Abstract

Circulating cell-free DNA (cfDNA) may be involved in immune response regulation. We studied the variations in abundance of telomeric sequences in plasma and serum in young healthy volunteers and the ability of cfDNA contained in these samples to co-activate the TNF-α m RNA expression in monocytes. We performed qPCR to determine relative telomere length (T/S ratios) in plasma, serum and whole blood of 36 volunteers. Using paired samples of plasma and serum and DNase treatment, we analysed the contribution of cfDNA to the co-activation of TNF-α mRNA expression in THP1 monocytic cell line. We found significant differences between paired plasma and serum samples in relative T/S ratios (median 1.38 ± 1.1 vs. 0.86 ± 0.25, respectively) and in total amounts of cfDNA and in estimated total amounts of telomeres which were significantly higher in serum than in plasma. TNF-α mRNA expression in THP1 cells increased significantly after DNase treatment of all samples used for stimulation. The highest TNF-α mRNA expressions were observed after stimulation with DNase treated serum samples. Our results suggest that the different content of telomeric sequences in plasma and serum may contribute to the tuning of immune response. Further studies of this interesting phenomenon are needed.

Highlights

  • Telomeres are known to prevent deleterious shortening of linear DNA of eukaryotic chromosomes during DNA replication

  • It is known that serum samples contain more Cell-free DNA (cfDNA) than the paired plasma samples[17], examination of serum may be used to enhance the detection of cancer specific mutations in cfDNA18

  • We examined the effects of selected paired samples of plasma and serum on the stimulation of TNF-α (Tumor Necrosis Factor-α) mRNA expression in monocytic cell line THP1 before and after DNase treatment of these biological fluids

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Summary

Introduction

Telomeres are known to prevent deleterious shortening of linear DNA of eukaryotic chromosomes during DNA replication. The length of telomere sequences is studied under wide clinical conditions with the goal to find markers distinguishing different health statuses of patients and allowing prognosis of their clinical outcomes[4, 5]. The elevated levels of cfDNA or the detection of specific gene sequences in its pool in circulation are reported in numerous clinical conditions associated with inflammation, tissue damage and cancer[10,11,12,13]. The phenomenon of NETosis (Neutrophil Extracellular Trap) as quite recently described type of cell death[19] could potentially provide an explanation of the different concentrations of cfDNA in paired plasma and serum samples as well as at least in some clinical situations associated with inflammation in which cfDNA levels in circulation are increased[20]. Fuchs et al.[24] found that NETs are liberated during storage of non-leukoreduced red blood cells (RBC)

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