This study investigates the use of bispecific antibodies and effector T cells in targeting tumor cells in a mouse model. The approach aims to enhance T cell infiltration and tumor cell apoptosis through engineered antibody interactions.
Take an anesthetized, immunocompromised mouse bearing a subcutaneous tumor xenograft that overexpresses a tumor-associated antigen or TAA.
Retroorbitally inject effector T cells expressing the luciferase enzyme.
Subcutaneously administer interleukin 2, a cytokine supporting effector T cell survival.
Retroorbitally inject bispecific antibodies engineered to contain binding sites for T and tumor cells. Allow the mouse to recover.
The antibodies and T cells reach the tumor via circulation.
The antibodies bind to CD3 on the T cells and TAA on the tumor cells, inducing the effector T cells to release toxins that initiate apoptosis in the tumor cells and release chemokines that recruit more T cells to the tumor.
Anesthetize the mouse, retroorbitally inject a luciferase substrate, and proceed to in vivo imaging.
Luciferase inside the T cells oxidizes the substrate, emitting bioluminescence.
Observe the bioluminescence over time. The magnitude of the signal correlates with the extent of T cell infiltration within the tumor.
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.
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