Antigen-mediated growth control of hybridoma cells via a human artificial chromosome.
Human artificial chromosome (HAC) vectors possess several characteristics sufficient for the requirements of gene therapy vectors, including stable episomal maintenance and mediation of long-term transgene expression. In this study, we adopted an antigen-mediated genetically modified cell amplification (AMEGA) system employing an antibody/cytokine receptor chimera that triggers a growth signal in response to a cognate non-toxic antigen, and applied it to growth control of HAC-transferred cells by adding an antigen that differed from cytokines that may manifest pleiotropic effects. We previously constructed a novel HAC vector, 21 Delta qHAC, derived from human chromosome 21, housed in CHO cells. Here, we constructed an HAC vector harboring an ScFv-gp130 chimera responsive to fluorescein-conjugated BSA (BSA-FL) as well as a model transgene, enhanced green fluorescent protein (EGFP), in CHO cells. The modified HAC was transferred into interleukin (IL)-6-dependent hybridoma 7TD1 cells by microcell-mediated chromosome transfer, and the cells were subsequently found to show BSA-FL-dependent cell growth and sustained expression of EGFP in the absence of IL-6. The AMEGA system in combination with HAC technology will be useful for increasing the efficacy of gene therapy by conferring a growth advantage on the genetically modified cells.
Kawahara
Masahiro
M
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. kawahara@bio.t.u-tokyo.ac.jp
Inoue
Toshiaki
T
Ren
Xianying
X
Sogo
Takahiro
T
Yamada
Hidetoshi
H
Katoh
Motonobu
M
Ueda
Hiroshi
H
Oshimura
Mitsuo
M
Nagamune
Teruyuki
T
D000818:Animals; D016466:CHO Cells; D002455:Cell Division / Q000502:physiology*; D002460:Cell Line; D022222:Chromosomes, Artificial, Human / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D006825:Hybridomas / Q000166:cytology*; D015850:Interleukin-6 / Q000494:pharmacology; D051379:Mice; D011993:Recombinant Fusion Proteins; D014162:Transfection
D015850:Interleukin-6; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins