Cases/Abstracts

Submitted by Purva Nagarajan, University of Pennsylvania 

Generation of swine gamma delta CAR T cell lines

Purva Nagarajan, Lidia Flor Cuenca, Jack Swain, Hannah Thomas, Nimisha Pattada, Raimon

Duran-Struuck

Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania,

Philadelphia, PA.

Chimeric antigen receptor (CAR) T cell therapies use T cells engineered to target antigens

expressed in cancer cells. Research has been developed to optimize this technology using animal

models such as rodents. Unfortunately, rodent models have demonstrated poor clinical predictive

value, limiting the translatability to man. Swine share many anatomical, physiological, and

immune characteristics with humans, making them ideal for clinical translation. Gamma delta T

cells (GDCs) are a subset of CD3+ cells that can be found in the peripheral blood and mucosal

tissues and have strong anti-tumor properties and no MHC restriction. This makes them an

attractive cell type as there is minimal risk for inducing graft-versus-host disease compared to

alpha-beta T cells. We sought to take advantage of both CAR technology and GDC

characteristics to generate a large animal CAR GDC. Using swine peripheral blood mononuclear

cells, we isolated and expanded GDCs for transduction with a characterized CAR molecule.

Optimization of expansion techniques are being studied using zoledronic acid, IL-2, and artificial

antigen presenting cells (aAPCs) based on a K562 cells line expressing human CD86 and CD64

loaded with anti-swine CD3. To date, using 1uM ZA, 1000 IU/IL2 and a 10:1 aAPC:GDC ration

lead to a >4-fold expansion of absolute GDCs within 8 days. Flow cytometry data suggests that

the majority of these GDCs are CD4- and CD8-. Lentiviral transduction of GDCs with an anti-

HLA A2 CAR, characterized previously in human and swine T cell effectors, yielded up to a

24.8% CAR transduction making them a viable new cell line for clinical application upon further

refinement of the protocol.

Research grant: ITMAT

Student support: Internal funds