T cell growth control using hapten-specific antibody/interleukin-2 receptor chimera.
IL-2 is a cytokine that is essential for the expansion and survival of activated T cells. Although adoptive transfer of tumor-specific T cells with IL-2 is one of strategies for cancer immunotherapy, it is essential to replace IL-2 that exerts severe side effects in vivo. To solve this problem, we propose to use an antibody/IL-2R chimera, which can transduce a growth signal in response to a cognate antigen. We constructed two chimeras, in which ScFv of anti-fluorescein antibody was tethered to extracellular D2 domain of erythropoietin receptor and transmembrane/cytoplasmic domains of IL-2Rbeta or gamma chain. When the chimeras were co-expressed in IL-3-dependent pro-B cell line Ba/F3 and IL-2-dependent T cell line CTLL-2, gene-modified cells were selectively expanded in the absence of IL-3 and IL-2, respectively, by adding fluorescein-conjugated BSA (BSA-FL) as a cognate antigen. Growth assay revealed that the cells with the chimeras transduced a growth signal in a BSA-FL dose-dependent manner. Furthermore, STAT3, STAT5, ERK1/2 and Akt, which are hallmarks for IL-2R signaling, were all activated by the chimeras in CTLL-2 transfectant. We also demonstrated that the chimeras were functional in murine primary T cells. These results demonstrate that the antibody/IL-2R chimeras could substantially mimic the wild-type IL-2R and could specifically expand gene-modified T cells in the presence of the cognate antigen.
Sogo
Takahiro
T
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Kawahara
Masahiro
M
Ueda
Hiroshi
H
Otsu
Makoto
M
Onodera
Masafumi
M
Nakauchi
Hiromitsu
H
Nagamune
Teruyuki
T
D000818:Animals; D002460:Cell Line; D019281:Dimerization; D005434:Flow Cytometry; D006241:Haptens / Q000737:chemistry*; D007377:Interleukin-3 / Q000378:metabolism; D008024:Ligands; D051379:Mice; D008954:Models, Biological; D017434:Protein Structure, Tertiary; D015375:Receptors, Interleukin-2 / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012190:Retroviridae / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000378:metabolism*
D006241:Haptens; D007377:Interleukin-3; D008024:Ligands; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins