Ridostin Induces Transcription of a Wide Spectrum of Interferon Genes in Human Cells

Соколова, Т. М.; Шувалов, А.Н.; Telkov, M. V.; Kolodyazhnaya, Lyubov Vladimirovna; Ершов, Ф. И.

doi:10.1007/s10517-013-2313-z

Cited by 3 publications

(5 citation statements)

References 9 publications

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“…The validation of our findings by genorm largely corroborated the findings from the normfinder analysis; however, normfinder allows analysing the stability in relation to different groups and suggests alternative reference genes for analyses involving RRMS‐IFN‐ β patients. This finding is consistent with the marked effects of interferon‐ β treatment on gene expression and previous findings of effects of interferon signalling on the stability of reference genes . Manoli et al .…”

Section: Discussionsupporting

confidence: 92%

“…However, inclusion of RRMS-IFN-b patients in the analyses increased the stability value, that is creating a lower stability of the reference gene. This indicates that interferon-b treatment increases the variation in reference gene expression, which is in accordance with previous observations and emphasizes the need for reference gene validation [21,22]. When analysing stability in relation to cell subsets (Analyses 3-5), we observed a general marked increase in the stability values, indicating that cell subsets contribute to substantial variation in the expression of reference genes.…”

Section: Discussionsupporting

confidence: 91%

“…The validation of our findings by GENORM largely corroborated the findings from the NORMFINDER analysis; however, NORMFINDER allows analysing the stability in relation to different groups and suggests alternative reference genes for analyses involving RRMS-IFN-b patients. This finding is consistent with the marked effects of interferon-b treatment on gene expression and previous findings of effects of interferon signalling on the stability of reference genes [21,22]. Manoli et al [27] applied NORMFINDER alongside with two other algorithms, BestKeeper and GENORM, and found that there were no significant differences in the results when applying all three algorithms on their data.…”

Section: Cells and Non-b Cells)supporting

confidence: 86%

“…Additionally, studies of multiple sclerosis immunopathogenesis often involve studies of peripheral blood mononuclear cell (PBMC) subsets, but only few studies have examined the stability of reference genes across the different leucocyte subsets and these have demonstrated marked variability in reference gene expression levels in varying subsets . Furthermore, interferon signalling is known to affect reference gene expression and interferon‐ β (IFN‐ β ) treatment to induce a plethora of genes, and no studies have validated reference genes for interferon treatment .…”

Section: Introductionmentioning

confidence: 99%

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Identification of Suitable Reference Genes for Peripheral Blood Mononuclear Cell Subset Studies in Multiple Sclerosis

et al. 2015

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Quantitative real-time PCR (qPCR) involves the need of a proper standard for normalizing the gene expression data. Different studies have shown the validity of reference genes to vary greatly depending on tissue, cell subsets and experimental context. This study aimed at the identification of suitable reference genes for qPCR studies using different peripheral blood cell subsets (whole blood (WB) cells, peripheral blood mononuclear cells (PBMCs) and PBMC subsets (CD4 + T cells, CD8 + T cells, NK cells, monocytes, B cells and dendritic cells) from healthy controls (HC), patients with relapsing-remitting multiple sclerosis (RRMS) and interferon-b-treated patients with RRMS (RRMS-IFN-b). Eight candidate reference genes (CASC3, EEF1A1, GAPDH, HPRT1, RPLP0, UBC, UBE2D2 and YWHAZ) were analysed using NORMFINDER and GENORM algorithms to identify the most stably expressed genes. We found reference gene expression varied most across cell subsets, and less variation between the donor groups. UBE2D2 was the most stably expressed gene across both HC and RRMS patients and across cell subsets, while UBC was the most stably expressed gene between HC, RRMS and RRMS-IFN-b patients. UBE2D2 and HPRT1 was the most stable combination for analyses of cell subsets between HC and RRMS patients, while the combination of UBC and YWHAZ was superior for analysis of cell subsets between HC, RRMS and RRMS-IFN-b groups. GAPDH was generally unsuitable for blood cell subset studies in multiple sclerosis. In conclusion, we found that blood cell subsets result in marked variation in reference gene expression, and we identified suitable reference genes for studies involving PBMC subsets, RRMS patients and interferon-b treatment.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 91%

Section: Cells and Non-b Cells)supporting

confidence: 86%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Identification of Suitable Reference Genes for Peripheral Blood Mononuclear Cell Subset Studies in Multiple Sclerosis

et al. 2015

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“…Таким образом, при введении в организм дсРНК стимулирует образование эндогенных ИФН-α, ИФН-β и ИФН-γ, важнейших цитокинов иммунного ответа, которые индуцируют дифференцировку миелоидных клеток, стимулируют фагоцитоз нейтрофилов и макрофагов, активируют натуральные киллеры, усиливают Т-хелперный ответ Тh1-типа и таким образом запускают врожденный и адаптивный иммунный ответ. Влияние ридостина как препарата индуктора ИФН на транскрипцию генов семейства ИФН в фибробластах и лимфоцитах человека было изучено ранее с помощью количественной ПЦР-РВ, что проявилось в увеличении транскрипции генов ИФН-β, олигоаденилатсинтетазы и дсРНК-зависимой протеинкиназы [30].…”

Section: время чunclassified

Efficacy of interferon inducers against Chikungunya virus <i>in vitro</i>

Otrashevskaja,

Kaa,

Samartseva

et al. 2023

Biological Products. Prevention, Diagnosis, Treatment

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Scientific relevance. To date, no specific antivirals have been approved to treat and prevent Chikungunya fever, its complications, and sequelae. Therefore, the development of therapeutic and preventive medicinal products against Chikungunya virus (CHIKV), including interferon inducers, is gaining relevance.Aim. The authors aimed to study the effectiveness of prophylactic administration of an interferon inducer against CHIKV in an in vitro model.Materials and methods. The study used two cell lines (Vero and А549), a CHIKV strain (Nika2021), and an interferon-inducing medicinal product (double-stranded RNA sodium salt) at two doses (250 μg/mL and 500 μg/mL) administered at two schedules: Prevention (4 h prior to the virus challenge) and Emergency Prevention (at the time of the virus challenge). The authors determined the CHIKV titre by its cytopathogenic effect, the CHIKV RNA content by the cycle threshold value in real-time reverse-transcription polymerase chain reaction, and the concentration of cytokines using the enzyme immunoassay method. The study monitored the changes in CHIKV biological activity, CHIKV RNA levels, and the production of interferon-alpha (IFN-α), interferon-gamma (IFN-γ), interleukin-6 (IL-6), and tumour necrosis factor-alpha (TNF-α) in cells over time. The statistical analysis of the resulting data used Microsoft Office Excel 2016 and StatTech.Results. The medicinal product at doses of 250 μg/mL and 500 μg/mL stimulated the production of both IFN-α and IFN-γ (IFN-α to a greater extent than IFN-γ) in both cell lines (in A549 to a greater extent than in Vero). The changes in CHIKV RNA levels with time corresponded to those of the virus titre. In general, CHIKV RNA levels in Vero cells were significantly higher than those in A549 cells (р<0.002 at 250 μg/mL and р<0.0005 at 500 μg/mL). The CHIKV RNA content after preventive interferon inducer administration was significantly lower than that in the control experiment (challenge without administration of the medicinal product) for both doses and both cell lines (р<0.002 for Vero cells; р<0.0003 for А549 cells). The CHIKV RNA content after interferon inducer administration as emergency prevention was significantly lower than that in the control experiment (р<0.05 for Vero cells; р<0.003 for А549 cells). The study demonstrated the efficacy of the interferon inducer against CHIKV and a higher applicability of the A549 cell line to studying antiviral activity in vitro. The authors observed the production of IL-6 and TNF-α by intact cells of both lines.Conclusions. According to the results, the studied interferon inducer has a positive antiviral effect against CHIKV in vitro, with the antiviral effect degree depending on the cell line used. This experimental study demonstrated the need to carefully select the cell line for a study in accordance with its objectives and to evaluate the production of cytokines by a monolayer of cells before stimulation with viruses and/or medicinal products.

