Recombinant TNF-binding protein from variola virus as a novel potential TNF antagonist

Gileva, I. P.; Nepomnyashchikh, T. S.; Ryazankin, I. A.; Щелкунов, С. Н.

doi:10.1134/s0006297909120098

Cited by 6 publications

(5 citation statements)

References 26 publications

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“… 42 , 43 Similarly, VAR F1L may be a valid target for development of antiviral therapeutics. Current strategies for the development of antiviral therapeutics against VAR have relied mostly on the use of homologous target molecules from VV, monkeypox or ectromelia virus, with only a small number of studies utilizing VAR-encoded proteins, 44 , 45 , 46 and even fewer studies using their structures. 47 , 48 The marked differences between VAR and VV F1L suggest that studies of VAR proteins may provide unexpected avenues for therapeutic intervention that may not be predicted based on available data from the closely related and much better understood VV.…”

Section: Discussionmentioning

confidence: 99%

Variola virus F1L is a Bcl-2-like protein that unlike its vaccinia virus counterpart inhibits apoptosis independent of Bim

et al. 2015

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Subversion of host cell apoptosis is an important survival strategy for viruses to ensure their own proliferation and survival. Certain viruses express proteins homologous in sequence, structure and function to mammalian pro-survival B-cell lymphoma 2 (Bcl-2) proteins, which prevent rapid clearance of infected host cells. In vaccinia virus (VV), the virulence factor F1L was shown to be a potent inhibitor of apoptosis that functions primarily be engaging pro-apoptotic Bim. Variola virus (VAR), the causative agent of smallpox, harbors a homolog of F1L of unknown function. We show that VAR F1L is a potent inhibitor of apoptosis, and unlike all other characterized anti-apoptotic Bcl-2 family members lacks affinity for the Bim Bcl-2 homology 3 (BH3) domain. Instead, VAR F1L engages Bid BH3 as well as Bak and Bax BH3 domains. Unlike its VV homolog, variola F1L only protects against Bax-mediated apoptosis in cellular assays. Crystal structures of variola F1L bound to Bid and Bak BH3 domains reveal that variola F1L forms a domain-swapped Bcl-2 fold, which accommodates Bid and Bak BH3 in the canonical Bcl-2-binding groove, in a manner similar to VV F1L. Despite the observed conservation of structure and sequence, variola F1L inhibits apoptosis using a startlingly different mechanism compared with its VV counterpart. Our results suggest that unlike during VV infection, Bim neutralization may not be required during VAR infection. As molecular determinants for the human-specific tropism of VAR remain essentially unknown, identification of a different mechanism of action and utilization of host factors used by a VAR virulence factor compared with its VV homolog suggest that studying VAR directly may be essential to understand its unique tropism.

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Section: Discussionmentioning

confidence: 99%

Variola virus F1L is a Bcl-2-like protein that unlike its vaccinia virus counterpart inhibits apoptosis independent of Bim

et al. 2015

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“…It should be noted that TNF BD, which was synthesized in E. coli cells, was not glycosy lated, whereas VARV CrmB, which was produced in the insect cells, was glycosylated [11].…”

Section: Tnf Binding Domain Of the Variola Virus Crmb Protein Synthesmentioning

confidence: 99%

TNF-Binding domain of the variola virus CrmB protein synthesized in Escherichia coli cells effectively interacts with human TNF

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Shekhovtsov

Nepomnyashchikh

et al. 2015

Dokl Biochem Biophys

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“…Интактным самкам (C57BL/6xDBA/2)F1 внутрибрюшинно однократно вводили ЛПС E. coli штамма 111: B4 (Sigma), в дозах 10 нг, 1 мкг и 100 мкг на мышь в PBS (PBS-10 мМ фосфатный буфер рH 7,4, 0,15 М NaCl), контрольной группе вводился соответствующий объем буфера. В качестве ингибитора TNFα был использован TNFα-связывающий домен белка CRMB виру-са натуральной оспы (TNF-BD) любезно предоставленный Щелкуновым С.Н., обозначаемый нами как TNFα-связывающий белок [3]. TNFαсвязывающий белок вводили за 30 мин.…”

Section: материалы и методыunclassified

Dynamics of cell-free DNA levels in the in vivo LPS-induced inflammation model

Демченко¹,

Гаврилова²,

Гойман³

et al. 2022

Russian Journal of Immunology

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An increased concentration of extracellular cell free DNA (cfDNA) is a distinctive characteristic of pathologies that mainly occur in acute inflammation (myocardial infarction, sepsis, stroke, trauma). The increase of cfDNA in chronic inflammatory processes, oncological, autoimmune diseases is less significant and is mainly due to aberrant cell death processes. One of such diseases is systemic lupus erythematosus (SLE). It has recently been shown that, in addition to increased cfDNA concentration, the degree of inflammation can reflect the N/L index (neutrophil to lymphocyte ratio), being a simple and informative marker of disease activity in patients with SLE. The aim of the study was to study the dynamics of the level of cfDNA and the N/L index in the model of LPS-induced inflammatory response as observed in intact mice, and their relation to the phenotypic heterogeneity of model SLE. We used female hybrid mice (C57Bl/6xDBA/2) F1 and female DBA/2 mice at the age of 6-8 weeks. LPS of E. coli strain 111: B4 (Sigma) was injected intraperitoneally once at doses of 10 ng, 1 g and 100 g per mouse in PBS. The control group was injected with the appropriate volume of buffer. The TNF-binding domain of the variola virus CRMB protein was used as an inhibitor of TNF, which was administered 30 min before the introduction of LPS. The dynamics of the response to LPS was assessed after 4, 8, 11, 24 hours by the N/L index and the level of cfDNA; at the zero point, the parameters were determined before the introduction of LPS. A day after a single injection of LPS at a dose of 1 g/mouse, a SLE model was induced on the same hybrid mice (double intravenous administration with an interval of 6 days of spleen cells of the DBA/2 line, 60-70 106 cells each). Three months later, with proteinuria of 3 mg/ mL or more, mice were assigned to the SLEnephritis+ group, with a protein of less than 3 mg/mL, to the SLEnephritis- group. Statistical processing of the results was carried out by nonparametric statistics using the MannWhitney test. Differences were considered statistically significant at p 0.05. It was found that the change in the N/L index, as well as the change in the level of cfDNA, depends on the dose of LPS administered. It was shown that the level of cfDNA reaches its maximum after 8 and 11 hours after the introduction of LPS is reliably reduced when using the inhibitor TNF. A retrospective analysis indicates that there is a definite relationship between the response of intact mice to LPS before induction of cGVHD, and their subsequent division into variants of SLEnephritis + and SLEnephritis - in the course of disease development.

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Recombinant TNF-binding protein from variola virus as a novel potential TNF antagonist

Cited by 6 publications

References 26 publications

Variola virus F1L is a Bcl-2-like protein that unlike its vaccinia virus counterpart inhibits apoptosis independent of Bim

Variola virus F1L is a Bcl-2-like protein that unlike its vaccinia virus counterpart inhibits apoptosis independent of Bim

TNF-Binding domain of the variola virus CrmB protein synthesized in Escherichia coli cells effectively interacts with human TNF

Dynamics of cell-free DNA levels in the in vivo LPS-induced inflammation model

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