Local overexpression of nerve growth factor in rat corneal transplants improves allograft survival.
OBJECTIVE
To investigate the effect and mechanisms of nerve growth factor (NGF) gene therapy to promote allograft survival in experimental rat corneal transplantation.


METHODS
A rat major histocompatibility complex (MHC) class I/II disparate corneal transplant model was used. Recipients were randomly assigned to receive either local Ad (Ad)-mediated gene transfer of NGF or a single intraperitoneal injection of AdNGF 1 day before transplantation. Moreover, immunosuppressive therapy was introduced by systemic coapplication of an Ad expressing CTLA4Ig. The efficacy of this treatment was examined by intracorneal mRNA expression analysis of cytokines and cytoprotective molecules by quantitative RT-PCR at day 12 after transplant. Further graft integrity and immune response against adenoviral vectors were investigated.


RESULTS
Local AdNGF-gene transfer significantly prolonged the mean survival time (MST) of rat corneal grafts (16.8 +/- 1.4 days) compared with control grafts (MST, 13.1 +/- 0.3 days; P < 0.03). In contrast, systemic AdNGF gene transfer did not result in improved corneal graft survival (MST, 15.2 +/- 1.0 days). RT-PCR analysis of cornea explants revealed diminished expression of proinflammatory cytokines (IFN-gamma, TNF-alpha) and increased expression of antiapoptotic molecules. In addition, graft endothelial integrity was improved, as measured by the detection of apoptotic cells. Moreover, coapplication of CTLA4Ig further significantly improved graft survival and protective effects of local NGF gene therapy.


CONCLUSIONS
This is the first report showing the successful application of a neurotrophin gene therapy to prolong corneal graft survival in an experimental rat transplantation model. Moreover, immunomodulatory therapy further improves graft survival and demonstrates that both anti-inflammatory and cytoprotective mechanisms are involved in the prevention of corneal allograft rejection.
Gong
Nianqiao
N
Department of Ophthalmology, Charité-University Medicine, Berlin, Germany.
Pleyer
Uwe
U
Vogt
Katrin
K
Anegon
Ignacio
I
Flügel
Alexander
A
Volk
Hans-Dieter
HD
Ritter
Thomas
T
D000069594:Abatacept; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D016039:Corneal Transplantation; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006084:Graft Rejection / Q000517:prevention & control; D006085:Graft Survival / Q000502:physiology*; D006801:Humans; D018796:Immunoconjugates / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000627:therapeutic use; D020932:Nerve Growth Factor / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011917:Rats, Inbred Lew; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014184:Transplantation, Homologous
D016207:Cytokines; D018796:Immunoconjugates; D007166:Immunosuppressive Agents; D012333:RNA, Messenger; D000069594:Abatacept; D020932:Nerve Growth Factor