- Email: [email protected]
HBV DNA in peripheral mononuclear blood cells: Relationship with viral replication
113
SEROLOGIC PROFILE OF HEPATITIS D VIRUS (HDV) RELATED FULMINANT HEPATITIS. A.Mas, M. Buti, J. Costa, J.M.Sanchez-Tapias, R. Esteban, J.Guardia and J.Rod~s. Liver Unit, Hospital Cllnico and Dept. of Internal Medicine, Hospital Valle de Hebron, Barcelona, Spain.
To assess the diagnostlc value of HDV serum markers in fulminant hepatitis D a longitudinal study of the serologic p r o f i l e of 27 patients who were referred for HBsAg or IgM anti-HBc positive fulminant hepatic f a i l u r e has been performed. SerumHDAg, t o t a l a n t i HD, IgM anti-HD and RNA-VHD were examined every two to three days during the phase of severe l i v e r f a i l u r e and weekly thereafter in survivors. A single HDV immunological marker or a combination of markers was detected in the f i r s t serum sample from 18 patients. HBsAg was present in a l l and 14 of them also had IgM a n t i HBc. HDV markers were never detected in the nine remalning patients. HDAg alone was detected in six cases, HDAg and anti-HD in five ( t o t a l anti-HD in one and both t o t a l and IgM anti-HD in four) and a n t i - delta alone in seven ( t o t a l anti-HD in two and both t o t a l and IgM anti-HD in f i v e ) . Thus, isolated IgM anti-HD was never observed. HDV-RNA was detected only in three patients with concomitant HD antigenemia. All patients with HDAg in serum were admitted less than ten days after the beginning of symptoms. Among patients with HDAg alone, seroconversion to both t o t a l and IgM anti-HD occurred within a few days in three cases whlle isolated HDAg remained in three further patients who died shortly. In patients i n i t i a l l y showing both HDAg and anti-HD, the antigen became undetectable a few days l a t e r . Changes of the i n i t i a l serological pattern were not observed in patients with isolated anti-HD in the i n i t i a l sample. The precedent observations indicate that testing for both HDAg and t o t a l anti-HD is mandatory for the diagnosis of fulminant hepatitis D while IgM anti-HD and HDV-RNA have l l t t l e diagnostic value in this sltuation.
114
HBV DNA IN PERIPHERAL MONONUCLEARBLOOD CELLS: RELATIONSHIP WITH VIRAL REPLICATION M.Melegari,C.Pasquinelli,M.Solieri,F.Manenti,E.Villa Chair of Gastroenterology, University of Modena, Italy.
Hepatitis B virus (HBV) infection of peripheral mononuclear blood cells (PMBCS) has been recently reported as a Feature occurring in all stages of liver disease. Moreover, in the related woodchuck hepatitis virus model specific transcripts have been shown in FMBCs and in the spleen of chronically infected animals, suggesting that a complete replicative cycle can take place in htese cells. We therefore investigated 30 patients with acute type B hepatitis (6/30 HBV DNA+, 14/30 HBeAg+) and a smaller group of 5 patients with chronic active hepatitis HBeAg+, HBV DNA+,followed up at monthly intervals. Southern and Northern blot analysis of PMBCs were performed utilizing a Full lenght HBV probe. No HBV specific sequences in PMBCs were displayed in the acute hepatitis group either in Southern or Northern blot, although the sensitivity of our method enabled us to detect as little as 1 pg of cloned HBV insert. On the contrary,in 5/5 chronic patients Free episomic forms and replicative intermediates o f HBV were Found inter mittently and,in one o£ these,Northern blot of PMBCs RNA detected specific HBV RNA transcripts. In conclusion: 1. Specific HBV transcripts in RMBCs of a patient with chronic active hepatitis is the First confirmation in humans of what has already been observed in the woodchuck model. 2. As no PMBCs specific HBV signals were shown in the acute hepatitis group, if involvement occurs, it may be a very early Feature of HBV infection. 3. The presence of HBV in P ~ C s seems to be transient in time and the events able to activate the replication are still unknown. This study was supported by Associazione Italiana per la Ricerca sul Cancro AIRC.
