EBV Virus Production for Rabbit Inoculation

EBV producing B95-8 cells were cultured at 37 °C in RPMI (GIBCO) supplemented with 10% Fetal Bovine Serum (FBS) (GIBCO), 50 µg/ml gentamycin (Hyclone), 1% antibiotic and antimycotic solution (Santa Cruz) and 1X glutamine (GIBCO), as previously described^{24 (link)}. Cells were grown to a density of approximately 6.5 × 10⁶cells/ml and then incubated at 30ºC for 24 h to promote viral production. The culture supernatant was centrifuged to remove cell debris and filtered using a 0.2 µm filter. Freshly filtered culture supernatant containing the virus was then used for intravenous inoculation of rabbits. Quantitative real-time PCR (qRT-PCR) was used to determine EBV copy number in the inoculum, using Namalwa cell line DNA as standards^{55 (link)}.