Active Coxsackieviral B Infection Is Associated With Disruption of Dystrophin in Endomyocardial Tissue of Patients Who Died Suddenly of Acute Myocardial Infarction
Laurent Andréoletti, MD, PhD*,*,
Lydie Ventéo, MSci ,
Fatima Douche-Aourik, PhD ,
Frédéric Canas, MD,
Geoffroy Lorin de la Grandmaison, MD, PhD ,
Jérôme Jacques, MSci*,
Hélène Moret, PhD*,
Nicolas Jovenin, MD||,
Jean-François Mosnier, MD, PhD¶,
Mathieu Matta, MSci*,
Sébastien Duband, MD#,
Michel Pluot, MD,
Bruno Pozzetto, MD, PhD and
Thomas Bourlet, MD, PhD
* Laboratoire de Virologie Médicale et Moléculaire et Faculté de Médecine (EA-3798), Reims, France
Service d'Anatomie Pathologique, Centre Hospitalier Universitaire, Reims, France
Laboratoire de Bactériologie-Virologie, Centre Hospitalier Universitaire, Saint-Etienne, France
Service d'Anatomie Pathologique et de Médecine Légale, Hôpital Raymond Poincaré, Garches, France
|| Département d'Information Médicale, Centre Hospitalier Universitaire et Faculté de Médecine de Reims, Reims, France
¶ Service d'Anatomie Pathologique, Centre Hospitalier Universitaire, Nantes, France
# Service d'Anatomie Pathologique, Centre Hospitalier Universitaire, Saint-Etienne, France
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Figure 1 Immunostaining of VP1 in Heart Tissue of Patients With MI and Control Patients
Immunostaining (red) of capsid viral protein 1 (VP1) in heart tissue of a patient with an inaugural acute myocardial infarction (MI) (A), a patient with acute MI in the context of chronic coronary heart disease (B), and a negative healthy heart patient (C). The sections were counterstained with hematoxylin. Magnification x400.
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Figure 2 Gel Electrophoresis of PCR Products
(A) Polymerase chain reaction (PCR) products generated by nested reverse transcriptase (RT)-PCR using primers located in the 5'-noncoding region of the enterovirus genome (360 bp). Lanes 1 to 4 correspond to patients who died of an inaugural acute myocardial infarction (MI); lanes 5 to 8 correspond to patients with a previous history of chronic coronary disease and who died of an acute MI; P1 = positive control obtained by RT-PCR amplification of positive ribonucleic acid (RNA) strands (2 x 1010 copies), which were transcribed from a coxsackievirus B3 (CV-B3) complementary (c) deoxyribonucleic acid (DNA) sequence; P2 = positive control obtained by direct PCR amplification of single-strand copies of CV-B3 cDNA; NC1 = negative control of RT assay and nested PCR amplification; NC2 = negative control (sterile water) of the first PCR amplification assay; NC3 = negative control of the second PCR amplification assay; M = molecular size marker (100-bp DNA ladder). (B) PCR products generated by RT-PCR using primers located in the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) housekeeping gene (190 bp). Lanes 1 to 8 correspond to a positive GAPDH messenger RNA detection in cardiac tissues from patients 1 to 8. NC4 = RT-PCR negative control (sterile water); M = molecular size marker (100-bp DNA ladder).
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Figure 3 Alignment of Amino Acid Sequences of Partial Capsid Viral Protein of Cardiac EV Isolates From 8 Patients Who Died of Acute MI
Amino acid sequences of partial capsid viral protein region of cardiac enterovirus (EV) isolates from 8 patients who died of acute myocardial infarction (MI), and of reference coxsackievirus B (CV-B) sequences retrieved from GenBank (M16560, AF085363, M16572, and X05690 for CV-B1, -B2, -B3, and -B4, respectively). The alignments were performed with reference to the CV-B2 sequence. The homology was >85% between sequences of strains from patients 1 to 4 and CV-B2 and from patients 5 to 8 and CV-B3. Differences are printed in full. Dash = identical amino acid; dot = amino acid deletion; star = undetermined amino acid.
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Figure 4 Immunohistochemical Analyses Showing the Sarcolemmal Staining Pattern for Dystrophin Using Antidystrophin Antibody
Dystrophin pattern was conserved in cardiac tissue sections of (A) healthy heart controls (immunostaining in red) and of (B) enterovirus (EV)-negative patients who died of acute myocardial infarction (immunostaining in red). Example of focal areas of myocardium displaying a loss of the sarcolemmal staining pattern in cardiac tissue sections of EV-positive patients (B and E, arrows). In several study cases, immunohistochemical analyses of serial cardiac sections demonstrated that there was focal disruption of the sarcolemmal organization of dystrophin (F, arrows) in the same areas of cardiac tissue that were positive for the EV capsid viral protein 1 (C, arrows) (immunostaining in brown). The sections were counterstained with hematoxylin. Magnification: x400 (A and B); x1,000 (C to F).
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