$59
SEROLOGIC PROFILE OF HEPATITIS D VIRUS (HDV) RELATED FULMINANT HEPATITIS. A.Mas, M. Buti, J. Costa, J.M.Sanchez-Tapias, R. Esteban, J.Guardia and J.Rod~s. Liver Unit, Hospital Cllnico and Dept. of Internal Medicine, Hospital Valle de Hebron, Barcelona, Spain.
To assess the diagnostlc value of HDV serum markers in fulminant hepatitis D a longitudinal study of the serologic p r o f i l e of 27 patients who were referred for HBsAg or IgM anti-HBc positive fulminant hepatic f a i l u r e has been performed. SerumHDAg, t o t a l a n t i HD, IgM anti-HD and RNA-VHD were examined every two to three days during the phase of severe l i v e r f a i l u r e and weekly thereafter in survivors. A single HDV immunological marker or a combination of markers was detected in the f i r s t serum sample from 18 patients. HBsAg was present in a l l and 14 of them also had IgM a n t i HBc. HDV markers were never detected in the nine remalning patients. HDAg alone was detected in six cases, HDAg and anti-HD in five ( t o t a l anti-HD in one and both t o t a l and IgM anti-HD in four) and a n t i - delta alone in seven ( t o t a l anti-HD in two and both t o t a l and IgM anti-HD in f i v e ) . Thus, isolated IgM anti-HD was never observed. HDV-RNA was detected only in three patients with concomitant HD antigenemia. All patients with HDAg in serum were admitted less than ten days after the beginning of symptoms. Among patients with HDAg alone, seroconversion to both t o t a l and IgM anti-HD occurred within a few days in three cases whlle isolated HDAg remained in three further patients who died shortly. In patients i n i t i a l l y showing both HDAg and anti-HD, the antigen became undetectable a few days l a t e r . Changes of the i n i t i a l serological pattern were not observed in patients with isolated anti-HD in the i n i t i a l sample. The precedent observations indicate that testing for both HDAg and t o t a l anti-HD is mandatory for the diagnosis of fulminant hepatitis D while IgM anti-HD and HDV-RNA have l l t t l e diagnostic value in this sltuation.
114
HBV DNA IN PERIPHERAL MONONUCLEARBLOOD CELLS: RELATIONSHIP WITH VIRAL REPLICATION M.Melegari,C.Pasquinelli,M.Solieri,F.Manenti,E.Villa Chair of Gastroenterology, University of Modena, Italy.
Hepatitis B virus (HBV) infection of peripheral mononuclear blood cells (PMBCS) has been recently reported as a Feature occurring in all stages of liver disease. Moreover, in the related woodchuck hepatitis virus model specific transcripts have been shown in FMBCs and in the spleen of chronically infected animals, suggesting that a complete replicative cycle can take place in htese cells. We therefore investigated 30 patients with acute type B hepatitis (6/30 HBV DNA+, 14/30 HBeAg+) and a smaller group of 5 patients with chronic active hepatitis HBeAg+, HBV DNA+,followed up at monthly intervals. Southern and Northern blot analysis of PMBCs were performed utilizing a Full lenght HBV probe. No HBV specific sequences in PMBCs were displayed in the acute hepatitis group either in Southern or Northern blot, although the sensitivity of our method enabled us to detect as little as 1 pg of cloned HBV insert. On the contrary,in 5/5 chronic patients Free episomic forms and replicative intermediates o f HBV were Found inter mittently and,in one o£ these,Northern blot of PMBCs RNA detected specific HBV RNA transcripts. In conclusion: 1. Specific HBV transcripts in RMBCs of a patient with chronic active hepatitis is the First confirmation in humans of what has already been observed in the woodchuck model. 2. As no PMBCs specific HBV signals were shown in the acute hepatitis group, if involvement occurs, it may be a very early Feature of HBV infection. 3. The presence of HBV in P ~ C s seems to be transient in time and the events able to activate the replication are still unknown. This study was supported by Associazione Italiana per la Ricerca sul Cancro AIRC.
$59