Nr. 2 - 2. Quartal 2001
Methodological Investigations in the
Prevention of Skin Graft Rejection Using the CD95L (FasLigand) System
– Preliminary Step in the Engineering of Immune-Privileged Skin
Allografts
A. D. Niederbichler, O. Burghard, Th. Mühlberger, M.
Koeller, A. Kassen, C. Kuhnen, K.-M. Müller, H.-U. Steinau, P. M Vogt
Introduction: Skin grafting of the severely
burned patient may cause various problems, such as limitation of
autologous skin donor sites, infection and sepsis. The prolonged time
period necessary for cultivation of autologous epidermal cells in turn
could further aggravate already existing complications of these
patients. The development of allogeneic skin grafts with reduced
immunogenicity could be a possible solution for these problems.
CD95-ligand, a transmembrane protein of the tumor-necrosis-factor
superfamily, is known to induce apoptosis via cross-linking of
CD95-expressing activated cytotoxic t-cells. Co-transplantation of
CD95-ligand may prolong graft survival. As Sertoli cells
constitutively express the Fas–Ligand, we investigated the survival
of transplanted Sertoli cells without the presence of a stabilizing
matrix as well as using a collagen-glycosaminoglycane-matrix as a
dermal substitute in the rat model.
Methods: 24 male Lewis and DA rats (150 – 170 g) were used.
Four Lewis rats and four DA rats served as testes donors. Primary
culture of Sertoli cells was performed as described previously.
Detection of the Fas-Ligand was done by flow-cytometric analysis using
a monoclonal anti-Fas-ligand antibody. Sertoli cells were spun into a
collagen-GAG-matrix (integraä , LifeSciences Corp., Plainsboro, USA)
and subsequently analyzed histologically. The 1.5 x 1.5 cm
Sertoli-cell-containing (250.000 cells / graft) collagen-GAG-matrix
was implanted subdermally in 12 male rats on the left and right
lateral thorax as follows:
1. Lewis (donor) on Lewis (recipient): 2
2. Lewis (donor) on Davis (recipient): 4
3. Davis (donor) on Davis (recipient): 2
4. Davis (donor) on Lewis (recipient): 4
5. Conventional Davis to Lewis allogeneic skin grafts: 4
LEW rats receiving DA testicular tissue and DA skin
grafts served as controls. Biopsies were taken on post-operative days
12 and 24.
Results: Allogeneic skin grafts were rejected within 12 days
while Sertoli cells survived up to 24 days in the collagen-GAG matrix.
Immunohistochemical analysis of the biopsies has shown expression of
the Fas-ligand in the dermal Sertoli-cell-populated matrix.
Conclusions: Co-transplantation of Sertoli-cell-seeded
collagen-GAG matrix is possible. The co-transplantation of
sertoli-collagen-GAG-matrix with allogeneic skin grafts may prolong
graft survival or survival of other co-transplanted cell types by Fas
/ Fas-ligand induced T-cell apoptosis.
Key words: Allogeneic skin grafts, graft rejection, CD95 / CD95L, apoptosis, sertoli cells, collagen-glycosaminoglycane matrix (integraä )
Prof. Dr. Peter M. Vogt
Klinikum Hannover Oststadt
Klinik für Plastische Hand- und Wiederherstellungschirurgie der
Medizinische Hochschule Hannover, Podbielskistr. 380
D-30659 Hannover
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