Exogenous allogenic fragmented double-stranded DNA is internalized into human dendritic cells and enhances their allostimulatory activity

Alyamkina, E. A.; Долгова, Е. В.; Likhacheva, Anastasia S.; Рогачев, В. А.; Sebeleva, Tamara E.; Николин, В. П.; Попова, Н. А.; Киселева, Елена; Orishchenko, Кonstantin E.; Сахно, Л. В.; Gel’fgat, Evgeniy L.; Останин, А. А.; Черных, Е. Р.; Zagrebelniy, Stanislav N.; Богачев, С. С.; Shurdov, Mikhail A.

doi:10.1016/j.cellimm.2010.01.005

Cited by 18 publications

(25 citation statements)

References 41 publications

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“…Proceeding on reported observations [14,39,47], we assumed that this inhibition may be due to the inducer effect of dsDNA on DC maturation in vivo that causes effective presentation of antigens of tumor lysate and activates antitumor mechanisms of the adaptive immunity.…”

Section: Resultsmentioning

confidence: 99%

“…Dendritic cells (DCs), which are capable of activating T-lymphocytes, including naive T-cells, have an important role in triggering and development of the adaptive immunity [9,13,14]. Mature DCs that express MHC antigens of class I and class II, also the various costimulatory molecules CD40, CD54, CD80, and CD86 are capable of only presenting foreign antigens within the MHC complex [15-21].…”

Section: Introductionmentioning

confidence: 99%

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A strategy of tumor treatment in mice with doxorubicin-cyclophosphamide combination based on dendritic cell activation by human double-stranded DNA preparation

Alyamkina

Николин

Попова

et al. 2010

Genet Vaccines Ther

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BackgroundImmunization of mice with tumor homogenate after combined treatment with cyclophosphamide (CP) and double-stranded DNA (dsDNA) preparation is effective at inhibition of growth of tumor challenged after the treatment. It was assumed that this inhibition might be due to activation of the antigen-presenting cells. The purpose was to develop improved antitumor strategy using mice. We studied the combined action of cytostatics doxorubicin (Dox) plus CP with subsequent dsDNA preparation on tumor growth.MethodsThree-month old CBA/Lac mice were used in the experiments. Mice were injected with CP and human dsDNA preparation. The percentage of mature dendritic cells (DCs) was estimated by staining of mononuclear cells isolated from spleen and bone marrow 3, 6, and 9 days later with monoclonal antibodies CD34, CD80, and CD86. In the next set of experiments, mice were given intramuscularly injections of 1-3 × 105 tumor cells. Four days later, they were injected intravenously with 6-6.7 mg/kg Dox and intraperitoneally with 100-200 mg/kg CP; 200 mkg human DNA was injected intraperitoneally after CP administration. Differences in tumor size between groups were analyzed for statistical significance by Student's t-test. The MTT-test was done to determine the cytotoxic index of mouse leucocytes from treated groups.ResultsThe conducted experiments showed that combined treatment with CP and dsDNA preparation produce an increase in the total amount of mature DCs in vivo. Treatment of tumor bearers with preparation of fragmented dsDNA on the background of pretreatment with Dox plus CP demonstrated a strong suppression of tumor growth in two models. RLS, a weakly immunogenic, resistant to alkalyting cytostatics tumor, grew 3.4-fold slower when compared with the control (p < 0.001). In experiment with Krebs-2 tumor, only 2 of the 10 mice in the Dox+CP+DNA group had a palpable tumor on day 16. The cytotoxic index of leucocytes was 86.5% in the Dox+CP+DNA group, but it was 0% in the Dox+CP group.ConclusionsThus, the set of experiments we performed showed that exogenous dsDNA, when administered on the background of pretreatment with Dox plus CP, has an antitumor effect possibly due to DC activation.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A strategy of tumor treatment in mice with doxorubicin-cyclophosphamide combination based on dendritic cell activation by human double-stranded DNA preparation

Alyamkina

Николин

Попова

et al. 2010

Genet Vaccines Ther

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“…Human DNA was isolated from placentas of healthy women using a phenol-free method, and sonicated as described in (Alyamkina et al, 2010).…”

Section: Administering the Cp And Human Dna Preparationmentioning

confidence: 99%

“Delayed death” phenomenon: A synergistic action of cyclophosphamide and exogenous DNA

Долгова¹,

Proskurina²,

Николин³

et al. 2012

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“…Дендритные клетки были генерированы из моноци тов крови трех доноров в стандартных условиях с использо-ванием ГМ-КСФ и ИФН-α (Alyamkina et al, 2010a). Полу-ченные ДК были обработаны следующим образом: 1) кон-троль -интактные ДК; 2) ДК, обработанные хлорохи-ном (100 мкM); 3) ДК + двуцепочечная ДНК (10 мкг/ мл); 4) ДК, обработанные хлорохином (100 мкM), плюс дву-цепочечная ДНК (10 мкг/мл).…”

