Apc cy7 anti human cd4 clone rpa t4

Example 26

To test the effect of CD137 agonistic antibodies on human cells in vivo, human PBMCs (7×10⁶) were intravenously injected to immunocompromised NSG mice (NOD.Cg-Prkdc^scid Il2^rgtm1Wjl/SzJ; Jackson Laboratory; Cat #005557). The mice were randomized to groups of 8 and received CD137 antibodies (200 μg/mouse) or vehicle control on days 0, 7 and 14. Peripheral blood from each mouse was collected on days 10, 20 and 29 for determination of human CD45+(FITC anti-human CD45 clone 11130; BioLegend; Cat #304038), CD8+(ALEXA FLUOR® 647 anti-human CD8a clone HIT8a; BioLegend; Cat #300918), and CD4+(APC-Cy7 anti-human CD4 clone RPA-T4; Bd; Cat #557871) engraftment using flow cytometry.

FIGS. 34A-34C show overall increase in numbers of hCD45+ cells and systemic hyper expansion of human CD8+ T cells in mice that received mAb4 at the expense of human CD4+ T cells. Notably, mAb1 did not induce over activation of human T cells. Reduced potential of mAb1 to activate human T cells in the periphery might contribute to reduced potential for toxicity.

Example 26

To test the effect of CD137 agonistic antibodies on human cells in vivo, human PBMCs (7×10⁶) were intravenously injected to immunocompromised NSG mice (NOD.Cg-Prkdc^scid Il2rg^tm1wjl/SzJ; Jackson Laboratory; Cat #005557). The mice were randomized to groups of 8 and received CD137 antibodies (200 μg/mouse) or vehicle control on days 0, 7 and 14. Peripheral blood from each mouse was collected on days 10, 20 and 29 for determination of human CD45 (FITC anti-human CD45 clone H130; BioLegend: Cat #304038). CD8⁺ (Alexa Fluor 647 anti-human CD8a clone HIT8a; BioLegend; Cat #300918), and CD4⁺ (APC-Cy7 anti-human CD4 clone RPA-T4; Bd; Cat #557871) engraftment using flow cytometry.

FIGS. 34A-34C show overall increase in numbers of hCD45⁺ cells and systemic hyper expansion of human CD8+ T cells in mice that received mAb4 at the expense of human CD4+ T cells. Notably, mAb1 did not induce over activation of human T cells. Reduced potential of mAb1 to activate human T cells in the periphery might contribute to reduced potential for toxicity.