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Stimulating effect of double-stranded yeast RNA on the activity of interferon system genes

Батенева¹,

Гамалей²,

Lebedev³

et al. 2021

Med. immunol.

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Influence of double-stranded RNA (dsRNA) from Saccharomyces cerevisiae yeast upon expression levels of the macrophage genes encoding TLR3 receptor, interferons alpha and beta (IFNα, IFNβ), 2’,5’-oligoadenylate synthetase (OAS) and protein kinase R (PKR) enzymes has been studied in the J774 mouse histiocytic cell culture and in vivo in Balb/c mice. It has been shown that dsRNA exerts a selective activating effect on genes of TLR3 receptor, antiviral proteins IFNα, IFNβ, and OAS, both in vitro and in vivo. With J774 cell culture, the highest induction capacity was observed for the IFNβ gene: 365 to 802-fold. The stimulatory effect was dependent on the dose of dsRNA in the range of 16.9 to 125 μg/ml. The preparation enhanced IFNα gene activity to lesser degree (more than 10-fold), TLR3 and OAS (3 to 4-fold), while the expression levels for these genes were not significantly dependent on the dose of dsRNA. The stimulating effect of dsRNA was dosedependent in murine peritoneal macrophages. The maximum activating effect of the preparation was shown upon administration of the effective antiviral dose (0.5 mg of dsRNA/kg). Five hours after intraperitoneal injection of dsRNA, the highest level of mRNA synthesis was observed for IFNα (54-fold), OAS (43-fold) and TLR3 (28-fold) genes. Expression of the IFNβ gene increased to a lesser degree (9-fold). An increase in the dose of preparation to 1.5 mg/kg led to decrease of the stimulatory effect. Expression levels of the IFNα, TLR3, and OAS genes in that case decreased by 2-4-fold as compared to a lower dose, and the PKR gene expression was 5-fold lower compared to the control. One day after dsRNA administration, a tendency was observed for both experimental groups towards a decreased transcription of macrophage genes, if compared with the 5-hour term. The weakening of gene activity was less pronounced in animals treated with dsRNA at the dose of 1.5 mg/kg. The transcription indices for IFNβ, OAS, and TLR3 genes were much higher during this period (5-10-fold higher than the control values). The dynamics of PKR gene transcription in both experimental systems was significantly different from the expression of other studied genes. The dsRNA preparation at this dose range did not have a pronounced stimulatory effect upon expression of this gene. A moderate increase in PKR gene activity in macrophages of mice was observed only a day following intraperitoneal administration of dsRNA. Concentrations and length of dsRNA molecules are known to be critical factors to the PKR gene activation. An ability to increase the expression of the gene is shown at low dsRNA concentrations (10-7 g/ml and below), while highly polymeric dsRNAs weaken the gene activity. Since the doses and concentrations of dsRNA used in our experiments were significantly different from those mentioned above, it could, in general, affect regulation of PKR gene transcription towards reduction of the stimulatory effect.

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Ridostin Induces Transcription of a Wide Spectrum of Interferon Genes in Human Cells

Cited by 3 publications

References 9 publications

Identification of Suitable Reference Genes for Peripheral Blood Mononuclear Cell Subset Studies in Multiple Sclerosis

Identification of Suitable Reference Genes for Peripheral Blood Mononuclear Cell Subset Studies in Multiple Sclerosis

Efficacy of interferon inducers against Chikungunya virus <i>in vitro</i>

Stimulating effect of double-stranded yeast RNA on the activity of interferon system genes

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