[CTLA4-FasL protein for the prevention of immune rejection in mouse corneal transplantation].
OBJECTIVE
To investigate the therapeutic effect of double-functioned CTLA4-FasL protein for the prevention of immune rejection in murine corneal allografts.


METHODS
It was a experimental study. C57BL/6 mice (n = 45) were as donors and BALB/c mice (n = 90) as recipients. BALB/c mice accepted penetrating keratoplasty were randomly divided into 3 groups including no therapy (A), CsA DDS implanted in anterior chamber (B), 10 microg/mL CTLA4-FasL (C). Survival time of corneal allografts was observed and routine assays were performed including immunohistochemistry, reverse-transcription polymerase chain reaction (RT-PCR), TdT-mediated dUTP nick end labeling (TUNEL).


RESULTS
Survival time of corneal transplants was (14.3 +/- 1.3) days, (58.0 +/- 2.8) days, (106.3 +/- 17.5) days respectively. There was significant difference between groups in statistics analysis (P = 0.000). Inflammatory cells, predominant for CD4 + T cells, showed an increasing tendency for rejected corneal allografts in A group, peaked on postoperative 7 d and then decreased in C group compared with invisible in B group. CD80 or CD86 was detected on postoperative 3 d or 7 d respectively in excised cornea and iris of both A and B group, while either showed weakening expression in C group. On postoperative 14 d, IL-10, TNF-alpha, IFN-gamma mRNA were only detected in rejected cornea allograft of A group other than clear allografts of B or C group. Compared with no apoptosis in the other groups, abundant apoptosis cells were visible in cornea and iris of C group on postoperative 7 d.


CONCLUSIONS
CTLA4-FasL is able to prolong survival time of corneal allografts by dual action mechanism to exert immunosuppressive effects, both blockade of CD28-CD80/86 pathway and induction of apoptosis of T cell.
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Qingdao 266071, China.
Min
M
Chen
Wei-yun
WY
Shi
Fu-hua
FH
Wang
Li-xin
LX
Xie
D000818:Animals; D015703:Antigens, CD / Q000276:immunology / Q000627:therapeutic use*; D017209:Apoptosis; D018122:B7-1 Antigen / Q000276:immunology; D051940:B7-2 Antigen / Q000276:immunology; D018106:CD28 Antigens / Q000276:immunology; D060908:CTLA-4 Antigen; D016207:Cytokines / Q000276:immunology; D053222:Fas Ligand Protein / Q000276:immunology / Q000627:therapeutic use*; D006084:Graft Rejection / Q000517:prevention & control*; D015948:Keratoplasty, Penetrating; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013601:T-Lymphocytes / Q000276:immunology; D014184:Transplantation, Homologous
D015703:Antigens, CD; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D018106:CD28 Antigens; D060908:CTLA-4 Antigen; C556707:Ctla4 protein, mouse; D016207:Cytokines; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse