Experimental hepatitis A virus (HAV) infection in cynomolgus monkeys (Macaca fascicularis): evidence of active extrahepatic site of HAV replication.
This work studied the replication sites of hepatitis A virus (HAV) in cynomolgus monkeys (Macaca fascicularis) after intravenous inoculation. The cynomolgus monkeys were inoculated with the Brazilian hepatitis A virus strain (HAF-203). Monkeys were euthanized on days 15, 30, 45 and 60 postinoculation (pi). Liver samples, submandibular salivary gland, mesenteric lymph node and tonsils were removed for virological and pathological evaluation. Immunofluorescence analyses on liver and salivary gland sections using confocal laser scanning microscopy revealed the presence of HAV antigen (HAV Ag). The presence of HAV genome was monitored by real-time PCR. The HAV RNA was detected at 7 days postinoculation (dpi), concomitantly in serum, saliva and faeces. The highest HAV viral load was observed in faeces at 15 dpi (10(5) copies/ml), followed by serum viral load of 10(4) copies/ml at 20 dpi and saliva viral load of 10(3 )copies/ml at 7 dpi. The animals showed first histological and biochemical signs of hepatitis at 15 dpi. The HAV antigen (Ag) was present from day 7 until day 60 pi in the liver and salivary glands. The HAV replicative intermediate was also detected in the liver (4.5 x 10(4) copies/mg), salivary glands (1.9 x 10(3) copies/mg), tonsils (4.2 x 10(1) copies/mg) and lymph nodes (3.4 x 10(1) copies/mg). Our data demonstrated that the salivary gland as an extrahepatic site of early HAV replication could create a potential risk of saliva transmitted infection. In addition, the cynomolgus monkey was confirmed as a suitable model to study the pathogenesis of HAV human infection.
Amado
Luciane A
LA
Laboratório de Desenvolvimento Tecnológico em Virologia, Instituto Oswaldo Cruz/Fiocruz, Rio de Janeiro, Brazil. l_amado@ioc.fiocruz.br
Marchevsky
Renato S
RS
de Paula
Vanessa S
VS
Hooper
Cleber
C
Freire
Marcos da S
Mda S
Gaspar
Ana Maria C
AM
Pinto
Marcelo A
MA
D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D004195:Disease Models, Animal; D005243:Feces / Q000821:virology; D005455:Fluorescent Antibody Technique; D006506:Hepatitis A / Q000175:diagnosis* / Q000473:pathology / Q000635:transmission; D035922:Hepatitis A Antibodies / Q000097:blood; D035921:Hepatitis A Antigens / Q000302:isolation & purification; D030041:Hepatitis A virus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D007275:Injections, Intravenous; D008099:Liver / Q000201:enzymology / Q000821:virology; D008198:Lymph Nodes / Q000821:virology; D008252:Macaca fascicularis; D008297:Male; D018613:Microscopy, Confocal; D014066:Palatine Tonsil / Q000821:virology; D012367:RNA, Viral / Q000097:blood / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012463:Saliva / Q000821:virology; D012469:Salivary Glands / Q000821:virology; D013997:Time Factors; D019562:Viral Load; D014779:Virus Replication
D035922:Hepatitis A Antibodies; D035921:Hepatitis A Antigens; D012367:RNA, Viral; D000410:Alanine Transaminase