Abstract
Background: Recently cell-free plasma DNA has been described as a marker of apoptosis during hemodialysis (HD), but little is known about how different dialysis membranes may contribute to this process or whether pre-HD levels are restored afterwards. Here we evaluate the influence of the dialysis membrane on cell-free plasma DNA levels and investigate the clearance of plasma circulating DNA after HD.
Methods: Cell-free plasma DNA was measured using a real-time quantitative PCR for the β-globin gene. Reference values for plasma DNA were established in a group of 100 healthy voluntary blood donors. Pre- and post-HD levels were also measured in 30 patients with end-stage renal disease on regular HD (52 sessions; 104 samples). The sessions lasted for 2.5–5h. Different dialysis membranes were compared: high-flux (n=37) vs. low-flux (n=15) and polysulfone (n=42) vs. modified cellulose (n=10). To determine the time at which pre-HD levels are restored, DNA was quantified in serial plasma samples obtained from 10 of these 30 patients, just before and immediately after HD, as well as at 30, 60 and 120min after HD.
Results: Reference plasma DNA values for healthy blood donors ranged from 112 to 2452gEq/mL (median 740gEq/mL). Cell-free plasma DNA levels significantly increased during HD (Wilcoxon test for paired samples, p<0.0001), with increases of more than four-fold observed in 75% of the patients after HD. There was no significant linear association between the length of the HD session (between 2.5 and 5h) and the increase in cell-free plasma DNA concentration (Pearson correlation). No significant differences were observed between different types of membranes (Mann-Whitney U-test). Plasma DNA returned to pre-HD levels by 30min after HD, regardless of the starting concentration.
Conclusions: Plasma DNA levels significantly increase after a conventional 2.5–5-h HD session. Therefore, HD patients require special consideration for correct interpretation of plasma DNA concentrations. This parameter can be considered a reliable diagnostic tool for certain pathologies when measured at least 30min after a HD session without further complications. The different dialysis membranes used in this study had no influence on cell-free plasma DNA concentrations, so the level of circulating DNA is not an appropriate marker of dialysis membrane biocompatibility.
Clin Chem Lab Med 2006;44:1410–5.
References
1. Mandel P, Metais P. Les acides nucleiques du plasma sanguine chez l'homme. CR Acad Sci Paris 1948; 142:241–3.Search in Google Scholar
2. Tong YK, Lo YM. Diagnostic developments involving cell-free (circulating) nucleic acids. Clin Chim Acta 2006; 363:187–96.10.1016/j.cccn.2005.05.048Search in Google Scholar
3. Taback B, Hoon DS. Circulating nucleic acids in plasma and serum: past, present and future. Curr Opin Mol Ther 2004; 6:273–8.Search in Google Scholar
4. Wu TL, Zhang D, Chia JH, Tsao KH, Sun CF, Wu JT. Cell-free DNA: measurement in various carcinomas and establishment of normal reference range. Clin Chim Acta 2002; 321:77–87.10.1016/S0009-8981(02)00091-8Search in Google Scholar
5. Lo YM, Tein MS, Lau TK, Haines CJ, Leeung TM, Poon PM, et al. Quantitative analysis of fetal DNA in maternal plasma and serum: implications for noninvasive prenatal diagnosis. Am J Hum Genet 1998; 62:768–75.10.1086/301800Search in Google Scholar
6. Rainer TH, Wong LK, Lam W, Yuen E, Lam NY, Metreweli C, et al. Prognostic use of circulating plasma nucleic acid concentrations in patients with acute stroke. Clin Chem 2003; 49:562–9.10.1373/49.4.562Search in Google Scholar
7. Lo YM, Rainer TH, Chan LY, Hjelm NM, Cocks RA. Plasma DNA as a prognostic marker in trauma patients. Clin Chem 2000; 46:319–23.10.1093/clinchem/46.3.319Search in Google Scholar
8. Lo YM, Tein MS, Pang CC, Yeung CK, Tong KL, Hjelm NM. Presence of donor-specific DNA in plasma of kidney and liver-transplant recipients. Lancet 1998; 351:1329–30.10.1016/S0140-6736(05)79055-3Search in Google Scholar
9. Jahr S, Hentze H, Englisch S, Hardt D, Fackelmayer FO, Hesch RD, et al. DNA fragments in the blood plasma of cancer patients: quantitation and evidence for their origin from apoptotic and necrotic cells. Cancer Res 2001; 61:1659–65.Search in Google Scholar
10. Stroun M, Lyautey J, Lederrey C, Olson-Sand A, Anker P. About the possible origin and mechanism of circulating DNA apoptosis and active DNA release. Clin Chim Acta 2001; 313:139–42.10.1016/S0009-8981(01)00665-9Search in Google Scholar
