title: Immunological characterization of glioblastoma cells for immunotherapy.
abstract: The aim of this study was the immunological characterization of glioblastoma cells. Glioblastoma cell lines were cultured in serum and serum-free neurobasal (NBE) medium conditions. These cell lines were characterized by flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), western blot and natural killer (NK) cell-cytotoxicity assays. A previously described NK cell expansion method that uses K562 cells expressing interleukin (IL)-15 and 4-1 BB Ligand (BBL) (K562-mb15-41BBL) was used. RT-PCR and western blots for the expression of tumor-associated antigens (TAAs), were carried out in 32 glioblastoma and seven normal brain tissues. U87 and U343 tumor cell lines showed increased expression for major histocompatibility complex (MHC)-I and -II molecules. No significant differences in the levels of CD133, MHC class I/II, MHC class I-related chain A (MICA), MICB, UL16 binding protein 1-3 (ULBP 1-3) expression in these cell lines and in NK cell cytotoxicity were observed between serum and NBE conditions. Regardless of culture conditions, U87 and U343 cell lines were sensitive to expanded NK cells, with median cytotoxicities at 4:1 effector/target ratio of 43.2% and 46.5%, respectively. In RT-PCR, U343 and U87 showed the expression of most TAAs at a high ratio compared with U251. Western blots demonstrated positive expression for BIRC5, CD99 and ERBB2 in U251, U87 and U343 cell lines and tissues. These highly-expressed TAAs such as BIRC5, CD99 and ERBB2 in glioblastoma tissue could be the targets for immunotherapy. U87 and U343 cell lines could be useful for studying the efficacy of immunotherapy related to various TAAs and NK cell immunotherapy.
authors.affiliation: Department of Neurosurgery, Chonnam National University Hwasun Hospital, Jeollanam-do, Republic of Korea.
authors.forename: Tae-Young
authors.initials: TY
authors.lastname: Jung
authors.forename: Yoo-Duk
authors.initials: YD
authors.lastname: Choi
authors.forename: Young-Hee
authors.initials: YH
authors.lastname: Kim
authors.forename: Je-Jung
authors.initials: JJ
authors.lastname: Lee
authors.forename: Hyung-Seok
authors.initials: HS
authors.lastname: Kim
authors.forename: Ju-Sun
authors.initials: JS
authors.lastname: Kim
authors.forename: Sang-Ki
authors.initials: SK
authors.lastname: Kim
authors.forename: Shin
authors.initials: S
authors.lastname: Jung
authors.forename: Duck
authors.initials: D
authors.lastname: Cho
mesh_terms: D000071443:12E7 Antigen; D015703:Antigens, CD / Q000096:biosynthesis; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D045744:Cell Line, Tumor; D005805:Genes, MHC Class I; D005802:Genes, MHC Class II; D005909:Glioblastoma / Q000276:immunology* / Q000628:therapy*; D006801:Humans; D007167:Immunotherapy; D051034:Inhibitor of Apoptosis Proteins / Q000096:biosynthesis; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis; D000077022:Survivin
chemical_list: D000071443:12E7 Antigen; D015703:Antigens, CD; D015815:Cell Adhesion Molecules; D051034:Inhibitor of Apoptosis Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D000077022:Survivin; C107489:BIRC5 protein, human; C061828:CD99 protein, human; C508053:ERBB2 protein, human
keywords: Characterization; U251; U343 cells; U87; glioblastoma; immunotherapy; tumor-associated antigens