Mimotopes for Alloreactive and Conventional T Cells in a Peptide–MHC Display Library
Figure 5
Recovery of IAb-p3K Virus-Infected Cells with Fluorescent αβTCR
(A) Sf9 cells were infected with a mixture of virus, 99% of which encoded a control protein (a TCR β chain linked to the gp64 transmembrane/cytoplasmic tail) and 1% of which encoded IAb-p3K. After 3 d, the infected cells were analyzed as in Figure 3A for binding fluorescent αβTCR from YAe-62. The 1% of the infected cells with the brightest fluorescence was sorted (high sort, 15,700 cells). As a control, a similar number of cells that fluoresced as dully as the background fluorescence were also sorted (low sort).
(B) The sorted cells were incubated with fresh Sf9 insect cells to allow propagation of the viruses and production of new stocks. The stocks were used to infect new Sf9 cells, and after 3 d the analysis of αβTCR binding was repeated.