title: Cloning and expression of human membrane-bound and soluble engineered T cell receptors for immunotherapy.
abstract: We report here the design and construction of several gene vectors for expression in mammalian cells of membrane-bound and soluble human T cell receptors (TR). We designed a vector (TR-ALPHA-IRES-TR-BETA pEF4) that encodes high-level expression of the full-length TR on the surface of T cells. Furthermore, we engineered TR that does not require the presence of endogenous CD3 molecules for surface expression and thus expression is not limited to T cells. We also constructed a vector encoding a single-chain TR (scTR) as a fusion protein of V-ALPHA-V-BETA-C-BETA with CD3Z. Since it is encoded and expressed as a single molecule, this scTR is well suited for gene therapy. Lastly, we successfully used a mammalian expression vector for generation of soluble human TR. The approaches we used here for manipulation of a human tumor-specific TR can be useful for other investigators interested in TR-based immunotherapy.
authors.affiliation: Department of Immunology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors.forename: Nehad M
authors.initials: NM
authors.lastname: Alajez
authors.forename: Saman
authors.initials: S
authors.lastname: Eghtesad
authors.forename: Olivera J
authors.initials: OJ
authors.lastname: Finn