title: Parvoviral infection of endothelial cells and stromal fibroblasts: a possible pathogenetic role in scleroderma.
abstract: BACKGROUND
  Systemic sclerosis (SSc) is a connective tissue disease (CTD) which differs from other CTDs by progressive irreversible fibrosis in lung, kidney, skin, and heart. It has a worse prognosis compared to several other CTDs. The pathogenesis may reflect a humorally mediated microangiopathy in concert with the overproduction of collagen triggered by immune-mediated cytokine production. Having previously demonstrated parvovirus B19 (B19) DNA in bone marrow and skin biopsies of SSc patients in the absence of B19 viremia, we sought to further elucidate a role for B19 in the pathogenesis of SSc.
  
  
  METHODS
  Twelve patients who fulfilled American College of Rheumatology criteria for a diagnosis of SSc were encountered. Ten were serologically screened for B19 infection. Solution phase polymerase chain reaction (PCR) for B19 DNA was performed on skin tissue from six patients, and in all biopsies, reverse transcriptase in situ PCR (RT in situ PCR) for B19 and tumor necrosis factor (TNF)-alpha mRNA was performed. B19 viral protein (VP2) expression was sought by immunohistochemistry and correlated to PCR findings and to light microscopy of hematoxylin and eosin-stained sections. Frozen tissue was also available on five of the patients. Two control groups were assessed for B19 and TNF expression comprising one with irrelevant primers and the other representing 18 cases of inflammatory skin lesions where the etiology was known and unrelated to B19 infection. In addition, frozen and paraffin-embedded tissues procured from skin lesions unrelated to B19 infection were assessed for B19 genome. In all cases, pretreatment with RNase was also performed to verify that any positive signal was indeed RNA based.
  
  
  RESULTS
  Diffuse SSc was seen in seven patients, and limited disease in five. All patients had an antinuclear antibody--specifically, an antinucleolar, anticentromere, and/or anti-Scl 70 antibody. Eleven of the 12 had lung involvement, whereas eight patients had myocardial disease. Of 12 patients tested serologically, nine had B19-specific antibodies, which included immunoglobulin M (IgM)-specific antibodies in two cases. Solution phase PCR showed B19 DNA in the skin in three cases and in the bone marrow in three cases, including two in whom skin-based B19 DNA was observed. In all cases, RT in situ PCR demonstrated B19 and TNF-alpha mRNA in endothelia, fibroblasts, mast cells, and perivascular inflammatory cells. Immunohistochemistry to assess VP2 was either negative or equivocal. Immunofluorescent studies revealed prominent deposition of C5b-9 within the cutaneous vasculature from biopsies of all patients tested. The control samples were negative for B19 and TNF RNA and DNA.
  
  
  CONCLUSIONS
  Parasitism of endothelia and fibroblasts by B19 with resultant enhanced TNF-alpha expression may be of pathogenetic importance in SSc even in the absence of demonstrable viremia. The vascular deposition of C5b-9 suggests a role for humoral immunity possibly induced by a state of endothelial neoantigenicity evoked by virally mediated cell injury. Treatment strategies include anti-viral therapy, including in the context of intravenous gamma-globulin and anti-TNF therapy.
authors.affiliation: Department of Pathology, The Ohio State University, Columbus, OH, USA.
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Magro
authors.forename: Gerard
authors.initials: G
authors.lastname: Nuovo
authors.forename: Clodoveo
authors.initials: C
authors.lastname: Ferri
authors.forename: A Neil
authors.initials: AN
authors.lastname: Crowson
authors.forename: Dilia
authors.initials: D
authors.lastname: Giuggioli
authors.forename: Marco
authors.initials: M
authors.lastname: Sebastiani
mesh_terms: D000328:Adult; D000368:Aged; D036022:Capsid Proteins / Q000378:metabolism; D015938:Complement Membrane Attack Complex / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology / Q000821:virology; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D008297:Male; D008875:Middle Aged; D010322:Parvoviridae Infections / Q000150:complications* / Q000473:pathology; D016732:Parvovirus B19, Human / Q000235:genetics / Q000472:pathogenicity*; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045743:Scleroderma, Diffuse / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D045745:Scleroderma, Limited / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D017154:Stromal Cells / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D015938:Complement Membrane Attack Complex; D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; D014409:Tumor Necrosis Factor-alpha; C095548:capsid protein VP2, parvovirus B19