Activation and Expansion of T-Cells Using an Engineered Multivalent Signaling Platform as a Research Tool

Abstract

Provided are a system and methods for selectively inducing expansion of a population of T cells in the absence of exogenous growth factors, such as lymphokines, and accessory cells for research purposes. The cell based expansion system and methods permit the long-term growth of CTLs, preferably human CTLs. In addition, T cell proliferation can be induced without the need for antigen, thus providing an expanded T cell population that is polyclonal with respect to antigen reactivity. Further provided are methods for using the system and methods to screen and identify antigens related to specific diseases or conditions, tumors, autoimmune disorders, or an infectious disease or pathogen, and to identify target molecule for research purposes, or for developing a vaccine based thereon.

1 . A method of preparing an aAPC comprising: selecting a population of K562 cells and engineering the cells to express Fcγ receptor CD32, thereby creating K32 cells; transfecting the K32 cells with the co-stimulatory ligand; cloning the K32 cells to permit exogenous loading of the anti-CD3 or anti-CD28 antibodies, or a combination thereof, and then loading of the anti-CD3 or anti-CD28 antibodies, or a combination thereof. 
     
     
         2 . The method of  claim 2 , wherein the ligand is 4-1BB (4-1BBL).