Generating B7-H3-Targeted CAR T-Cells

We generated the B7-H3-targeted V_HH-based CAR and 376.96(scFv)-based CAR lentiviral vector following the design principle of the CAR construct published in our previous study^{21 (link)}. Briefly, individual V_HH (B12/C4/G8) fragments or scFv of 376.96 mAb were respectively subcloned into a CAR construct (pMH338, pMH340, pMH339, and pMH372). The CAR-expressing lentivirus was produced as described previously^{21 (link)}. PBMCs isolated from four healthy donors (#076, #1, #2, and #3) were stimulated for 24 hours using anti-CD3/anti-CD28 antibody-coated beads (Invitrogen) at a bead: cell ratio of 2:1 according to manufacturer’s instructions in the presence of IL-2.

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Li D., Wang R., Liang T., Ren H., Park C., Tai C.H., Ni W., Zhou J., Mackay S., Edmondson E., Khan J., Croix B.S, & Ho M. (2023). Camel nanobody-based B7-H3 CAR-T cells show high efficacy against large solid tumours. Nature Communications, 14, 5920.

Publication 2023

anti-CD3/anti-CD28 antibody-coated beads

Anti antibody Cell Donors Lentivirus Vector

Corresponding Organization :

Other organizations : Center for Cancer Research, National Cancer Institute, IsoPlexis (United States), Frederick National Laboratory for Cancer Research

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Variable analysis

independent variables

V_H_H (B12/C4/G8) fragments
ScFv of 376.96 mAb

dependent variables

CAR-expressing lentivirus

control variables

PBMCs isolated from four healthy donors (#076, #1, #2, and #3)
Anti-CD3/anti-CD28 antibody-coated beads (Invitrogen) at a bead: cell ratio of 2:1

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