Key Points
- To fractionate and evaluate the immunoreactivity of hypertonic salt-extracted cardiac antigens in a murine model of Coxsackievirus B3-induced myocarditis.
- Extracted proteins from heart tissues of adolescent CD-1 mice using hypertonic salt and separated them into two fractions using Sephadex G-100 column chromatography.
- Tested cell-mediated immune responses via an agarose droplet cell migration inhibition assay using peritoneal exudate cells from infected, immunized, and unimmunized mice, with liver and spleen fractions as controls.
- Assessed the viral neutralizing activity of anti-fraction antibodies via plaque reduction assays and analyzed protein composition using 10% SDS-polyacrylamide gel electrophoresis.
- Fraction I contained antigenic activity that significantly inhibited migration of peritoneal exudate cells from CVB3-infected mice, whereas Fraction II displayed no inhibitory activity.
- Fraction I derived from control liver or spleen tissues failed to inhibit immune cell migration, and unimmunized mouse cells showed no migration inhibition in response to Fraction I.
- Antibodies raised against Fraction I and Fraction II did not neutralize CVB3m virus in plaque reduction assays, and gel electrophoresis of Fraction I resolved multiple distinct protein bands.
Fraction I of KCl-extracted cardiac antigens from CVB3-infected mice exhibits specific antigenic immunoreactive activity in vitro.