Detection of lymphocytes expressing hepatitis B virus in peripheral blood from HBsAg positive and negative patients by “in situ” hybridization

CYCLOSPORINi IMMUNOSUPPRESSIVE TREATMENT AND LIVER FUNCTION IN HEART TRANSPLANTATION (HTx) B. Gulbis, M. Adler~ H.A. Ooms, J.M. Desmet, J.L. Leclerc, G. Primo Department of Clinical Chemistry,XGastroenterology and Cardiac S~-rge--~, H6pital Erasme - ULB, Brussels, I070 - Route de Lennik, 808. The problem of cyclosporin (CsA) hepatotoxicity has been reported with variable incidence and has not been studied systematically. This study includes 15 patients undergoing HTx and treated by CsA, azathioprine and corticosteroids. Liver function was assessed before and during the postoperative course (l month) of HTx by 5 biological tests: serum total bile acids (BA), alkaline phosphatase (ALP), glutamyl transf~rase ( g t ) , total b i l i r u b i n ( B i l i T.), and aminopyrine breath test (ABT). These tests permit to classify the patients in 3 groups before HTx: normal (n = 7) (Group I ) , mild altered (n = 4) (Group I I ) and severe altered (n = 4) l i v e r function (Group I l l ) . A 24h CsA oral kinetic (300 mgr) versus placebo was performed in group I; serum CsA levels and the same biological tests except ABT were followed during these kinetics. During the postoperative period: BA and gt levels incresed significantly only in group I l l (2 to II times upper l i m i t of normal.p O.Ol). The area under the curve of serum CsA levels and BA levels were calculated; the ratio AUCRJAVCrAwere correlated into the groups and significantly elevatedl in group I l l compared to ~oup ~ and I I (p O.OOl). During kinetics, only BA levels increased in every patients without any elevation during the placebo study. All these data have suggested a direct effect of CsA on BA "secretion". BA seemed in this work a more sensitive indicator of CsA effect on l i v e r than "classical" biological l i v e r tests.

DETECTION OF LYMPHOCYTESEXPRESSING HEPATITIS B VIRUS IN PERIPHERAL BLOOD FROMHBsAg POSITIVE AND NEGATIVE PATIENTS BY "IN SITU" HYBRIDIZATION. M. HADCHOUELI. C. PASQUINELLI2, R.N. HUGONI, C. BRECHOT2, O. BERNARDi. I)--FNS'ERM U 5-~--,H6pital de Bic~tre, 94275~ Le KremT~-nC~-Tc~tre Cedex, France. 2) UREG, INSERM U 163, I n s t i t u t Pasteur, 75724 Paris Cedex 15, France. By using "in situ" hybridization methodology we have examined lymphocytes from peripheral blood from HBsAg positive and negative patients with l i v e r disease for the presence of hepatitis B virus RNA. Mononuclear cells preparations were hybridized with a S3s or H3 labeled HBV specific DNA probe and exposed to autoradiographic emulsion. 44 subjects were studied (26 children, 18 adults). The results : Serological status

Nb

Ag HBs+, Ag HBe+ (17) Ag HBs+, Ag HBe- (18) Anti-HBs+ (3) HCA sero-negative (2) Controls (4)

Serum HBV/DNA 14/17 0/8 0/3 0/2 0/4

Lymphocytes HBV/RNA 10/17 8/18 I/3 2/2 0/4

The frequency of labeled cells varied from one patient to another from 1 to 10 %. These results demonstrate the presence of viral RNA in mononuclear cells and indicate that the DNA detected so far in human mononuclear cells is transcriptionnally active.

$128