Two pathogenetically distinct disease manifestations are distinguished in a murine model of primary rabies virus infection with the Evelyn-Rokitnicky-Abelseth strain, rabies virus neuritic paralysis (RVNP) and fatal encephalopathogenic rabies. RVNP develops with high incidence in immunocompetent mice after intraplantar infection as a flaccid paralysis restricted to the infected limb. The histopathologic correlate of this monoplegia is a degeneration of the myelinated motor neurons of the peripheral nerve involved. While, in this model, fatal encephalopathogenic rabies develops only after depletion of the CD4 subset of T lymphocytes and without contribution of the CD8 subset, RVNP is identified as an immunopathological process in which both the CD4 and CD8 subsets of T lymphocytes are critically implicated.
Previously, we have determined that human annexin V (hAV), a Ca2+-dependent phospholipid-binding protein, and not rat AV, binds specifically to small hepatitis B surface antigen (SHBsAg), and that transfection of a rat hepatoma cell line with a construct containing the hAV gene led to hAV expression and conferred susceptibility to hepatitis B virus (HBV) infection. In this work, we have examined the effect of administration of hAV on in vitro binding of SHBsAg to human and to rat hepatocytes and on in vitro HBV infection. The results showed that hAV could bind to human as well as to rat hepatocytes. Because of this property, excess hAV was unable to prevent HBV infection in primary cultures of human hepatocytes. On the other hand, it enabled rat hepatocytes to specifically bind SHBsAg and conferred susceptibility to HBV infection. After infection of primary cultures of rat hepatocytes in the presence of hAV, HBV mRNA, covalently closed circular (ccc) DNA, replicative intermediates, hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg) and secreted HBV DNA were detected. After infection in the absence of hAV, no markers of HBV replication were detected. Hence, from the present study we conclude that hAV is involved in facilitating HBV entry, leading to successful HBV infection in primary cultures of rat hepatocytes, while it is not effective in preventing HBV infection in primary cultures of human hepatocytes.
Summary
Hybridomas were prepared by fusion of spleen cells from BALB/c mice immunized with dog rabies isolates from Nigeria with P3times63Ag8 myeloma cells. More than 69 hybridomas secreted antinucleocapsid (antiNC) antibodies when tested with homologous viruses by indirect immunofluorescence. One hybridoma (Z144‐88) was found which secreted antiNC antibody that reacted negatively with fox rabies isolates from the Federal Republic of Germany, Switzerland and France and with rabies‐related viruses and European bat isolates. It reacted positively with other strains/isolates of rabies virus. It is possible to use this antiNC monoclonal antibody (mab) for the investigation of fox rabies outbreaks in Europe.
Zusammenfassung
Hybridoma‐Zellen wurden hergestellt durch Fusion von P3times63Ag8‐Myelomazellen und Milzzellen aus BALB/c‐Mäusen, die mit Hundetollwutisolaten aus Nigeria immunisiert wurden. Mehr als 69 Hybridoma‐Zelltypen produzierten Antinukleokapsid monoklonale Antikörper, wenn sie mit homologen Viren im indirekten Fluoreszenztest geprüft wurden. Ein Hybridoma‐Zelltyp (Z144‐88) produzierte Antinukleokapsid monoklonale Antikörper, die negativ mit Fuchstollwut‐Isolaten aus der Bundesrepublik Deutschland, der Schweiz und Frankreich und mit verschiedenen europäischen Fledermaustollwut‐Isolaten reagierten. Sie reagierten positiv mit anderen Stämmen / Isolaten der Tollwutvirusgruppe. Diese Antinukleokapsid monoklonale Antikörper können zur Erkennung von Fuchstollwut‐Ausbrüchen in Europa herangezogen werden.
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