Increased levels of circulating DNA in patients with systemic autoimmune diseases: A possible marker of disease activity in Sjögren’s syndrome

Abstract

High levels of serum and/or plasma circulating DNA (cDNA) have been described in patients with systemic autoimmune diseases (SADs). However, the role of this molecule has not been clarified. Our aim was to evaluate plasma cDNA levels in 48 systemic lupus erythematosus (SLE) and 44 primary Sjögren’s syndrome (SS) patients, as compared with healthy and rheumatoid arthritis (RA) subjects, and to analyse their correlation with disease activity, disease damage and clinical manifestations. Plasma DNA was extracted using Qiagen columns and quantified by real-time quantitative PCR. Disease activity and damage were evaluated in both diseases by analysis of clinical and laboratory findings. Our results showed that plasma cDNA levels were significantly higher in patients with SS (mean ± SE: 32.0 ± 7.3 ng/ml) and with SLE (35.0 ± 9.0 ng/ml) than in controls (5.1 ± 1.1 ng/ml) (p < 0.0001 for both). Disease activity index correlated with cDNA levels in SS (p = 0.02), but not in SLE, and SS subjects with active disease displayed significantly higher cDNA levels with respect to inactive patients (p < 0.05). No correlation was found between plasma cDNA levels and disease damage indexes in either SLE or SS. These results indicate that increased plasma cDNA levels can been demonstrated in SLE and in SS patients with respect to healthy subjects. Interestingly, although cDNA levels did not correlate with indexes of disease damage in these disorders, a significant correlation between cDNA concentrations and disease activity was observed in SS, but not in SLE, suggesting a possible role of cDNA as non-invasive marker of disease activity. The different results obtained in these SADs may be explained by distinct disease pathogenesis or the influence of immunosuppressive and corticosteroid therapy that, unlike in SS, is usually employed in SLE.

Keywords

disease activity Sjögren’s syndrome systemic lupus erythematosus

Get full access to this article

View all access options for this article.

References

Mandel

Metais

. Les acides nucleiques du plasma sanguine chez l’homme. CR Acad Sci Paris 1947; 142: 241–3.

Yoon

Park

Lee

Kim

Lee

. Comparison of circulating plasma DNA levels between lung cancer patients and healthy controls. J Mol Diagn 2009; 11: 182–5.

Ludovini

Pistola

Gregorc

. Plasma DNA, micro satellite alterations, and p53 tumor mutation are associated with disease-free survival in radically resected non-small cell lung cancer patients. J Thorac Oncol 2008; 3: 365–373.

Zhang

Shao

Ying

. Value of quantitative analysis of circulating cell free DNA as a screening tool for lung cancer: a meta-analysis. Lung Cancer 2010; 69: 225–231.

Tsang

. Circulating nucleic acids in plasma/serum. Pathology 2007; 39: 197–207.

Uzuelli

Dias-Junior

Izidoro-Toledo

Gerlach

Tanus-Santos

. Circulating cell-free DNA levels in plasma increase with severity in experimental acute pulmonary thromboembolism. Clin Chim Acta 2009; 409: 112–116.

Suzuki

Kamataki

Yamaki

Homma

. Characterization of circulating DNA in healthy human plasma. Clin Chim Acta 2008; 387: 55–58.

Tan

Schur

Carr

Unkel

. Deoxyribonucleic acid [DNA] and antibodies to DNA in the serum of patients with systemic lupus erythematosus. J Clin Invest 1966; 45: 1732–1740.

Raptis

Menard

. Quantitation and characterization of plasma DNA in normals and patients with systemic lupus erythematosus. J Clin Invest 1980; 66: 1391–1399.

10.

Klemp

Meyers

Harley

. Measurement of plasma DNA by a physicochemical method: relevance in SLE. Ann Rheum Dis 1981; 40: 593–599.

11.

Leon

Ehrlich

Shapiro

Labbate

. Free DNA in the serum of rheumatoid arthritis patients. J Rheumatol 1977; 4: 139–142.

12.

McCoubrey-Hoyer

Okarma

Holman

. Partial purification and characterization of plasma DNA and its relation to disease activity in systemic lupus erythematosus. Am J Med 1984; 77: 23–34.

13.

Chen

Meister

Urbonaviciute

. Sensitive detection of plasma/serum DNA in patients with systemic lupus erythematosus. Autoimmunity 2007; 40: 307–310.

14.

Mosca

Giuliano

Cuomo

. Cell-free DNA in the plasma of patients with systemic sclerosis. Clin Rheumatol 2009; 28: 1437–1440.

15.

Pisetsky

. DNA as a marker of cell death in systemic lupus erythematosus. Rheum Dis Clin N Am 2004; 30: 575–587.

16.

van der Vaart

Pretorius

. Circulating DNA. Its origin and fluctuation. Ann N Y Acad Sci 2008; 1137: 18–26.

17.

