title: Transplantation of transduced retinal pigment epithelium in rats.
abstract: OBJECTIVE
  To examine the effects of transplanting retinal pigment epithelial (RPE) cells transduced with neurotrophic factor genes into the subretinal space of rats.
  
  
  METHODS
  RPE cells were transduced with plasmids carrying the cDNAs of Axokine (ciliary neurotrophic factor [CNTF]; Sumitomo Pharmaceuticals Co., Ltd., Tokyo, Japan), brain derived-neurotrophic factor (BDNF), and basic fibroblast growth factor (bFGF) genes. These RPE cells were transplanted into the subretinal space of rats, and the localization was examined. The expression of enhanced green fluorescent protein (eGFP)-BDNF-transduced RPE in the subretinal space was examined by real-time polymerase chain reaction (PCR) after the transplanted cells were collected by cell sorting. The expression of major histocompatibility complex (MHC) class I and -II after gene transduction was examined by real-time PCR. The ratio of CD4(+) and CD8(+) T cells and antibody production against transplanted cells were analyzed by flow cytometry.
  
  
  RESULTS
  The transplant sites were not significantly different among the neurotrophic factors tested. The RPE cells expressed the BDNF gene in the subretinal region at approximately the same level as that in vitro. RPE cells transduced with Axokine stimulated MHC-I expression, and the cell transplantation changed the ratio of CD4(+) and CD8(+) T cells. A significant production of antibody against the Axokine-transduced RPE cells was also observed after Axokine-transduced RPE transplantation.
  
  
  CONCLUSIONS
  RPE cells transduced with neurotrophic factors express the factors after transplantation into the subretinal space. RPE transduced with Axokine or bFGF, in contrast to RPE transduced with BDNF, stimulate an immunologic reaction of the host.
authors.affiliation: Department of Ophthalmology, School of Medicine, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Saigo
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Abe
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Hojo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tomita
authors.forename: Eriko
authors.initials: E
authors.lastname: Sugano
authors.forename: Makoto
authors.initials: M
authors.lastname: Tamai
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D016516:CD4-CD8 Ratio; D017690:Cell Transplantation; D020934:Ciliary Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000378:metabolism* / Q000637:transplantation*; D010957:Plasmids; D051381:Rats; D020318:Rats, Long-Evans; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000378:metabolism / Q000601:surgery*; D014161:Transduction, Genetic
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D020934:Ciliary Neurotrophic Factor; D018076:DNA, Complementary; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D008164:Luminescent Proteins; D009414:Nerve Growth Factors; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins; C118937:axokine