Expansion of Functional Polyclonal Human T-regulatory Cell Lines Using Artificial Antigen Presenting Cells and Rapamycin. (B159)

Abstract

Abstract T-regulatory CD4+CD25+ T lymphocytes play a central role in regulating immune response and preventing autoimmunity. There is a broad spectrum of potential clinical applications of expanded T-regs including but not limited to treatments of graft versus host disease, transplant rejection, and type I diabetes. K562 cell line with stable expression of lentivirus-introduced CD64 and CD86 markers (KT64/86) and loaded with anti-CD3 antibodies was used to expand human CD4+CD25 high T lymphocytes. Though in several occasions stimulation of CD4+CD25high cells with KT64/86 only resulted in production of functional T-regulatory cells, only when antibiotic rapamycin was added to the cultures, functional Tregs were produced in 100% of stimulations. These cells were also active in vivo in reversing GVHD in NOG mice. Total CD4 lymphocytes grown in the presence of rapamycin though suppressive in vitro were not able to reverse GVHD in mice. Substitution of CD86 with CD80 resulted in a less potent Tregs. Addition of certain members of TNF receptor family such as CD70, OX40 ligand or 4-1BB ligand to KT64/86 cells, had different effects on expansion of Treg cell lines. While CD70 addition did not show any impact, 4-1BB and OX-40 ligands in most cases stimulated growth of T-regulatory cell lines but at the same time in most cases also attenuated their activity. The project is supported by JDRF Collaborative Center for Cell Therapy grant to C.J.