BioMed Research International / 2014 / Article / Fig 4

Research Article

Combined Cytolytic Effects of a Vaccinia Virus Encoding a Single Chain Trimer of MHC-I with a Tax-Epitope and Tax-Specific CTLs on HTLV-I-Infected Cells in a Rat Model

Figure 4

Characterization of m8Δ expressing SCTs. (a) Diagram of SCTs encoding Tax180–188 or NLEnv371–379 linked to m and RT1. molecules with two linkers. L1, linker 1; L2, linker 2; TM, transmembrane region; Cyto, cytoplasmic region. ((b) and (c)) Whole cell extracts were isolated from RK13 cells infected with indicated clones of m8Δ/RT1AlSCTax180L (b) or RK13 cells infected with m8Δ (lane 1) or m8Δ/RT1AlSCNLEnv371L (lane 2) (c) and 50 μg of each lysate was subjected to Western blotting analysis for the expression of the SCT of MHC-I proteins. Arrows indicate the SCT of MHC-I proteins detected by an anti-rat MHC-I antibody. Molecular weight markers are indicated (kDa) on the left margin. (d) RK13 cells were exposed to m8Δ/RT1AlSCTax180L, m8Δ/RT1AlSCNLEnv371L, or PBS for 2 hrs. After extensive wash, the cells were cocultivated with 4O1/C8 for indicated days. Production of IFN-γ in the supernatants was measured by ELISA. The data represent the mean ± the SD of triplicate wells. Statistical significance was determined as . Similar results were obtained in two independent experiments.