In situ hybridization studies of hepatitis A viral RNA in patients with acute hepatitis A.
In situ hybridization with oligonucleotide probes has been used to localise hepatitis A virus RNA genomic sequences in formalin-fixed and routinely processed human liver biopsies from three patients. Using radiolabelled Sulphur-35 antisense probes, viral genomic sequences were found in all three cases, but signal intensity was greatest in cases 1 and 2 with fulminant hepatitis, and was minimal in the third case of resolving hepatitis biopsied 2 months after acute illness. Localisation showed the viral RNA to be present in hepatocytes, sinusoidal cells and inflammatory cells in and around the portal tracts. Both cases showed signal in similar cell types, but the distribution of staining was predominantly periportal in case 1, whereas lobular staining was more apparent in case 2. Hybridization with sense polarity probes failed to detect any evidence of replicative intermediates of antigenomic viral RNA. The presence of hepatitis A RNA in phagocytic cells was confirmed using immunohistochemistry for a macrophage marker, CD68, combined with in situ hybridization. In all cases the signal was predominantly cytoplasmic, and this was confirmed with the use of tritiated probes.
Department of Histopathology, St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London, UK.
M
M
Taylor
R D
RD
Goldin
S
S
Ladva
P J
PJ
Scheuer
H C
HC
Thomas
D000208:Acute Disease; D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D006506:Hepatitis A / Q000235:genetics* / Q000473:pathology; D017957:Hepatitis A Virus, Human / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008099:Liver / Q000737:chemistry / Q000473:pathology; D008264:Macrophages / Q000737:chemistry / Q000473:pathology; D008297:Male; D008875:Middle Aged; D015345:Oligonucleotide Probes; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics*; D000097382:CD68 Molecule
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D015345:Oligonucleotide Probes; D012367:RNA, Viral; C067980:CD68 antigen, human; D000097382:CD68 Molecule