RESUMO
A retroviral element (MSRV) defining a family of genetically inherited endogenous retroviruses (HERV-W) has recently been characterized in cell cultures from patients with multiple sclerosis (MS). To address the possible relationship with MS, direct detection of circulating virion RNA was proposed but revealed technically difficult to perform in standardized conditions, in the face of multiple endogenous HERV-W copies. A parallel approach has evaluated MSRV potential pathogenicity in relation to characteristic features of multiple sclerosis, in particular, T-lymphocyte-mediated immunopathology. We report here that MSRV particles induce T-lymphocyte response with a bias in the Vbeta16 chain usage in surface receptor, whatever the HLA DR of the donor. A recombinant MSRV envelope-but not core-protein reproduced similar nonconventional activation. Molecular analysis of Vbeta CDR3 showed that Vbeta16 expansions are polyclonal. Our results thus provide evidence that MSRV envelope protein can trigger an abnormal immune response with similar characteristics to that of superantigens.
Assuntos
Retrovirus Endógenos/imunologia , Ativação Linfocitária/imunologia , Esclerose Múltipla/virologia , Infecções por Retroviridae/imunologia , Linfócitos T/imunologia , Proteínas do Envelope Viral/imunologia , Antígenos Virais/imunologia , Células Cultivadas , Citocinas/metabolismo , Retrovirus Endógenos/genética , Humanos , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/virologia , Esclerose Múltipla/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/metabolismo , Proteínas Recombinantes/imunologia , Infecções por Retroviridae/virologia , Linfócitos T/metabolismo , Células Tumorais Cultivadas , Vírion/imunologiaRESUMO
OBJECTIVES: To determine whether the recently identified multiple sclerosis-associated retrovirus, MSRV, is detectable in the serum and synovial fluid of patients with rheumatoid arthritis (RA). METHODS: A reverse transcription-polymerase chain reaction (RT-PCR) assay was used to seek evidence of particle-associated MSRV/HERV-W RNA in the plasma and synovial fluid of patients with RA and controls. Stringent precautions were taken to avoid detection of contaminating human genomic DNA and cellular RNA sequences. RESULTS: Thirty-seven plasma samples were tested (20 from RA patients and 17 from controls) but none had detectable MSRV/HERV-W RNA. Synovial fluid samples were available from nine patients with RA and 10 controls. Particle-associated MSRV/HERV-W RNA was reproducibly detected in two of nine synovial fluid samples from RA patients and in one control sample. The identity of RT-PCR products was confirmed by sequencing. CONCLUSION: MSRV/HERV-W RNA sequences are detectable in the synovial fluid of a small proportion of RA patients, but this phenomenon may not be specific to RA.
Assuntos
Artrite Reumatoide/virologia , Esclerose Múltipla/virologia , RNA/análise , Retroviridae/genética , Líquido Sinovial/química , HumanosRESUMO
New sequences have been obtained by successive overlapping RT-PCR extensions from the pol region of a retroviral RNA (multiple sclerosis-associated retroviral element, MSRV) amplified in retrovirus-like particles from patients with multiple sclerosis. gag and pol sequences are related to type C oncoviruses, whereas the env sequence is closer to type D. A tryptophan-like (W) tRNA primer-binding site was identified downstream of the RU5 region in the 5'LTR, and the U3R region cloned in the 3'LTR exhibited potent promoter activity. MSRV clones define a novel family of endogenous elements, HERV-W. From our data, HERV-W RNAs are copackaged in extracellular particles which might be produced by replication-competent or transcomplemented HERV-W copies or by an exogenous member of the HERV-W family.
Assuntos
Retrovirus Endógenos/genética , Esclerose Múltipla/virologia , Sequência de Aminoácidos , Sequência de Bases , Sítios de Ligação , Linhagem Celular , Clonagem Molecular , Códon de Terminação , Produtos do Gene env/metabolismo , Humanos , Integrases/metabolismo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Regiões Promotoras GenéticasAssuntos
Produtos do Gene gag/genética , Variação Genética , Antígenos HIV/genética , Proteína gp120 do Envelope de HIV/genética , Infecções por HIV/virologia , Transcriptase Reversa do HIV/genética , HIV-1/genética , Fragmentos de Peptídeos/genética , Proteínas Virais , Sequência de Aminoácidos , Seguimentos , Infecções por HIV/transmissão , Humanos , Dados de Sequência Molecular , Produtos do Gene gag do Vírus da Imunodeficiência HumanaRESUMO
Among 47 blood donors tested positive with HCV EIA 2.0 Abbott, 27 (57.4%) also reacted with four ¿third-generation' EIAs. The presence of anti-HCV antibodies was confirmed with 3 different immunoblot assays in 16 of 27 sera (34.0%) while 10 samples (21.3%) had indeterminate profile with antibodies usually directed against structural core antigen. Anti-HCV core IgM response was found in 12 of 47 sera (25.5%) and HCV viremia detected by the polymerase chain reaction (PCR) procedure was observed in 15 samples (31.9%). A comparative study of the different markers confirmed a good correlation between a strong antibody response in EIAs and immunoblot assays and the presence of HCV RNA in the serum; only 2 immunoblot indeterminate samples were PCR positive. An association was observed between IgM antibodies against "core' epitopes and HCV RNA carriage: all IgM-positive sera were found positive by PCR. However, the direct detection of viral genome remains the best method for identifying HCV carriers in the blood donor population.