Telbivudine in Chronic Lymphocytic Myocarditis and Human Parvovirus B19 Transcriptional Activity

Figure 1. Impact of telbivudine on parvovirus B19‐infected human microvascular endothelial cells. (A) Experimental design: human microvascular endothelial cells (HMEC)‐1 were 24 h after plating, infected with 10 000 viral particles/cell with/out 10 ng/mL of telbivudine, and collected after 30 min for the analysis of tenascin‐C and TGF‐β1 mRNA expression, after 24 h for the analysis of apoptosis, and after 72 h for the anaylsis of endothelial‐to‐mesenchymal transition (EndMT). Bar graphs represent the mean ± SEM of (B) Annexin V/7AAD‐ cells (% gated), (C) α‐smooth muscle actin (SMA)+/VE‐cadherin‐ cells (% gated), (D) tenascin‐C, (E) transforming growth factor (TGF)‐β, and (F) interferon (IFN)‐β mRNA expression, as indicated, with n = 4–6/group and *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Figure 2. Impact of telbivudine on parvovirus B19 (B19V) copy number and expression in B19V‐infected human microvascular endothelial cells. (A) Experimental design: human microvascular endothelial cells (HMEC‐1) were 24 h after plating, infected with 10 000 viral particles/cell with/out 10 ng/mL of telbivudine, and collected after 1, 24, or 48 h for the analysis of B19V DNA copy number and mRNA expression. Bar graphs represent the mean ± SEM of (B) B19V copy number and (C) B19V cDNA, 1, 24, and 48 h post‐B19V infection in HMEC‐1 with n = 5–6/group and *P < 0.05 and ****P < 0.0001, (D) B19V copy number (n = 4/group), and (E) B19V cDNA expression (n = 6/group) 24 h post‐infection with/out telbivudine treatment.

Figure 3. Impact of telbivudine on parvovirus B19 (B19V) copy number and expression in B19V‐infected UT7/EPO S1 cells. (A) Experimental design: UT7/EPO S1 cells were 24 h after plating, infected with 10 000 viral particles/cell with/out 10 ng/mL of telbivudine, and collected after 1 and 24 h for the analysis of B19V DNA copy number and mRNA expression. Bar graphs represent the mean ± SEM of (B) B19V copy number and (C) B19V cDNA, 1 and 24 h post‐B19V infection with/out telbivudine treatment in UT7/EPO S1 cells with n = 4–6/group and **P < 0.005, ***P < 0.0005, and ****P < 0.0001.

Figure 4. Pleiotropic effects of telbivudine. The pleiotropic effects of telbivudine were evaluated in tumour necrosis factor (TNF)‐α‐stimulated human microvascular endothelial cells (HMEC)‐1 (A‐C), angiotensin (Ang) II‐stimulated HL‐1 cells (D‐F), and transforming growth factor (TGF)‐β1‐stressed fibroblasts (G‐H). (A) Experimental design: HMEC‐1 were 24 h after plating, stimulated with/out 10 ng/mL of TNF‐α in the presence or absence of 10 ng/mL of telbivudine, and collected after 24 h for the analysis of apoptosis, or stained with Sirius Red and crystal violet for the quantification of collagen deposition. Bar graphs represent the mean ± SEM of (B) Annexin V/7AAD‐ cells (% gated) (n = 5–6/group), (C) the ratio of the absorbances at 495 to 405 nm (n = 10–12/group). (D) Experimental design: HL‐1 were 24 h after plating, stimulated with/out 1 μM Ang II in the presence or absence of 10 ng/mL of telbivudine, and collected after 24 h for the analysis of reactive oxygen species (ROS) and myosin. Bar graphs represent the mean ± SEM of (E) DCF (% gated cells) (n = 5–6/group), (F) myosin (% gated cells) (n = 6/group). (G) Experimental design: fibroblasts were 24 h after plating, stimulated…

Figure 5. In situ tissue typing of endomyocardial biopsies in parvovirus B19‐positive myocarditis patients before (mRNA+) and after telbivudine treatment (mRNA−) via imaging mass spectrometry. (A) Probabilistic latent semantic analysis could distinguish the protein signatures of the B19V mRNA+ (red) and B19V mRNA− (blue) patients (Patients 3 and 4 of Table 1). (B) Peptides from tenascin‐C (TNC; 1740 Da) show significantly lower intensities (receiver operating characteristic > 0.72; P < 0.001) in B19 mRNA− compared with B19V mRNA+ endomyocardial biopsy.

Figure 6. Study design of the PreTopic study. EF, ejection fraction; EMB, endomyocardial biopsy.