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Immunohistochemical detection of HIV1 in liver cells
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I M M U N O H I S T O C H E M I C A L D E T E C T I O N O F H I V I IN L I V E R CELLS
C. Housset*, Q, Boucher**. P.M: Girard**. J. Leibowitch***. A.G. Saimot**. C. Br~chot*. C. Marche** * Unit6 d'H6patologie, HOpital La~nnec, INSERM U - 7 5 C H U Necker ** H6pital Claude Bernard *** H6pital R a y m o n d Poincare, Paris
A n i m m u n o h i s t o c h e m i c a l method was used to investigate the possibility of liver cell infection by HIVI. Liver samples from 17 patients seropositive for HIVI (AIDS : 15, A R C : 1, unclassified : 1) were obtained by percutaneous biopsy (12 cases : 10 p r e - , 2 post m o r t e m ) surgical biopsy (1 case) or at autopsy (4 cases). Cryostat sections were labelled with a three-stage indirect i m m u n o p e r o x i d a s e technique. Monoclonal antibodies against HIVI gag protein p24/p55 (Abbott) and CD4 : leu 3a (Becton Dickinson) were used as primary antisera. L y m p h nodes f r o m infected individuals and liver sections f r o m seronegative subjects were respectively used as positive and negative controls. Monoclonal anti p24/p55 produced liver cell staining in 7 out of 17 interpretable samples. It consisted in 4 cases (2 p r e - , 2 p o s t - m o r t e m ) of a d i f f u s e cytoplasmic staining of n u m e r o u s K u p f f e r cells. In 2 f u r t h e r autopsy s p e c i m e n s and 1 p r e - m o r t e m percutaneous biopsy, granular staining of scattered cells was shown : positive cells included sinusoidal cells as well as, mononuclear cells present in dense i n f l a m m a t o r y infiltrates. In addition, in all cases the CD4 molecule was localized, using leu 3a antibody, on endothelial sinusoisal cells. Conclusion : These results suggest HIV infection of non p a r e n c h y m a l liver cells. CD4 molecule was identified on endothelial cells. T h e staining obtained with the anti-P24 antibody is mainly localized in mononuclear i n f l a m m a t o r y and sinusoidal cells w h i c h seem to be respectively histiocytes and K u p f f e r cells. T h u s , HIV infection of m o n o c y t e - m a c r o p h a g e cell type m i g h t be involved in some liver pathological lesions in AIDS, such as sinusoidal abnormalities.
76
LINKAGE OF DNA MARKERS ON CHROMOSOME13 WITH WILSON DISEASE RHJ Houwen, H Scheffer*, GJ Te Meerman*, P vd V l i e s * , HSA Heymans, CHCMBuys* Department of Pediatrics and *Department of Human Genetics, University of Groningen, The Netherlands.
Wilson disease (WD) is an autosomal recessive disorder, characterized by massive copper deposition in the l i v e r , the basal ganglia and in other organs. The basic defect in WD is s t i l l unknown. Recently a close linkage has been reported between the gene f o r WD and the esterase-D locus in some Middle East f a m i l i e s . (Lod-score = 5.16;recombination f r a c t i o n 0 . 0 5 ) These r e s u l t s , pointing to a l o c a l i s a t i o n of the WD-gene in band q14 of chromosome 13, were confirmed in Sardinian WD-families, but not in Canadian WD-families. Therefore we investigated a number of Dutch-Caucasian f a m i l i e s with WD. Linkage studies were carried out using ESD, a cDNA probe of esterase-D. Our f i r s t results are from t h i r t e e n large f a m i l i e s , with 25 children c l a s s i f i e d as diseased and 31 as healthy, based on serum copper, ceruloplasmin and 24 hr urinary copper excretion. In our f a m i l i e s the lod score f o r ESD was only 0.09 at a recombination f r a c t i o n of 0.30. These results do not yet support a close linkage between ESD, and WD. These results are in concordance with the data from Canada, but not with the Sardinian and Middle-East data . Therefore we suggest the existence of at least two populations with.respect to the l o c a l i s a t i o n of the WD gene: a Caucasian and a Middle-East/Sardinian population.
