Figure 4

Panel A: Proliferation Index (PI) time course comparison in three donors' (A, B, and C) PBLs cultured with either PHA or B7 surface-engineered HIV-1 based particles. For PHA (black-filled bars), the proliferation value in the presence of PHA (i.e. Donor-A, 6 hour timepoint = 10,900 relative fluorescent units) is divided by untreated cultures not exposed to PHA (i.e. Donor-A, 6 hour timepoint = 2,500 relative fluorescent units); for B7 (gray-filled bars) the proliferation value in the presence of B7 surface-engineered particles (i.e. Donor-A, 6 hour timepoint = 32,500 relative fluorescent units) is divided by the proliferation value observed with non-engineered HIV-based particles (i.e. Donor-A, 6 hour timepoint = 2,500 relative fluorescent). The time course shown is 4, 6, 8, 12, and 18 days for B7; 4, 6, 8, and 12 days for PHA. Particle preparations used in this panel were PEG-concentrated (200× for HIV; 25× for HSV) and inactivated to render them non-infectious. Panel B: Proliferation Index (PI) time course comparison in Donor-F PBLs cultured with HSV-2 based surface-engineered particles. For PHA (black-filled bars), the proliferation value in the presence of PHA (i.e. 8 hour timepoint = 9,000 relative fluorescent units) is divided by cultures not exposed to PHA (i.e. 8 hour timepoint = 2,600 relative fluorescent units); for AntiCD3 surface-engineered particles (tightly packed horizontal hatched gray bars), the proliferation value in the presence of AntiCD3 (i.e. 8 hour timepoint = 15,000 relative fluorescent units) is divided by the proliferation value observed with non-engineered HSV-based particles (i.e. 8 hour timepoint = 2,300 relative fluorescent units); for B7 surface-engineered particles (horizontal hatched gray bars), the proliferation value in the presence of B7 (i.e. 8 hour timepoint = 29,000 relative fluorescent units) is divided by the proliferation value observed with non-engineered HSV-based particles (i.e. 8 hour timepoint = 2,300 relative fluorescent units); for B7+antiCD3 surface-engineered particles (horizontal hatched bars), the proliferation value in the presence of B7+antiCD3 (i.e. 8 hour timepoint = 28,000 relative fluorescent units) is divided by the proliferation value observed with non-engineered HSV-based particles (i.e. 8 hour timepoint = 2,300 relative fluorescent units). Particle preparations used in this panel were from conditioned media and inactivated to render them non-infectious. Panel C: Proliferation Index (PI) time course comparison in Donor-F PBLs cultured with HSV-2 based surface-engineered particles. For PHA (black-filled bars), the proliferation value in the presence of PHA (i.e. 8 hour timepoint = 4,200 relative fluorescent units) is divided by cultures not exposed to PHA (i.e. 8 hour timepoint = 1,200 relative fluorescent units); for Heat-Inactivated B7+antiCD3 surface-engineered particles (tightly packed horizontal hatched gray lines), the proliferation value in the presence of heat-inactivated surface-engineered particles (i.e. 8 hour timepoint = 7,300 relative fluorescent units) is divided by the proliferation value observed with heat-inactivated non-engineered HSV-based particles (i.e. 8 hour timepoint = 6,500 relative fluorescent units); for Conditioned Media B7+antiCD3 (horizontal hatched bars), the proliferation value in the presence of conditioned media from surface-engineered particles (i.e. 8 hour timepoint = 30,000 relative fluorescent units) is divided by the proliferation value observed in conditioned media from non-engineered HSV-based particles (i.e. 8 hour timepoint = 2,700 relative fluorescent units); for Lyophilized room temperature stored B7+antiCD3 (checker bars), the proliferation value in the presence of the lyophilized surface-engineered particles (i.e. 8 hour timepoint = 28,000 relative fluorescent units) is divided by the proliferation value observed with lyophilized non-engineered HSV-based particles (i.e. 8 hour timepoint = 2,300 relative fluorescent units). The above data was obtained using Donor-F PBLs. PEG-concentrated B7+antiCD3 (brick bars) proliferation value was compared to Conditioned Media B7+antiCD3 (horizontal hatched bars) in Donor-H PBLs. The remaining PI values are calculated in a similar fashion. Almost identical "background" values are observed for non-PHA exposed and non-engineered particles in Donors-A, -B, -C, -F, and -H cultures. Actual induced values can be calculated by multiplying the PI value by the "background" value. Particle preparations used in this panel unless otherwise identified were from conditioned media and inactivated to render them non-infectious.