Cloning and Expression of Human Membrane-Bound and Soluble Engineered T Cell Receptors for Immunotherapy

[摘要] We report here the design and construction of several gene vectorsfor expression in mammalian cells of membrane-bound and solublehuman T cell receptors (TR). We designed a vector(TR-ALPHA-IRES-TR-BETA pEF4) that encodes high-level expression ofthe full-length TR on the surface of T cells. Furthermore, weengineered TR that does not require the presence of endogenous CD3molecules for surface expression and thus expression is notlimited to T cells. We also constructed a vector encoding asingle-chain TR (scTR) as a fusion protein ofV-ALPHA-V-BETA-C-BETA with CD3Z. Since it is encoded and expressedas a single molecule, this scTR is well suited for gene therapy.Lastly, we successfully used a mammalian expression vector forgeneration of soluble human TR. The approaches we used here formanipulation of a human tumor-specific TR can be useful for otherinvestigators interested in TR-based immunotherapy.

[学科分类] 基础医学