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

After anesthetizing the mouse using a 26-gauge needle, inject 20 million T cells in 100 microliters of media retro-orbitally. Administer 1,000 units of recombinant IL-2 subcutaneously to support T cell survival in vivo. Then, administer the bispecific antibody retro-orbitally or intraperitoneally.

For in vivo imaging, administer 100 microliters of three milligrams of D-luciferin by retro-orbital injection with a 26-gauge needle in an anesthetized mouse. To capture images, place the mouse in the light-tight chamber of the imager, such that the flank bearing the xenograft is facing up toward the camera. Using the acquisition control panel, select luminescent, photograph, and overlay. Set the exposure time to auto, the binning to medium, and F-stop to one, and acquire images.

After the first image, set the exposure time to match that automatically calculated for the first image so that subsequent images may be compared directly. Take another set of images with the mouse supine to evaluate T cell presence in the lungs.