Cs. Hultman et al., ALLOGENEIC FIBROBLASTS USED TO GROW CULTURED EPIDERMAL AUTOGRAFTS PERSIST IN-VIVO AND SENSITIZE THE GRAFT RECIPIENT FOR ACCELERATED 2ND-SETREJECTION, The journal of trauma, injury, infection, and critical care, 41(1), 1996, pp. 51-58
Introduction: Cultured epidermal autografts (CEAs) have been used for
wound coverage in patients with massive burns and other skin defects,
However, CEAs often display late breakdown, which may be immunological
ly mediated and initiated by persistent foreign fibroblasts used as a
feeder layer to optimize keratinocyte growth, This study investigates
whether these fibroblasts, previously shown to persist in vitro, survi
ve after grafting and induce host sensitization to alloantigen. Method
s: CEAs from CBA donors (H-2(k)) were grown ore allogeneic NIH 3T3 (H-
2(q)) or syngeneic LTK (H-2(k)) fibroblasts, which were removed by try
psinization 7 days later, CBA mice (n = 85) were flank-grafted with NI
H allografts (positive control), CEA/3T3s, CEA/LTKs, or CBA autografts
(negative control). Hosts were challenged with second set NIH tail al
lografts 3 weeks later, Median graft survival was compared between gro
ups by Wilcoxon rank and chi(2) analysis, Additional CBA mice (n = 15)
received CEAs that were biopsied 0, 4, and 8 days after grafting. The
presence of allogeneic fibroblasts was determined by Western immunobl
otting, using KL295, a monoclonal antibody that recognizes H-2(q) (but
not H-2(k)) class II histocompatibility antigens. Results: Allogeneic
fibroblasts persisted after grafting but decreased over time, as dete
rmined by alloantigen expression on Western immunoblots. Accelerated t
ail graft rejection occurred in hosts primed by NIH allografts (9 days
, p < 0.05), as well as by CEAs grown with an allogeneic (10 days, p <
0.05) but not a syngeneic feeder layer (12 days, NS), Mice receiving
flank autografts rejected second set tail allografts at 12 days. Concl
usions: Immunogenic fibroblasts used to grow CEAs survive in vivo and
sensitize the graft recipient for accelerated second-set rejection. Th
ese persistent cells may initiate an inflammatory response that may re
sult in late graft breakdown and limit the utility of CEAs grown with
a foreign fibroblast feeder layer.