Cardiac Fibroblasts Aggravate Viral Myocarditis: Cell Specific Coxsackievirus B3 Replication

Figure 5

Virus clearance or persistence in macrophages infected with high or low MOI of CVB3. Murine macrophages (RAW cells) were infected with 5 MOI or 0.5 MOI of CVB3. The copy number of the viral RNA genome (+) strand and the copy number of the intermediate (−) strand needed for virus replication were determined using specific primers for reverse transcription followed by TaqMan analysis. (a) Copy numbers of both CVB3 strands using different MOI 6 hours and 24 hours after infection. (b) Ratio between copy numbers after 24 hours after infection compared to 6 hours after CVB3 infection. Ratios lower than 1 show virus elimination. (c) To determine gene expression of macrophages infected with 5 or 0.5 MOI of CVB3 total RNA was reverse transcribed using random primer and gene expression of TNF-α, MCP-1, and TGF-β was determined 6 hours after infection. (d) 24 hours after infection with CVB3 cells were washed to remove cell debris and then total RNA was isolated from the remaining cells. The isolated amount of total RNA is plotted and revealed a decrease due to virus infection followed by cell lysis in cardiac fibroblasts and cardiomyocytes but not in macrophages. (a) Significantly different versus copy numbers 6 hours after infection; . Data are plotted as mean ± SEM. (b) Significantly different versus 5 MOI; . Ratios are plotted as box plots with median. (c) Significantly different versus noninfected control, ; ; . Data are plotted as mean ± SEM.

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