Intracellular viral localization in murine coxsackievirus-B3 myocarditis. Ultrastructural study by electron microscopic in situ hybridization.
Group B Coxsackieviruses are a common cause of myocarditis. To detect the viral genome and its localization in the myocardium, we examined C3H/He mice with Coxsackievirus B3 (CVB3) myocarditis on days 5, 8, and 14 after inoculation by the reverse transcriptase polymerase chain reaction and by in situ hybridization. Sense and antisense CVB3 RNA were detected in the myocardium of all mice up to day 14 by reverse transcriptase polymerase chain reaction. Light microscopic in situ hybridization with a cDNA probe for CVB3 showed clusters of positive signals in the areas of myocardial necrosis and cell infiltration. With electron microscopic in situ hybridization, CVB3 RNA was detected in the cytoplasm of cardiocytes, between the myofibrils, near the mitochondria, and in tubular or vesicular structures. Viral RNA was also detected in necrotic debris, in the cytoplasm of macrophages, and in the cytoplasm of interstitial fibroblasts. These findings suggest that CVB3 RNA is replicated in the cytoplasm of cardiocytes, transferred into tubular or vesicular structures, released into the interstitium, and phagocytosed by macrophages. Some positive signals were also detected in the cytoplasm of cardiocytes showing close contact with infiltrating lymphocytes, suggesting that the lymphocytes recognized virus-infected cardiocytes and caused cell-mediated immune cardiocyte damage.
Ukimura
A
A
Department of Internal Medicine, Osaka Medical College, Japan.
Deguchi
H
H
Kitaura
Y
Y
Fujioka
S
S
Hirasawa
M
M
Kawamura
K
K
Hirai
K
K
D000818:Animals; D003384:Coxsackievirus Infections / Q000473:pathology / Q000821:virology*; D029822:Enterovirus B, Human / Q000235:genetics / Q000302:isolation & purification*; D006321:Heart / Q000821:virology*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008214:Lymphocytes / Q000166:cytology / Q000648:ultrastructure; D008264:Macrophages / Q000166:cytology / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008854:Microscopy, Electron; D009205:Myocarditis / Q000175:diagnosis / Q000378:metabolism / Q000821:virology*; D009206:Myocardium / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D012367:RNA, Viral / Q000032:analysis*; D013997:Time Factors
D012367:RNA, Viral