Pisetsky

Lipsky

. Microparticles as autoadjuvants in the pathogenesis of SLE. Nat Rev Rheumatol 2010; 6: 368–372.

18.

Steinman

. Circulating DNA in systemic lupus erythematosus. Association with central nervous system involvement and systemic vasculitis. Am J Med 1979; 67: 429–435.

19.

Vitali

Bombardieri

Jonsson

. , for European Study Group on Classification Criteria for Sjögren’s Syndrome. Classification criteria for Sjögren’s syndrome: a revised version of the European criteria proposed by the American–European Consensus Group. Ann Rheum Dis 2002; 61: 554–558.

20.

Tan

Cohen

Fries

. The 1982 revised criteria fort he classification of systemic lupus erythematosus. Arthritis Rheum 1982; 25: 1271–1277.

21.

Arnett

Edworthy

Bloch

. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 1988; 31: 315–324.

22.

Vitali

Palombi

Baldini

. Sjögren’s Syndrome Disease Damage Index and disease activity index: scoring systems for the assessment of disease damage and disease activity in Sjögren’s syndrome, derived from an analysis of a cohort of Italian patients. Arthritis Rheum 2007; 56: 2223–2231.

23.

Bombardier

Gladman

Urowitz

Caron

Chang

DH,

. and the Committee on Prognosis Studies in SLE Derivation of the SLEDAI: a disease activity index for lupus patients. Arthritis Rheum 1992; 35: 630–640.

24.

Stoll

Seifert

Isenberg

. SLICC/ACR Damage Index is valid, and renal and pulmonary organ scores are predictors of severe outcome in patients with systemic lupus erythematosus. Br J Rheumatol 1996; 35: 248–254.

25.

Gladman

Ibanez

Urowitz

. Systemic lupus erythematosus disease activity index 2000. J Rheumatol 2002; 29: 288–291.

26.

Heid

Stevens

Livak

Williams

. Real time quantitative PCR. Genome Res 1996; 6: 986–994.

27.

Holland

Abramson

Watson

Gelfand

. Detection of specific polymerase chain reaction product by utilizing the 5′-3′ exonuclease activity of Thermus acquaticus DNA polymerase. Proc Natl Acad Sci USA 1991; 88: 7276–7280.

28.

Steinman

. Circulating DNA in systemic lupus erythematosus. Isolation and characterization. J Clin Invest 1984; 73: 832–841.

29.

Lee

Montalvo

Chrebtow

Busch

. Quantitation of genomic DANN in plasma and serum samples: higher concentration of genomic DANN found in serum than in plasma. Transfusion 2001; 41: 276–282.

30.

Zeerleder

. The struggle to detect circulating DNA. Crit Care 2006; 10: 142–144.

31.

Rumore

Steinman

. Endogenous circulating DNA in systemic lupus erythematosus. J Clin Invest 1990; 86: 69–74.

32.

Chen

Fineschi

Morozzi

Marcolongo

Chen

Galeazzi

. Modifications of DANN patterns in plasma and nucleated blood cells from systemic sclerosis patients. Clin Exp Rheumatol 2001; 19: 492–494.

33.

Galeazzi

Morozzi

Piccini

. Dosage and characterization of circulating DNA: present usage and possible applications in systemic autoimmune disorders. Autoimmunity Rev 2003; 2: 50–55.

34.

Pisetsky

Jiang

. The generation of extracellular DNA in SLE: the role of death and sex. Scand J Immunol 2006; 64: 200–204.

35.

Pisetsky

. The immune response to cell death in SLE. Autoimmunity Rev 2004; 3: 500–504.

36.

Jimenez

Aiba-Masago

Al Hashimi

. Activated caspase 3 and cleaved poly [ADP-ribose] polymerase in salivary epithelium suggest a pathogenetic mechanism for Sjögren’s syndrome. Rheumatology 2002; 41: 338–342.

37.

Sisto

Lisi

Lofrumento

D’Amore

Scagliusi

Mitolo

. Autoantibodies from Sjögren’s syndrome trigger apoptosis in salivary gland cell line. Ann N Y Acad Sci 2007; 1108: 418–425.

38.

Hughes

Cohen

Lightfoot

Meltzer

Christian

. The release of DNA into serum and synovial fluid. Arthritis Rheum 1971; 14: 259–266.

39.

Bryzgunova

Skvortsova

Kolesnikova

. Isolation and comparative study of cell-free nucleic acids from human urine. Ann N Y Acad Sci 2006; 1075: 334–340.

40.

Korabecna

Opatrna

Wirth

. Cell-free plasma DANN during peritoneal dialysis and hemodialysis and in patients with chronic kidney disease. Ann N Y Acad Sci 2008; 1137: 296–301.

41.

Porciello

Scarpato

Ferri

. Spontaneous chromosome damage [micronuclei] in systemic sclerosis and Raynaud’s phenomenon. J Rheumatol 2003; 30: 1244–1247.