title: Capacity of human beta-defensin expression in gene-transduced and cytokine-induced cells.
abstract: The purpose of this study was to determine the capacity of cells transduced with human beta-defensins (HBDs) to express antimicrobial peptides, since sufficient expression level is required for effective antimicrobial activity. Retroviral vector pBabeNeo and lentiviral vector SIN18cPPTRhMLV (SIN18) carrying HBDs were utilized to transduce non-HBD-expressing cells such as fibroblasts or HBD-producing oral epithelial cells. We found that HBD-3 gene transfer to fibroblasts was possible not via retrovirus but by direct vector transfection. SIN18 had high transduction efficiencies (80.9-99.9%) and transduced cells expressed higher amounts of HBD-2 than those by pBabeNeo. Primary human gingival epithelial cells (HGECs) expressed greater amounts of HBD-2 than primary fibroblasts after lentiviral transduction. Additionally, HBD-2 secretion from transduced HGECs cells was further increased when stimulated with IL-1 or TNFalpha. Our data indicate that while HBD-2 expression is limited in primary fibroblasts, its expression in HGECs may be maximized by gene transduction plus cytokine induction.
authors.affiliation: Division of Associated Clinical Specialties, Section of Endodontics, UCLA School of Dentistry, Los Angeles, CA, USA.
authors.forename: Chunyi
authors.initials: C
authors.lastname: Yin
authors.forename: Hoa N
authors.initials: HN
authors.lastname: Dang
authors.forename: Hai-Bo
authors.initials: HB
authors.lastname: Zhang
authors.forename: Farzad
authors.initials: F
authors.lastname: Gazor
authors.forename: Daniel
authors.initials: D
authors.lastname: Kim
authors.forename: Ole E
authors.initials: OE
authors.lastname: Sorensen
authors.forename: George T-J
authors.initials: GT
authors.lastname: Huang
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D004847:Epithelial Cells / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005822:Genetic Vectors; D005881:Gingiva / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D051379:Mice; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D023083:beta-Defensins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D016207:Cytokines; C106641:DEFB4A protein, human; D007375:Interleukin-1; D014409:Tumor Necrosis Factor-alpha; D023083:beta-Defensins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins