Peterlana, et al: Parvovirus B19 DNA in RA 1907 From the Department of Clinical and Experimental Medicine, Section of Internal Medicine, and the Department of Anesthesiology and Surgery, Section of Orthopedics, University of Verona; the Department of Experimental Medicine, University of Genova; and G. Gaslini Institute, Genova, Italy. Supported by grants from the MURST 60%, University of Verona and Genova, and from Cariverona Foundation. D. Peterlana, MD; R. Beri, BS; S. Simeoni, MD; L. Borgato, PhD; L. Scilanga, MD; S. Cerù, MD; R. Corrocher, MD, Full Professor of Internal Medicine; C. Lunardi, MD, Associate Professor, Department of Clinical and Experimental Medicine, Section of Internal Medicine, University of Verona; A. Puccetti, MD, Associate Professor, Department of Experimental Medicine, University of Genova; M. Ricci, MD, Department of Anesthesiology and Surgery, Section of Orthopedics, University of Verona. Address reprint requests to Dr. C. Lunardi, Department of Clinical and Experimental Medicine, Section of Internal Medicine Policlinico GB Rossi-P. le LA Scuro, 37134, Verona, Italy. E-mail: claudio.lunardi@univr.it Submitted October 23, 2002; revision accepted February 3, 2003. Rheumatoid arthritis (RA) is an autoimmune disease of unknown etiology, characterized by chronic inflammation of the synovial membrane (SM) eventually leading to joint destruction. Both genetic and environmental factors are believed to be involved in the pathogenesis of RA, and among the latter several viral agents have been proposed as possible triggers of the disease 1,2 . On the basis of paleopathological analysis 3 , several investigators have focused their attention on parvovirus B19 (B19) which can be responsible for a symmetric arthritis in adults that can last for a few weeks 4 ; on some occasions the infection can persist, leading to a chronic arthritis resem- bling RA 5 . The virus can induce an autoimmune response and some of us have shown that anti-virus antibodies are able to cross-react with autoantigens 6 . However the search for B19 DNA in synovial tissue with a very sensitive tech- nique such as the polymerase chain reaction (PCR) has yielded conflicting results 7-11 . The B19 genome encodes for 2 structural proteins VP1 and VP2 and for the non structural protein NS1, which seems to be essential for viral packaging 12 . Recently, some investigators have reported that NS1 is cytotoxic to hemopoietic cells 13 and is able to induce over- expression of the proinflammatory cytokine interleukin (IL)-6 gene in vitro 14 . Moreover, NS1 has been associated with the persistence of the virus 7 , and antibodies directed against NS1 may therefore be used as a marker of chronic infection 15,16 . Most of the studies published so far have evaluated the persistence of B19 by amplifying the VP genes using nested PCR, or the NS1 gene using single round-PCR. In consider- ation of the possible importance of NS1 gene in chronic The Presence of Parvovirus B19 VP and NS1 Genes in the Synovium Is Not Correlated with Rheumatoid Arthritis DIMITRI PETERLANA, ANTONIO PUCCETTI, RUGGERO BERI, MATTEO RICCI, SARA SIMEONI, LORENA BORGATO, LUCIA SCILANGA, SILVIA CERÙ,ROBERTO CORROCHER, and CLAUDIO LUNARDI ABSTRACT. Objective. To amplify both NS1 and VP genes of Parvovirus B19 DNA in synovial membrane (SM) and serum obtained from patients with rheumatoid arthritis (RA) and to analyze whether the pres- ence of viral DNA is correlated with synovitis. Methods. DNA obtained from 30 SM and 24 serum samples from RA patients was analyzed using single round-polymerase chain reaction (PCR) and nested PCR for both VP and NS1 genes of parvovirus B19. Twenty-four SM and serum samples from sex and age matched subjects with osteoarthritis (OA) or joint trauma served as controls. Results. The first round PCR was negative for NS1 in RA samples. After nested PCR, NS1 was detected in the SM of 6/30 patients and of 10/24 controls and in the serum of 4/24 patients and controls. Nested PCR for the VP gene detected viral DNA in the SM of 7/30 patients with RA and of 7/24 of the controls and in the serum of 5/24 patients and of 2/24 controls. Altogether parvovirus DNA was found in the SM of 11/30 (36.6%) patients and of 12/24 (50%) controls and in the serum of 8/24 (33.3%) patients with RA and of 5/24 (20.8%) controls. Conclusion. Our results suggest that the amplification by nested PCR of both NS1 and VP genes is necessary to define the presence of viral DNA in tissue samples and confirm that the presence of parvovirus B19 DNA is similar in RA and control SM, suggesting that simple detection of viral DNA is not sufficient to confirm a link between the virus and RA. (J Rheumatol 2003;30:1907–10) Key Indexing Terms: PARVOVIRUS B19 NS1 AND VP GENES RHEUMATOID SYNOVITIS Personal, non-commercial use only. The Journal of Rheumatology Copyright © 2003. All rights reserved. www.jrheum.org Downloaded on December 8, 2021 from