Section: результатыunclassified

“…В наших исследованиях было показано, что двуцепо-чечная геномная self ДНК естественным путем (т. е. без применения каких-либо трансфецирующих агентов) эф-фек тивно активирует ДК человека миелоидного типа, генерированные ex vivo из моноцитов крови, что проявля-ется усилением экспрессии активационных и дифферен-цировочных антигенов, аллостимуляторной активности ДК в смешанной культуре лимфоцитов и способности ДК индуцировать пролиферацию перфоринсодержащих ци-тотоксических Т-лимфоцитов (Alyamkina et al, 2010a(Alyamkina et al, , b, 2012Orishchenko et al, 2013). Было показано также, что препарат двуцепочечной ДНК человека стимулирует продукцию целого ряда цитокинов и хемокинов монону-клеарными клетками цельной крови: ИЛ-1β, ИЛ-6, ИЛ-8, ФНО-α, ИФН-α, ИФН-γ, MCP1, VEGF, ИЛ-1РА и ИЛ-10 ( Orishchenko et al, 2013).…”

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Expression of genes of cytokines, transcription factors and differentiation antigens in human dendritic cells activated by double-stranded DNA

Proskurina¹,

Orishchenko²,

Potter³

et al. 2017

Vestn. VOGiS

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One of the most important properties of extracellular double-stranded DNA related to the treatment of various diseases is its ability to activate effector cells of the immune system (anti-tumor and vaccinal immunity) through dendritic cells (DCs). The stimulatory effect of DNA on DCs is mediated by the TLR9 signaling pathway and/or through a system of cytosolic sensors and is manifested by increased expression of MHC class II antigens and costimulatory molecules and by increased synthesis of immunoregulatory cytokines. In this work, the expression of cytokines, differentiation antigens and transcription factor genes has been investigated in DCs activated by double-stranded human DNA (i) without any additional factors, (ii) using a lipophilic agent, and (iii) by blocking TLR9 with chloroquine. Evaluation of the DNA effect was carried out after the 6-and 24-hour exposure. It was found that the preparation of double-stranded DNA transfected by Lipofectamine 2000 boosts DCs at the same level as Poly(dA : dT), a synthetic equivalent of double-stranded DNA. It was discovered that combined application of DNA and chloroquine enhances expression of the IFN-α, IL8, МСР1, VEGF, CD25, and CD83 genes by hour 24 of incubation. It was for the first time shown that genomic "self" double-stranded DNA as a mono agent activates mRNA synthesis of cytokines IFN-α, IFN-β, IFN-γ, IL8, IL10, and VEGF in DCs at 6 hours of induction.Key words: chloroquine; double-stranded DNA; cytokines; dendritic cells; real-time PCR.Одно из важнейших свойств экстраклеточной двуцепочечной ДНК, имеющее отношение к терапии различных заболеваний, -это ее способность через дендритные клетки (ДК) активировать эффекторные клетки иммунной системы (противоопухолевый и вакцинальный иммунитет). Стимулирующий эффект ДНК на ДК опосредуется через TLR9 сигнальный путь и/или через систему цитозольных сенсоров и проявляется усилением экспрессии МНС антигенов II класса и костимулирующих молекул, а также синтеза иммунорегуляторных цитокинов. В настоящей работе была иссле-дована экспрессия генов цитокинов, антигенов дифференцировки и факторов транскрипции в ДК, активированных двуцепочечной ДНК человека: (1) без использования каких-либо дополнительных факторов, (2) при использовании липофильного агента и (3) при блокировании TLR9 хлорохином. Оценка эффекта ДНК проводи-лась после 6-и 24-часовой экспозиции. Показано, что препарат двуцепочечной ДНК при трансфекции липофектамином активиру-ет ДК на таком же уровне, как и Poly(dA : dT), синтетический аналог двуцепочечной ДНК. Установлено, что совместная обработка ДНК и хлорохином усиливает экспрессию генов ИФН-α, ИФН-β, ИФН-γ, ИЛ8, МСР1, VEGF, CD25 и CD83 к 24 ч инкубации. Впервые показано, что геномная «self» двуцепочечная ДНК как монопрепарат акти-вирует синтез мРНК ИФН-α, ИФН-β, ИФН-γ, ИЛ8, ИЛ10 и VEGF в ДК к 6 часам индукции.

show abstract

Exogenous allogenic fragmented double-stranded DNA is internalized into human dendritic cells and enhances their allostimulatory activity

Cited by 18 publications

References 41 publications

A strategy of tumor treatment in mice with doxorubicin-cyclophosphamide combination based on dendritic cell activation by human double-stranded DNA preparation

A strategy of tumor treatment in mice with doxorubicin-cyclophosphamide combination based on dendritic cell activation by human double-stranded DNA preparation

“Delayed death” phenomenon: A synergistic action of cyclophosphamide and exogenous DNA

Expression of genes of cytokines, transcription factors and differentiation antigens in human dendritic cells activated by double-stranded DNA

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