Figure 4

ImmTAV redirection of CD8⁺ T cells to HIV-infected primary CD4⁺ T cells. (a) Gag p24 total corrected cellular fluorescence (TCCF; as measured from confocal microscopy images) of activated or resting HIV-infected primary CD4⁺ T cells in conjugates with ImmTAV-redirected HIV-naïve donor CD8⁺ T cells (bottom); each dot represents a conjugate (n > 20/condition). Horizontal lines indicate median value. Groups were analysed by Mann Whitney test. Representative images of conjugates with activated infected CD4⁺ T cell targets shown as Gag expression in resting infected cells was not visible (top). (b) Confocal microscopy images of Zap70 localisation to the IS (examples of activated and resting, top), % Zap70 localisation at IS (bottom left) or cytoplasmic vs. membrane distribution (bottom right). (c) Confocal microscopy of α-tubulin (MTOC shown with white arrow; examples of activated and resting, left) and distance from MTOC to the synapse (µM, right). (d) Confocal microscopy of perforin localisation (docked example, left) and % of conjugates with distal, dispersed or docked perforin (right) in the CD8⁺ T cell. All synapse markers: at least 8 conjugates analysed per condition. Red = p24*, magenta = CD8 on DIC image, green = synapse molecule. For (b)–(d), horizontal lines indicate mean values. Groups were analysed by unpaired t test. *As p24 intensity in resting infected cells is very low and thus difficult to visualize, only DIC & CD8 signal images are shown.