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1、 An Experimental Model for Dilated Cardiomyopathy after Rabbit Coronavirus Infection Author(s): Lorraine K. Alexander, J. David Small, Suzanne Edwards and Ralph S. Baric Source: The Journal of Infectious Diseases, Vol. 166, No. 5 (Nov., 1992), pp. 978-985 Published by: Oxford University Press Stable
2、 URL: http:/www.jstor.org/stable/30113367 Accessed: 17-09-2016 05:54 UTC JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivi
3、ty and facilitate new forms of scholarship. For more information about JSTOR, please contact supportjstor.org. Your use of the JSTOR archive indicates your acceptance of the Terms revised 11 June 1992. Financial support: National Institutes of Health (AI-23946); American Heart Association (AHA 90111
4、2; Established Investigator Award AHA 890192 to R.S.B.). Reprints or correspondence: Dr. Ralph S. Baric, Department of Epidemi- ology, School of Public Health, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7400. The Journal of Infectious Diseases 1992;166:978-85 ? 1992 by The Un
5、iversity of Chicago. All rights reserved. 0022-1899/92/6605-0005501.00 strated after coxsackievirus B or encephalomyocarditis virus infection in mice. In these models, viral infection results in myocarditis and CHF, with a significant percentage of survi- vors developing DCM at a later stage in life
6、 14-16. A rabbit model for coronavirus-induced heart disease has been described 17, 18. Infection with rabbit coronavirus (RbCV) resulted in virus-induced myocarditis and CHF with a mortality rate of 60. Morphologic and pathologic evi- dence indicates that a significant percentage of these animals w
7、ere dying from heart failure 17. We determined whether survivors of RbCV infection would develop DCM. Materials and Methods Animals and virus. Male New Zealand White rabbits weigh- ing 2.5-3.0 kg were purchased from commercial suppliers (Franklin Rabbitry, Wake Forest, NC, or Robinson Services, Wins
8、ton-Salem, NC). The animals were housed individually at room temperature (210C) and given rabbit diet (Agway; Grand- ville Milling, Creedmoor, NC) and water ad libitum. RbCV stocks were obtained from moribund animals when peak titers were present at 4 days after infection 17. Virus stocks were dilut
9、ed to 103 -104 rabbit ID5o per milliliter and stored at -140oC. Animals were inoculated either intravenously via the marginal ear vein or occasionally intramuscularly in the thigh muscle with 0.2 mL of the 103 -104 RID50 virus stock. No differ- ences were observed in day of death, clinical signs of
10、infection, or histologic findings between the routes of inoculation. Ani- mals were observed daily for signs of infection, which included weight loss, dullness of the sclera, hyphema, and severe conges- tion of the conjunctivae and irides. For virus isolation from the heart muscle, 7 survivors at 30
11、- 111 days after infection were sacrificed by intravenous injection of 50 mg/kg sodium pentobarbital. The hearts were removed and flushed extensively or perfused with PBS, pH 7.0. The apex of the ventricles was removed, snap frozen in liquid nitrogen, then stored at -140oC until assayed. One gram of
12、 tissue was minced and homogenized on ice in 4 mL of PBS using a manual This content downloaded from 130.63.180.147 on Sat, 17 Sep 2016 05:54:24 UTC All use subject to http:/about.jstor.org/terms JID 1992; 166 (November) Coronavirus: Dilated Cardiomyopathy tissue grinder. Large pieces of tissue were
13、 removed by centrifuga- tion at 12,000 g for 10 min in an Eppendorf centrifuge (Fisher Scientific, Norcross, GA) at 4CC. Rabbits were inoculated with 0.5 mL of the undiluted heart homogenate supernatant, then observed daily for 14 days for signs of viral infection. After 21 days, rabbits were challe
14、nged with 0.2 mL of stock serum at 103 ? 104 RIDso/mL and observed daily for 14 days for signs of infection. Pathology. Survivors of RbCV infection and uninfected control animals were sacrificed as previously described. Body weights were obtained to the nearest 0.10 kg. The heart was removed, separa
15、ted from the pericardial sac, and flushed with PBS. The heart was weighed to the nearest 0.1 g; then the chambers were filled and fixed with 10% phosphate-buffered for- malin. The heart was sectioned transversely at the widest dimen- sions of the ventricles. Four paraffin-embedded 6-jum sections wer
16、e cut at 150-m intervals and stained with hematoxylin-eo- sin. Additional heart sections were also stained with von Kossas method or Masson trichrome stain. Sections of lung, liver, and spleen were also removed, fixed in 10 phosphate-buffered for- malin, and stained with hematoxylin-eosin. Morphometric studies. To determine the degree of myocyte hypertrophy, myocytes in the right and left ventricles and the interventricular septum were measured by using a software m
