Distribution of viral RNA in mouse tissues during acute phase of coxsackievirus B5 infection.
OBJECTIVE
To investigate histopathological changes and distribution of coxsackievirus B5 (CVB5) RNA in mouse heart, liver, and pancreas during the acute phase of infection.


METHODS
C3H/HeJ male mice, aged 3-4 weeks, were inoculated intraperitoneally with 5 x 10(5) plaque-forming units of CVB5 and sacrificed at 1, 2, 3, 4, 7 and 10 days postinfection (p.i.). Inflammation of the heart, liver, and pancreatic tissue sections was evaluated by hematoxylin and eosin staining, and virus was detected using antibody to viral coat protein VP1. A quantitative real-time RT-PCR method, using primers and probe targeted to the highly conserved sequences in the 5'-untranslated region of the virus, was used to evaluate the kinetics of CVB5 RNA during the development of myocarditis or pancreatitis.


RESULTS
Marginal inflammatory changes were observed in the heart tissues although viral RNA was constantly present between 1 and 10 days p.i., peaking at 4 days p.i. The pancreatic tissues displayed massive lymphocyte infiltration and loss of acinar cells at day 4 p.i. and viral RNA was detected between 1 and 10 days p.i., peaking at 2-3 days p.i. In the liver, viral RNA was detected between 1 and 7 days. No mortality was observed.


CONCLUSIONS
CVB5 induced acute pancreatitis without subsequent development of myocarditis. Clearance of CVB5 RNA from the pancreas and heart was slower than clearance from the liver. Our real-time RT-PCR method, which is more sensitive than conventional plaque assay, may provide valuable insight into viral RNA kinetics during CVB5 infection.
Moon
Mi Sun
MS
Department of Microbiology, University of Ulsan College of Medicine, Seoul, Korea.
Joo
Chul Hyun
CH
Hwang
In Seok
IS
Ye
Jeong Sook
JS
Jun
Eun Jung
EJ
Lee
Hui Sun
HS
Kim
Donghou
D
Lee
Min-Jae
MJ
Lee
Heuiran
H
Kim
Yoo Kyum
YK
D000208:Acute Disease; D000818:Animals; D003384:Coxsackievirus Infections / Q000473:pathology* / Q000821:virology*; D029822:Enterovirus B, Human / Q000302:isolation & purification / Q000502:physiology*; D006321:Heart / Q000821:virology; D008099:Liver / Q000473:pathology / Q000821:virology; D008297:Male; D051379:Mice; D009205:Myocarditis / Q000473:pathology / Q000821:virology; D009206:Myocardium / Q000473:pathology; D010179:Pancreas / Q000473:pathology / Q000821:virology; D010195:Pancreatitis / Q000473:pathology / Q000821:virology; D012367:RNA, Viral / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D010948:Viral Plaque Assay
D012367:RNA, Viral