Postnatal Persistent Infection with Classical Swine Fever Virus and Its Immunological Implications

Muñoz-González, Sara; Ruggli, Nicolas; Rosell, Rosa; Pérez, Lester Josué; Frías-Leuporeau, Maria Teresa; Fraile, Lorenzo; Montoya, Maria; Cordoba, Lorena; Domingo, Mariano; Ehrensperger, Felix; Summerfield, Artur; Ganges, Llilianne

The 6D10 + cell subsets were sorted using a live sterile cell sorting system (FACSAria, Beckton Dickinson, San Jose, California, USA). To obtain 6D10 + cells, 24 x10 6 BMHCs were incubated with 6D10 hybridoma supernatant for 1 h on ice, washed with PBS containing 2% FBS, and incubated with R-phycoerythrin conjugate goat (Fab 0 ) 2 anti-mouse Ig (Dako, Denmark). Single cell sorting was performed in using purity precision mode, with a 70 μm nozzle. The fluorescence reading was performed upon excitation with a 488 nm argon laser. The 6D10 + and 6D10 − cells were more than 95% pure by flow cytometry, and a total of 6701444 6D-10 + cells were recovered with 97% efficiency. The presence of CSFV RNA in both types of recovered cells was analysed by RT-PCR [36] .