An In Vivo Technique to Track Bispecific Antibody-Induced T Cell Trafficking to Tumors

Abstract

Source: Espinosa-Cotton, M., et al. Tracking Bispecific Antibody-Induced T Cell Trafficking Using Luciferase-Transduced Human T Cells. J. Vis. Exp. (2023).

This video demonstrates a method for tracking bispecific antibody-induced T cell trafficking to tumors in a mouse model. Luciferase-transduced effector T cells are injected into a mouse with a subcutaneous tumor cell xenograft. Bispecific antibodies are then introduced, binding to T cells and the tumor cells. The localization of the T cells is tracked using bioluminescent imaging, with the magnitude of the signal indicating the extent of T cell infiltration into the tumor.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Transfection of 293T cells with luciferase and harvest of viral supernatant

1. Culture of 293T cells
   1. Prepare media by adding the following to a liter of Dulbecco's modified Eagle medium (DMEM) each: 110 mL of heat-inactivated fetal bovine serum (FBS), 11 mL of penicillin-streptomycin.
   2. Thaw 5 x 10⁶ 293T cells and transfer them into a T175 flask with the media prepared as described above.
   3. Split the 293T cells 1:10 every 3 ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
293T cells	ATCC	CRL-11268
BSA	Sigma Aldrich	A7030-10G
CD3/CD28 beads	Gibco (ThermoFisher)	11161D
D-Luciferin, Potassium Salt	Goldbio	LUCK-1G
DMEM	Gibco (ThermoFisher)	11965092
DNA in vitro transfection reagent (polyjet)	SignaGen Laboratories	SL100688
EDTA	Sigma Aldrich	E9884-100G
FBS	Gibco (ThermoFisher)	10437028
Gag/pol plasmid	Addgene	14887
GFP plasmid	Addgene	11150-DNA.cg
Penicilin-Streptomycin	Gibco (ThermoFisher)	15140122
Recombinant human IL-2	R&D Systems	202-IL-010/CF
Retronectin	Takara	T100B
Trypsin	Gibco (ThermoFisher)	25-300-120
VSV-G plasmid	Addgene	8454