Coxsackievirus B1-induced murine myositis: no evidence for viral persistence

Abstract

The persistence of coxsackievirus B1 in the muscles of mice with coxsackievirus Bl-induced chronic myositis was investigated. Neonatal CD1 Swiss mice were inoculated with a myositis-causing variant of coxsackievirus B1 (Tucson strain). Hamstring muscle samples of diseased mice obtained at various times after inoculation were examined for the presence of infectious virus, viral RNA and histological abnormalities. Viral RNA was detected up to 4 weeks after initiation of infection, whereas virus could be isolated from hamstring muscles for up to 2 weeks. Thereafter no sign of infection was demonstrated although histological abnormalities remained present for the entire observation period of 16 weeks. That viral RNA was detectable for only 2 weeks after tissues became negative for infectious virus suggests that the infection slowly waned rather than the viral RNA persisting. Hence, it is concluded that coxsackievirus B1 plays an essential role in the initiation of myositis but not in the maintenance of the chronic phase.

Predominant right leg dysfunction without asymmetric muscle inflammation in CD1 Swiss mice with coxsackievirus B1-induced myositis

“…Muscles were paraffin-embedded and sectioned at 6 /¿m. In a previous study we found that most extensive inflammatory and myopathic changes occurred in hamstring muscles, in agreement with previous reports (19,38) and with the fact that these muscles were the most atrophic on macroscopy (27,32). W e also dem on strated that the histological changes (m ononuclear cell infiltra tion, muscle fiber necrosis, central nuclei, fiber atrophy) were evenly distributed over the w hole length of a muscle (2 0 ), and that cross section areas of ham string muscles at 2 weeks did not differ significantly from those at 4 weeks (19).…”

Kissing of the two predominant hairpin loops in the coxsackie B virus 3' untranslated region is the essential structural feature of the origin of replication required for negative-strand RNA synthesis

“…Total RNA was isolated from 100 l of cellular lysates obtained from the 8-h time point of the growth curve, using a single extraction procedure with guanidinium thiocyanate-phenol-chloroform (9). Mutated RNA was PCR amplified, using a poly(T) primer and a primer located in the 3D coding region (5Ј-GTTGTTTGACCCTCCCCGCG-3Ј; nt 7241 to 7260) as described previously (48). The resultant 179-bp reverse transcriptase PCR products were purified by low-melting-point agarose gel electrophoresis and sequenced as described above.…”

Genetic analysis of a hydrophobic domain of coxsackie B3 virus protein 2B: a moderate degree of hydrophobicity is required for a cis-acting function in viral RNA synthesis

“…RNA was isolated from 100 l of virus stock by a single extraction procedure with guanidinium thiocyanate-phenol-chloroform according to the method described by Chomczynski and Sacchi (11). Reverse transcription of RNA and amplification of cDNA by PCR with forward primer 5Ј-GCAATGGAACAGGGAGTGAAG GACTATGTGGA-3Ј (nt 3733 to 3765) and reverse primer 5Ј-TTGGGATGGC GCGCTCTGCTC-3Ј (nt 4231 to 4251) were performed as described previously (42). The resulting 519-bp PCR products were purified by low-melting-point agarose gel electrophoresis.…”