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Phycoerythrin conjugated Goat anti Mouse IgG1 antibody (STAR132PE) used for the detection of bound antibody on lymphoblastoid cell lines by flow cytometry.
Estimation of ΔBNLF2a and wild-type LCLs expressing lytic antigens; recognition by immediate early antigen-specific CD8+ T cells. The proportion of LCLs spontaneously reactivating into lytic cycle was assessed by intracellular BZLF1 staining and analysis by flow cytometry, with representative examples shown for LCLs derived from two different donors: (A) donor 1 and (B) donor 2. Immediate early lytic cycle CD8+ T cell recognition of wild-type (wt), ΔBNLF2a (Δ2a) and ΔBZLF1 (ΔBZ) LCLs using HLA-B*0801-restricted RAK (BZLF1) clones against appropriately HLA matched donor 1 and 2 LCLs (C and D respectively) was measured by IFNγ ELISA. Results using wild-type or ΔBNLF2a cells diluted with ΔBZLF1 cells as appropriate are shown, where arrows indicate equivalent numbers of lytic antigen expressing cells. Experiments were also conducted using HLA-C*0202-restricted IACP (BRLF1) clones against donor 3 LCLs (E), and HLA-B*4501-restricted AEN (BRLF1) clones against donor 4 LCLs (F). For donor 4, both the wild-type and ΔBNLF2a LCLs were diluted with ΔBZLF1 LCL (wild-type-LCL titration data not shown). Data are represented as mean+/-SEM.
From: Croft NP, Shannon-Lowe C, Bell AI, Horst D, Kremmer E, et al. (2009) Stage-Specific Inhibition of MHC Class I Presentation by the Epstein-Barr Virus BNLF2a Protein during Virus Lytic Cycle. PLoS Pathog 5(6): e1000490.