11. Gadi VK, Nelson JL, Boespflug ND, Guthrie KA, Kuhr C. Soluble donor DNA concentrations in recipient serum correlate with pancreas-kidney rejection. Clin Chem 2006; 52:379–82.10.1373/clinchem.2005.058974Search in Google Scholar PubMed
12. Zhong XY, Hahn D, Troeger C, Klemm A, Stein G, Thomson P, et al. Cell-free DNA in urine: a marker for kidney graft rejection, but not for prenatal diagnosis? Ann NY Acad Sci 2001; 945:250–7.10.1111/j.1749-6632.2001.tb03893.xSearch in Google Scholar
13. Atamaniuk J, Ruzicka K, Stuhlmeier KM, Karimi A, Eigner M, Mueller MM. Cell-free plasma DNA: a marker for apoptosis during hemodialysis. Clin Chem 2006; 52:523–6.10.1373/clinchem.2005.058883Search in Google Scholar PubMed
14. Steinman CR, Ackad A. Appearance of circulating DNA during hemodialysis. Am J Med 1977; 62:693–7.10.1016/0002-9343(77)90872-5Search in Google Scholar
15. Fournie GJ, Lule J, Dueymes JM, Laval F, Delobbe I, Vernier I, et al. Plasma DNA in patients undergoing hemodialysis or hemofiltration: cytolysis in artificial kidney is responsible for the release of DNA in circulation. Am J Nephrol 1989; 9:384–91.10.1159/000168000Search in Google Scholar PubMed
16. Ministerio de Sanidad y Consumo-Instituto de Salud Carlos III. Agencia de Evaluación de Tecnologías Sanitarias (AETS). Evaluación de los diferentes tipos de membranas de hemodiálisis. June 1996.Search in Google Scholar
17. Kaplow LS, Goffinet JA. Profound neutropenia during the early phase of hemodialysis. J Am Med Assoc 1968; 203:1135–7.10.1001/jama.1968.03140130047014Search in Google Scholar
18. Craddock PR, Fehr J, Dalmasso AP, Brighan KL, Jacob HS. Hemodialysis leukopenia. Pulmonary vascular leukostasis resulting from complement activation by dialyzer cellophane membranes. J Clin Invest 1977; 59:879–88.10.1172/JCI108710Search in Google Scholar PubMed PubMed Central
19. Panichi V, Migliori M, De Pietro S, Taccola D, Andreini B, Metelli MR, et al. The link of biocompatibility to cytokine production. Kidney Int 2000; 76 (Suppl):S96–103.10.1046/j.1523-1755.2000.07612.xSearch in Google Scholar PubMed
20. Pertosa G, Grandaliano G, Gesualdo L, Schena FP. Clinical relevance of cytokine production in hemodialysis. Kidney Int 2000; 76 (Suppl):S104–11.10.1046/j.1523-1755.2000.07613.xSearch in Google Scholar PubMed
21. Ghysen J, De Plaen JF, van Ypersele de Strihou C. The effect of membrane characteristics on tumour necrosis factor kinetics during haemodialysis. Nephrol Dial Transplant 1990; 5:270–4.10.1093/ndt/5.4.270Search in Google Scholar PubMed
22. Schindler R, Ertl T, Beck W, Lepenies J, Boenisch O, Oppermann M, et al. Reduced cytokine induction and removal of complement products with synthetic hemodialysis membranes. Blood Purif 2006; 24:203–11.10.1159/000090520Search in Google Scholar PubMed
23. Holdenrieder S, Stieber P, Bodenmuller H, Busch M, Von Pawel J, Schalhorn A, et al. Circulating nucleosomes in serum. Ann NY Acad Sci 2001; 945:93–102.10.1111/j.1749-6632.2001.tb03869.xSearch in Google Scholar PubMed
24. Gauthier VJ, Tyler LN, Mannik M. Blood clearance kinetics and liver uptake of mononucleosomes in mice. J Immunol 1996; 156:1151–6.10.4049/jimmunol.156.3.1151Search in Google Scholar
25. Kobayashi N, Kuramoto T, Yamaoka K, Hashida M, Takakura Y. Hepatic uptake and gene expression mechanisms following intravenous administration of plasmid DNA by conventional and hydrodynamics-based procedures. J Pharmacol Exp Ther 2001; 297:853–60.Search in Google Scholar
26. Botezatu I, Serdyuk O, Potapova G, Shelepov V, Alechina R, Molyaka Y, et al. Genetic analysis of DNA excreted in urine: a new approach for detecting specific genomic DNA sequences from cells dying in an organism. Clin Chem 2000; 46:1078–84.10.1093/clinchem/46.8.1078Search in Google Scholar
27. Lo YMD, Zhang J, Leung TN, Lau TK, Chang AM, Hjelm NM. Rapid clearance of fetal DNA from maternal plasma. Am J Hum Genet 1999; 64:218–24.10.1086/302205Search in Google Scholar PubMed PubMed Central
28. Lau TW, Leung TN, Chan LY, Lau TK, Chan KC, Tam WH, et al. DNA clearance from maternal plasma is impaired in preeclampsia. Clin Chem 2002; 48:2141–6.10.1093/clinchem/48.12.2141Search in Google Scholar
29. Burlingame RW, Volzer MA, Harris J, Du Clos TW. The effect of acute phase proteins on clearance of chromatin from the circulation of normal mice. J Immunol 1996; 156:4783–8.10.4049/jimmunol.156.12.4783Search in Google Scholar
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