title: IGF-I triple helix gene therapy of rat and human gliomas.
abstract: OBJECTIVE
  IGF-I anti-gene technology was applied in treatment of rat and human gliomas using IGF-I triple helix approach.
  
  
  METHODS
  CNS-1 rat glioma cell and primary human glioblastoma cell lines established from surgically removed glioblastomas multiforme were transfected in vitro with IGF-I antisense (pMT-Anti-IGF-I) or IGF-I triple helix (pMT-AG-TH) expression vectors. The transfected cells were examined for immunogenicity (immunocytochemistry and flow cytometry analysis) and apoptosis phenomena (electron microscopy). 3 x 10(6) transfected cells were inoculated subcutaneously either into transgenic Lewis rats or in patients with glioblastoma. The peripheral blood lymphocytes (PBL) derived from "vaccinated" patients were immunophenotyped for the set of CD antigens (CD4, CD8 etc).
  
  
  RESULTS
  Using immunocytochemistry and Northern blot techniques, the transfected "antisense" and "triple-helix" cells showed total inhibition of IGF. Transfected cultures were positively stained either for both MHC-I and B7 antigens--60% of cloned lines, or for MHC-I only--40% of cloned lines. Moreover "triple helix" cells as compared to "antisense" cells showed slightly higher expression of MHC-I or B7. Transfected cells also showed the feature of apoptosis in 60%-70% of cells. In in vivo experiments with rats bearing tumors, the injection of "triple helix" cells expressing both MHC-I and B7 interrupted tumor growth in 80% of cases. In contrast, transfected cells expressing only MHC-I stopped development in 30% of tumors. In five patients with surgically resected glioblastoma who were inoculated with "triple helix" cells, PBL showed an increased percentage of CD4 + CD25+ and CD8 + CD11b-cells, following two vaccinations.
  
  
  CONCLUSIONS
  The anti-tumor effectiveness of IGF-I anti-gene technology may be related to both MHC-I and B7 expression in cells used for therapy. The IGF-I antigene therapy of human glioblastoma multiforme increases immune response of treated patients.
authors.affiliation: School of Medicine, Case Western Reserve University, Cleveland, USA.
authors.forename: L A
authors.initials: LA
authors.lastname: Trojan
authors.forename: P
authors.initials: P
authors.lastname: Kopinski
authors.forename: A
authors.initials: A
authors.lastname: Mazurek
authors.forename: L
authors.initials: L
authors.lastname: Chyczewski
authors.forename: A
authors.initials: A
authors.lastname: Ly
authors.forename: P
authors.initials: P
authors.lastname: Jarocki
authors.forename: J
authors.initials: J
authors.lastname: Nikliński
authors.forename: A
authors.initials: A
authors.lastname: Shevelev
authors.forename: R
authors.initials: R
authors.lastname: Trzos
authors.forename: Y
authors.initials: Y
authors.lastname: Pan
authors.forename: D J
authors.initials: DJ
authors.lastname: Gitis
authors.forename: M
authors.initials: M
authors.lastname: Bierwagen
authors.forename: J L
authors.initials: JL
authors.lastname: Czapiewska
authors.forename: M X
authors.initials: MX
authors.lastname: Wei
authors.forename: J
authors.initials: J
authors.lastname: Michalkiewicz
authors.forename: D
authors.initials: D
authors.lastname: Henin
authors.forename: T
authors.initials: T
authors.lastname: Popiela
authors.forename: F
authors.initials: F
authors.lastname: Evrard
authors.forename: H
authors.initials: H
authors.lastname: Kasprzak
authors.forename: D
authors.initials: D
authors.lastname: Anthony
authors.forename: J
authors.initials: J
authors.lastname: Trojan
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D018122:B7-1 Antigen / Q000235:genetics; D019496:Cancer Vaccines; D045744:Cell Line, Tumor; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D005909:Glioblastoma / Q000235:genetics / Q000628:therapy; D005910:Glioma / Q000235:genetics* / Q000628:therapy*; D015395:Histocompatibility Antigens Class I / Q000235:genetics; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D008297:Male; D023421:Models, Animal; D016372:RNA, Antisense; D051381:Rats; D014162:Transfection
chemical_list: D018122:B7-1 Antigen; D019496:Cancer Vaccines; D015395:Histocompatibility Antigens Class I; D016372:RNA, Antisense; D007334:Insulin-Like Growth Factor I