Conversion of a tumor-binding peptide identified by phage display to a functional chimeric T cell antigen receptor.
Adoptive transfer of ex vivo expanded tumor-specific T cells is a promising therapeutic modality for promoting or augmenting antitumor immunity. Several groups, including ours, are developing antigen receptor gene transfer strategies as a means of generating effector cells for adoptive therapy. Chimeric antigen receptors (CARs) have been described that use single-chain antibodies or cytokine ligands as tumor targeting domains. Here, we describe the capacity of a tumor-binding peptide identified by phage display combinatorial library screening to serve as a CAR targeting domain. A phage library-selected high-affinity 12-mer peptide (Bpep) specific for alpha(v) beta(6) integrin (alpha v beta6) was chosen for these studies. Primary human T cells were genetically modified to express the Bpep-CAR consisting of an alpha v beta6-specific peptide and human IgG4 hinge-Fc extracellular domain fused to the cytoplasmic tail of CD3-zeta. T cell expression of the Bpep-CAR was assessed by Western blot analysis, and trafficking of the Bpep-CAR to the cell surface was demonstrated by flow cytometry. Functionally, Bpep-CAR redirected cytotoxic T lymphocytes specifically kill integrin alpha v beta6+ ovarian tumor targets, and are activated for interferon gamma secretion. Our data suggest that large new repertoires of tumor-specific T cell antigen receptor transgenes might be available through merging combinatorial peptide libraries with CAR construct design.
Pameijer
C R J
CR
Division of General and Oncologic Surgery, Stony Brook University Hospital, Stony Brook, NY, USA.
Navanjo
A
A
Meechoovet
B
B
Wagner
J R
JR
Aguilar
B
B
Wright
C L
CL
Chang
W-C
WC
Brown
C E
CE
Jensen
M C
MC
D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000378:metabolism; D002460:Cell Line; D005260:Female; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism
D000951:Antigens, Neoplasm; D016023:Integrins; D009842:Oligopeptides; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; C073878:integrin alphavbeta6