and activated TNFR1 and moderate TNF-induced apoptosis in simple epithelial cells.

Insulin-Induced Lipoatrophy in Type I Diabetes: A possible tumor necrosis factor- -mediated dedifferentiation of adipocytes

Diabetes Care ◽  
1996 ◽  
Vol 19 (11) ◽  
pp. 1283-1285 ◽  
Author(s):  
C. Atlan-Gepner ◽  
P. Bongrand ◽  
C. Farnarier ◽  
L. Xerri ◽  
R. Choux ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Tumor Necrosis ◽  
Type I Diabetes ◽  
Type I ◽  
Necrosis Factor

scholarly journals Complete mutation scan of the human Fas ligand gene: Linkage studies in Type I diabetes mellitus families

Diabetologia ◽  
2002 ◽  
Vol 45 (1) ◽  
pp. 134-139 ◽  
Author(s):  
R. L. Nolsøe ◽  
O. P. Kristiansen ◽  
Z. M. Larsen ◽  
J. Johannesen ◽  
F. Pociot ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Fas Ligand ◽  
Type I Diabetes ◽  
Type I Diabetes Mellitus ◽  
Type I ◽  
Linkage Studies ◽  
Gene Linkage ◽  
Human Fas Ligand

Alginate Polylysine Microcapsules as Immune Barrier: Permeability of Cytokines and Immunoglobulins over the Capsule Membrane

1997 ◽  
Vol 6 (4) ◽  
pp. 387-394 ◽  
Author(s):  
Bård Kulseng ◽  
Beate Thu ◽  
Terje Espevik ◽  
Gudmund Skjåk-Bræk
Keyword(s):  
Tumor Necrosis Factor ◽  
Pancreatic Islets ◽  
Membrane Permeability ◽  
Acid Content ◽  
Tumor Necrosis ◽  
Type I Diabetes ◽  
Type I ◽  
Mannuronic Acid ◽  
Mouse Islets ◽  
Necrosis Factor

Transplantation of pancreatic islets in alginate polylysine microcapsules is a potential useful method for treating type I diabetes. In this study, the permeability for alginate-polylysine microcapsules to cytokines an immunoglobulines has been investigated by a newly developed method. Magnetic monodisperse polymer particles (Dynabeads) coated with antibodies against selected proteins were encapsulated in 0.7 mm alginate polylysine microcapsules. The capsule membrane permeability to IgG (150 kDa). Transferrin (81 kDa), Tumor necrosis factor (TNF, 51 kDa), Interleukin-1β (IL-1β, 17.5 kDa), and insulin (5.8 kDa) was estimated by measuring the binding of 125I-labeled proteins to the encapsulated antibody coated Dynabeads. Capsules with an inhomogeneous solid gel core were made of alginates with high guluronic or high mannuronic acid content and poly-l (PLL)- or poly-d-lysine (PDL) of concentrations varied from 0.05-0.2%. The various capsules examined were all impermeable to IgG. The capsules made with a PLL-, but not PDL-membranes were permeable for transferrin. IL-1β was found to penetrate all of the different capsule types. The high-G capsules, however, could be made impermeable to TNF and still allowed transferrin to pass. The permeability of these capsules to IL-1β, but not to TNF was confirmed in an assay where mouse islets of Langerhans were incubated with TNF and IL-1β, and comparing the IL-6 for encapsulated and non-encapsulated islets.

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