Lymphocytes were cultured for 3 to 5 days with hrIL-2 and Con-A, counted, and prepared at a density of 4.0 x 10e6 cells/ml of media. Approximately 10e7 cells were seeded into eight separate 50 ml centrifuge tubes, and cells were treated with polybrene for 1 hour at 37oC.
The cells were washed and infected with 16 TCID50 of USgaB01 for 2 hours, or mock infected with "spent" media. The cells were washed, re-suspended in 10 ml of fresh media, and aliquots were cultured for 1, 2, 4, and 24 hours. Total RNA was isolated at each time
point for microarray analysis. Individual aliquots of lymphocytes were infected on 3 separate
INSTRUMENT(S): (Make:Axon Instruments,Model:GenePix 4000A)
ORGANISM(S): Felis catus
SUBMITTER: Ryan J O Dowling
The Journal of general virology 20050801 Pt 8
Infection of cats with Feline immunodeficiency virus (FIV) is an important model for understanding comparative lentivirus biology. In vivo, FIV infects lymphocytes and monocyte/macrophages, but in vitro infection is commonly investigated in epithelial Crandell-Reese Feline Kidney (CRFK) cells. In this study, the transcriptional responses of CRFK cells and primary lymphocytes to infection with FIV 34TF, a cloned subtype A virus, and FIV USgaB01, a biological subtype B isolate, were determined. Re ...[more]