title: An IL-12/Shh-C domain fusion protein-based IL-12 autocrine loop for             sustained natural killer cell activation.
abstract: The dependency of activated natural killer (NK) cells on the continuous             support of exogenous interleukin (IL)-2 for their in vivo survival, tumor localization             and consequently, their antitumor effect, is a major obstacle for NK cell-mediated             tumor therapy. In the present study, a fusion gene between IL-12 and mouse sonic             hedgehog C-terminal domain (Shh-C) was constructed. The fusion protein was autocatalytically             processed to form cholesterol-modified IL-12 molecules and an autocrine loop of             IL-12 was established for the sustained activation of NK cells. The transduced             NK cells matured more rapidly in vitro with the enhanced expression of granule-related             proteins. NKIL-12/Shh-C cells reached the same proliferation rate as NK cells             transduced with enhanced green fluorescent protein (EGFP)/Shh-C (NKEGFP/Shh-C)             with <10-fold IL-2 supplementation, suggesting that the fusion protein reduced             the dependency of NK cells on IL-2. The amount of interferon‑γ (IFN-γ) in the             supernatants of NKIL-12/Shh-C cells 5 and 7 days after transduction was significantly             higher than that in the supernatants of NKIL-12 cells. Immunofluorescent staining             of lung tissues from B16-bearing mice which had received an intravenous injection             of lentivirus-transduced NK cells without exogenous IL-2 confirmed that donor             NK cells successfully infiltrated into the lung tissues. The survival time of             the mice which had received NKIL-12/Shh-C cells was significantly prolonged compared             to the mice which had received NKEGFP/Shh-C cells.
authors.affiliation: Department of Biological Sciences, Clemson University, Clemson,                     SC 29634, USA.
authors.forename: Lining
authors.initials: L
authors.lastname: Zhu
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Zhao
authors.forename: Yanzhang
authors.initials: Y
authors.lastname: Wei
authors.forename: William
authors.initials: W
authors.lastname: Marcotte
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Wagner
authors.forename: Xianzhong
authors.initials: X
authors.lastname: Yu
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D019898:Autocrine Communication; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D016207:Cytokines / Q000097:blood / Q000378:metabolism; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic; D005260:Female; D053823:Hedgehog Proteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007167:Immunotherapy; D018664:Interleukin-12 / Q000096:biosynthesis / Q000235:genetics*; D007376:Interleukin-2 / Q000502:physiology; D007694:Killer Cells, Natural / Q000276:immunology / Q000502:physiology* / Q000637:transplantation; D008168:Lung / Q000473:pathology; D008213:Lymphocyte Activation; D008546:Melanoma, Experimental / Q000097:blood / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D009368:Neoplasm Transplantation; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D016207:Cytokines; D053823:Hedgehog Proteins; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; C465570:Shh protein, mouse; D018664:Interleukin-12