Lack of HHV-8 DNA Sequences in Sarcoid Tissues of French Patients

Abstract

Recently, Di Alberti and colleagues reported an association between human herpesvirus (HHV)-8 and sarcoidosis.1Di Alberti L Piattelli A Artese L et al.Human herpesvirus 8 variants in sarcoid tissues..Lancet. 1997; 350: 1655-1661Abstract Full Text Full Text PDF PubMed Scopus (157) Google Scholar HHV-8 DNA sequences were detected by a nested polymerase chain reaction (PCR) procedure, amplifying a segment of the open reading frame (ORF) 26, in 38 of 39 sarcoid tissues from 17 patients with sarcoidosis, and in 6 of 113 tissues from 96 patients without sarcoidosis. We looked for HHV-8 DNA sequences by nested PCR in biopsy samples of French patients with and without sarcoidosis. We evaluated frozen tissue samples with histological evidence of sarcoid granuloma obtained from 14 patients with systemic sarcoidosis kept at – 80°C until analysis, including 10 mediastinal lymph nodes and four deltoid muscle biopsy samples. Nonsarcoid tissues, including superficial lymph node frozen biopsy samples showing mild reactive changes from 17 non-HIV-infected patients without sarcoidosis, superficial lymph node paraffin-embedded samples showing Castleman's disease from 8 non-HIV-infected patients with multicentric Castleman's disease, and frozen Kaposi's sarcoma tissues from 3 HIV 1-infected patients were used as control subjects. Two nested PCR amplifying segments in ORF252Moore PS Kingsley LA Holmberg SD et al.Kaposi's sarcoma-associated herpesvirus infection prior onset of Kaposi's sarcoma..AIDS. 1996; 10: 175-180Crossref PubMed Scopus (299) Google Scholar and in ORF263Zong J-C Metroka C Reitz MS et al.Strain variability among Kaposi sarcoma-associated herpesvirus (human herpesvirus 8) genomes: evidence that a large cohort of United States AIDS patients may have been infected by a single common isolate..J Virol. 1997; 71: 2505-2511PubMed Google Scholar were performed in parallel in 1 μg DNA obtained by phenolchloroform extraction, after removal of paraffin by xylene when necessary. All samples were positive by PCR for human β-globin gene, assessing optimal DNA extraction and lack of PCR inhibitors. Both nested PCR were positive in tissues of three of three patients with Kaposi's sarcoma, and in six of eight patients with multicentric Castleman's disease. Two of 17 nonsarcoid lymph node tissues showed possible presence of HHV-8, the ORF25 nested PCR being positive. None of the 14 sarcoid tissues were found positive for HHV-8 ORF25 or ORF26 DNA sequences. Our findings are in keeping with previous studies showing that HHV-8 DNA sequences are constantly detected in patients with Kaposi's sarcoma,4Chang Y Cesarman E Pessin MS et al.Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma..Science. 1994; 266: 1865-1869Crossref PubMed Scopus (4982) Google Scholar frequently detected in patients with multicentric Castleman's disease,5Soulier J Grollet L Ocksenhendler E et al.Kaposi's sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman's disease..Blood. 1995; 86: 1276-1280PubMed Google Scholar and occasionally detected in reactive lymph nodes from non-HIV-infected patients.6Luppi M Barozzi P Maiorana A et al.Human herpesvirus-8 DNA sequences in human immunodeficiency virus-negative angioimmunoblastic lymphadenopathy and benign lymphadenopathy with giant germinal center hyperplasia and increased vascularity..Blood. 1996; 87: 3903-3909PubMed Google Scholar In contrast, failure to detect HHV-8 DNA sequences in histologically proven sarcoid tissues from 14 patients with systemic sarcoidosis does not support an association between HHV-8 and sarcoidosis.