$40
I M M U N O H I S T O C H E M I C A L D E T E C T I O N O F H I V I IN L I V E R CELLS
C. Housset*, Q, Boucher**. P.M: Girard**. J. Leibowitch***. A.G. Saimot**. C. Br~chot*. C. Marche** * Unit6 d'H6patologie, HOpital La~nnec, INSERM U - 7 5 C H U Necker ** H6pital Claude Bernard *** H6pital R a y m o n d Poincare, Paris
A n i m m u n o h i s t o c h e m i c a l method was used to investigate the possibility of liver cell infection by HIVI. Liver samples from 17 patients seropositive for HIVI (AIDS : 15, A R C : 1, unclassified : 1) were obtained by percutaneous biopsy (12 cases : 10 p r e - , 2 post m o r t e m ) surgical biopsy (1 case) or at autopsy (4 cases). Cryostat sections were labelled with a three-stage indirect i m m u n o p e r o x i d a s e technique. Monoclonal antibodies against HIVI gag protein p24/p55 (Abbott) and CD4 : leu 3a (Becton Dickinson) were used as primary antisera. L y m p h nodes f r o m infected individuals and liver sections f r o m seronegative subjects were respectively used as positive and negative controls. Monoclonal anti p24/p55 produced liver cell staining in 7 out of 17 interpretable samples. It consisted in 4 cases (2 p r e - , 2 p o s t - m o r t e m ) of a d i f f u s e cytoplasmic staining of n u m e r o u s K u p f f e r cells. In 2 f u r t h e r autopsy s p e c i m e n s and 1 p r e - m o r t e m percutaneous biopsy, granular staining of scattered cells was shown : positive cells included sinusoidal cells as well as, mononuclear cells present in dense i n f l a m m a t o r y infiltrates. In addition, in all cases the CD4 molecule was localized, using leu 3a antibody, on endothelial sinusoisal cells. Conclusion : These results suggest HIV infection of non p a r e n c h y m a l liver cells. CD4 molecule was identified on endothelial cells. T h e staining obtained with the anti-P24 antibody is mainly localized in mononuclear i n f l a m m a t o r y and sinusoidal cells w h i c h seem to be respectively histiocytes and K u p f f e r cells. T h u s , HIV infection of m o n o c y t e - m a c r o p h a g e cell type m i g h t be involved in some liver pathological lesions in AIDS, such as sinusoidal abnormalities.
76
LINKAGE OF DNA MARKERS ON CHROMOSOME13 WITH WILSON DISEASE RHJ Houwen, H Scheffer*, GJ Te Meerman*, P vd V l i e s * , HSA Heymans, CHCMBuys* Department of Pediatrics and *Department of Human Genetics, University of Groningen, The Netherlands.
Wilson disease (WD) is an autosomal recessive disorder, characterized by massive copper deposition in the l i v e r , the basal ganglia and in other organs. The basic defect in WD is s t i l l unknown. Recently a close linkage has been reported between the gene f o r WD and the esterase-D locus in some Middle East f a m i l i e s . (Lod-score = 5.16;recombination f r a c t i o n 0 . 0 5 ) These r e s u l t s , pointing to a l o c a l i s a t i o n of the WD-gene in band q14 of chromosome 13, were confirmed in Sardinian WD-families, but not in Canadian WD-families. Therefore we investigated a number of Dutch-Caucasian f a m i l i e s with WD. Linkage studies were carried out using ESD, a cDNA probe of esterase-D. Our f i r s t results are from t h i r t e e n large f a m i l i e s , with 25 children c l a s s i f i e d as diseased and 31 as healthy, based on serum copper, ceruloplasmin and 24 hr urinary copper excretion. In our f a m i l i e s the lod score f o r ESD was only 0.09 at a recombination f r a c t i o n of 0.30. These results do not yet support a close linkage between ESD, and WD. These results are in concordance with the data from Canada, but not with the Sardinian and Middle-East data . Therefore we suggest the existence of at least two populations with.respect to the l o c a l i s a t i o n of the WD gene: a Caucasian and a Middle-East/Sardinian population.
$40