J Bacteriol Virol.  2008 Dec;38(4):239-247. 10.4167/jbv.2008.38.4.239.

Expression of Plus- and Minus-strand Viral RNA in Coxsackievirus B3-Infected A/J Mice

• Hwang IS
• Jun EJ
• Ye JS
• Joo CH
• Lee H
• Kim YK

• KMID: 1483965
• DOI: http://doi.org/10.4167/jbv.2008.38.4.239

Abstract

In order to investigate the implication of viral replication in acute, subacute, and chronic infections of coxsackievirus B3 (CVB3), we examined the histopathological changes and plus- and minus-strand viral RNA dynamics in heart, pancreas, brain, and liver of CVB3-infected A/J mice. Mice were inoculated intraperitoneally with CVB3 and sacrificed on 1, 2, 3, 4, 7, 10, 14, 21, 30, 60, and 90 days post infection (p.i.). Plus- and minus-strand viral RNAs in the organs were quantitated and the organs were additionally evaluated histopathologically for inflammation. No inflammatory infiltrates were observed in the liver, brain, and heart. In contrast, massive lymphocyte infiltration and fat replacement were shown in the pancreas with loss of acinar cells. Both plus- and minus-strand viral RNA levels were detected by 21 days p.i. in heart, 90 days p.i. in pancreas, 4 days p.i. in liver, and 10 days p.i. in brain. The plus-strand RNA was found at least fifty fold higher than the minus-strand RNA by 4 days p.i. in heart and pancreas and by 3 days p.i. in liver. The plus- to minus-strand RNA ratio in brain was found less than 1:20. Our data indicate that viral replication was actively occurred in heart, pancreas, and liver during acute CVB3 infection, whereas viral replication was limited in brain. Furthermore, chronic persistent viral RNA was observed in pancreas. In conclusion, CVB3 at low dose of virus induces severe pancreatitis but marginal or no inflammatory changes in the heart, liver, and brain.

Keyword

Coxsackievirus B3; Pancreatitis; Viral RNA dynamics; A/J mice

MeSH Terms

Acinar Cells
Animals
Brain
Heart
Inflammation
Liver
Lymphocytes
Mice
Pancreas
Pancreatitis
RNA
RNA, Viral
Viruses
RNA
RNA, Viral

Figure

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  Figure 1. Hematoxylin and eosin (H&E)-stained paraffin-embedded murine heart tissue sections (3-μm) obtained on 1, 2, 3, 4, 7, 10, 14, 21, 30, 60, and 90 days p.i. of CVB3. No inflammatory infiltrates were observed during experiments except 1 or 2 mice at 10 and 30 days p.i. (×20).

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  Figure 2. Hematoxylin and eosin (H&E)-stained paraffin-embedded murine pancreatic tissue sections (3-μm) obtained on 1, 2, 3, 4, 7, 10, 14, 21, 30, 60, and 90 days p.i. The affected pancreata displayed massive lymphocyte infiltration and loss of acinar cells, which peaked on 4 and 7 days p.i. of CVB3. Restoration of pancreatic tissues was observed between days 10 and 14 p.i., thereafter tissues were redestructed.

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  Figure 3. Increase of inflammatory cell infiltration and acinar cell destruction were found at acute phase in pancreatic tissue infected with CVB3. Fat replacement was prominent at chronic phase.

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  Figure 4. Standard curve used to determine CVB3 RNA concentration in tissue samples.

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  Figure 5. Copy number of plus- and minus-strand viral RNAs in heart, pancreas, liver and brain obtained from A/J mice infected with CVB3. Both plus- and minus-strand viral RNAs were peaked on 2 days p.i., then decreased in all tissue samples. (A) No viral RNA was detected after 30 days p.i. (B) Both plus- and minus-strand viral RNAs were detected until 90 days p.i. Both strand viral RNAs were detected until 4 days p.i. (C) and 10 days, respectively, except days 7 p.i. (D). The numbers above histogram are the ratio of plus to minus strand viral RNA.

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  Figure 6. The ratio of plus- to minus-strand RNA in various organs obtained from A/J mice infected with CVB3. In heart, pancreas and liver in acute phase, high ratio (>20) of plus- to minus-viral RNA means the presence of active viral replication. Lower ratio was shown in brain.

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