Cell-free DNA in plasma as an essential immune system regulator

Korabečná, Marie; Zinková, Alžběta; Brynychova, Iva; Chylíková, Blanka; Přikryl, Petr; Šedová, Lucie; Neužil, Pavel; Šeda, Ondřej

doi:10.1038/s41598-020-74288-2

Cited by 28 publications

(25 citation statements)

References 61 publications

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“…4 and 5 ), showing no significant differences from the PRE cfDNA values (Supplementary Tables S5 and S6 ). Since elevated cfDNA concentrations have been discussed to possibly trigger enhanced inflammation 52 , or the production of anti-ds-DNA antibodies 5 , low increases and rapid decreases are positive aspects of cfDNA kinetics in SLE patients. With the established time and cost efficient assay, we determined the cfDNA kinetics in exercising SLE patients for the first time, underlining that exercise does not have a negative impact on the levels of cfDNA.…”

Section: Discussionmentioning

confidence: 99%

Validating quantitative PCR assays for cfDNA detection without DNA extraction in exercising SLE patients

Neuberger

Brahmer

Ehlert

et al. 2021

Sci Rep

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Circulating cell-free DNA (cfDNA) has been investigated as a screening tool for many diseases. To avoid expensive and time-consuming DNA isolation, direct quantification PCR assays can be established. However, rigorous validation is required to provide reliable data in the clinical and non-clinical context. Considering the International Organization for Standardization, as well as bioanalytical method validation guidelines, we provide a comprehensive procedure to validate assays for cfDNA quantification from blood plasma without DNA isolation. A 90 and 222 bp assay was validated to study the kinetics of cfDNA after exercise in patients with systemic lupus erythematosus (SLE). The assays showed ultra-low limit of quantification (LOQ) with 0.47 and 0.69 ng/ml, repeatability ≤ 11.6% (95% CI 8.1–20.3), and intermediate precision ≤ 12.1% (95% CI 9.2–17.7). Incurred sample reanalysis confirmed the precision of the procedure. The additional consideration of pre-analytical factors shows that centrifugation speed and temperature do not change cfDNA concentrations. In SLE patients cfDNA increases ~ twofold after a walking exercise, normalizing after 60 min of rest. The established assays allow reliable and cost-efficient quantification of cfDNA in minute amounts of plasma in the clinical setting. Additionally, the assay can be used as a tool to determine the impact of pre-analytical factors and validate cfDNA quantity and quality of isolated samples.

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Section: Discussionmentioning

confidence: 99%

Validating quantitative PCR assays for cfDNA detection without DNA extraction in exercising SLE patients

Neuberger

Brahmer

Ehlert

et al. 2021

Sci Rep

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show abstract

“…On the other hand, ecDNA is present in the circulation of healthy individuals and could have physiological roles that are yet unexplored. An interesting hypothesis is that ecDNA could act as an inflammation regulator under physiological conditions (50). It is, however, clear that ecDNA increases in systemic inflammation (51,52) and at least some of its fractions induce inflammation as well (53,54).…”

Section: Discussionmentioning

confidence: 99%

Circulating extracellular DNA is in association with continuous metabolic syndrome score in healthy adolescents

Celec

Janovičová

Gurecká

et al. 2021

Physiological Genomics

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Obesity is associated with chronic low-grade inflammation that eventually leads to metabolic complications. Extracellular DNA (ecDNA) is a damage-associated molecular pattern. Extracellular mitochondrial DNA can activate innate immunity. We hypothesized that ecDNA, especially of mitochondrial origin could be associated with components of the metabolic syndrome in young healthy probands. In a cross-sectional study healthy adolescents (n=1249) provided blood samples. Anthropometric data, blood pressure and blood counts were assessed. In addition, biochemical analysis of sera or plasma was conducted including the quantification of advanced oxidation protein products (AOPP) as a marker of oxidative stress induced by neutrophil or monocyte activation. Plasma ecDNA was isolated and measured using fluorometry. Nuclear and mitochondrial DNA were quantified using real time PCR. Males had higher total plasma ecDNA (15 (11-21) vs 11 (8-17) ng/ml; median (IQR)), nuclear (1760 (956-3273) vs 1153 (600-2292) GE/ml) and mitochondrial DNA (37181 (14836-90896) vs 30089 (12587-72286) GE/ml). EcDNA correlated positively with the continuous metabolic syndrome score (r= 0.158 for males and r= 0.134 for females). Stronger correlations were found between ecDNA of mitochondrial origin and AOPP (r= 0.202 and 0.186 for males and females respectively). Multivariate regression analysis revealed associations of nuclear DNA with leukocyte and erythrocyte counts. The results of this study on healthy adolescents show that circulating ecDNA is associated with the risk of metabolic syndrome, not with obesity per se. The association between mitochondrial ecDNA and AOPP requires further attention as it supports a potential role of mitochondria-induced sterile inflammation in the pathogenesis of the metabolic syndrome.

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“…In the lupus model, we followed the levels of cfDNA in detail during the first 10 days after pristane injection. On day 10, A 2A R levels were low in A 1 R-KO mice compared to WT mice while cfDNA levels were significantly higher than the WT mice, which might contribute to the severe development of the disease in this group (63).…”

Section: Discussionmentioning

confidence: 86%

A1 and A2A adenosine receptors play a protective role to reduce prevalence of autoimmunity following tissue damage

Riff

Naamani

Mazar

et al. 2021

Clinical &Amp; Experimental Immunology

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Adenosine is a potent modulator that has a tremendous effect on the immune system.Adenosine affects T cell activity, and is necessary in maintaining the T helper/regulatory T cell (T reg ) ratio. Adenosine signalling is also involved in activating neutrophils and the formation of neutrophil extracellular traps (NETs), which has been linked to autoimmune disorders. Therefore, adenosine, through its receptors, is extremely important in maintaining homeostasis and involved in the development of autoimmune diseases. In this study, we aim to evaluate the role of adenosine A 1 and A 2A receptors in involvement of autoimmune diseases. We studied adenosine regulation by NETosis in vitro, and used two murine models of autoimmune diseases: type I diabetes mellitus (T1DM) induced by low-dose streptozotocin and pristane-induced systemic lupus erythematosus (SLE). We have found that A 1 R enhances and A 2A R suppresses NETosis. In addition, in both models, A 1 R-knock-out (KO) mice were predisposed to the development of autoimmunity.In the SLE model in wild-type (WT) mice we observed a decline of A 1 R mRNA levels 6 h after pristane injection that was parallel to lymphocyte reduction. Following pristane,

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Cell-free DNA in plasma as an essential immune system regulator

Cited by 28 publications

References 61 publications

Validating quantitative PCR assays for cfDNA detection without DNA extraction in exercising SLE patients

Validating quantitative PCR assays for cfDNA detection without DNA extraction in exercising SLE patients

Circulating extracellular DNA is in association with continuous metabolic syndrome score in healthy adolescents

A1 and A2A adenosine receptors play a protective role to reduce prevalence of autoimmunity following tissue damage

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