Abstract
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 immunologically 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, survive after grafting and induce host sensitization to alloantigen. Methods: CEAs from CBA donors (H- 2(k)) were grown on allogeneic NIH 3T3 (H-2(q)) or syngeneic LTK (H-2(k)) fibroblasts, which were removed by trypsinization 7 days later. CBA mice (n = 85) were flank-grafted with NIH allografts (positive control), CEA/3T3s, CEA/LTKs, or CBA autografts (negative control). Hosts were challenged with second set NIH tail allografts 3 weeks later. Median graft survival was compared between groups by Wilcoxon rank and λ2 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 immunoblotting, 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 determined by alloantigen expression on Western immunoblots. Accelerated tail 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. Conclusions: Immunogenic fibroblasts used to grow CEAs survive in vivo and sensitize the graft recipient for accelerated second-set rejection. These persistent cells may initiate an inflammatory response that may result in late graft breakdown and limit the utility of CEAs grown with a foreign fibroblast feeder layer.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 51-60 |
| Number of pages | 10 |
| Journal | Journal of Trauma - Injury, Infection and Critical Care |
| Volume | 41 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jul 1 1996 |
| Externally published | Yes |
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Keywords
- Burn injury
- Cultured epidermal autograft
- Skin replacement
ASJC Scopus subject areas
- Surgery
- Critical Care and Intensive Care Medicine
Cite this
Allogeneic fibroblasts used to grow cultured epidermal autografts persist in vivo and sensitize the graft recipient for accelerated second-set rejection. / Hultman, C. Scott; Brinson, George M.; Siltharm, Soranit; DeSerres, Suzan; Cairns, Brauce A.; Peterson, H. D.; Meyer, Anthony A.
In: Journal of Trauma - Injury, Infection and Critical Care, Vol. 41, No. 1, 01.07.1996, p. 51-60.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Allogeneic fibroblasts used to grow cultured epidermal autografts persist in vivo and sensitize the graft recipient for accelerated second-set rejection
AU - Hultman, C. Scott
AU - Brinson, George M.
AU - Siltharm, Soranit
AU - DeSerres, Suzan
AU - Cairns, Brauce A.
AU - Peterson, H. D.
AU - Meyer, Anthony A.
PY - 1996/7/1
Y1 - 1996/7/1
N2 - 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 immunologically 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, survive after grafting and induce host sensitization to alloantigen. Methods: CEAs from CBA donors (H- 2(k)) were grown on allogeneic NIH 3T3 (H-2(q)) or syngeneic LTK (H-2(k)) fibroblasts, which were removed by trypsinization 7 days later. CBA mice (n = 85) were flank-grafted with NIH allografts (positive control), CEA/3T3s, CEA/LTKs, or CBA autografts (negative control). Hosts were challenged with second set NIH tail allografts 3 weeks later. Median graft survival was compared between groups by Wilcoxon rank and λ2 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 immunoblotting, 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 determined by alloantigen expression on Western immunoblots. Accelerated tail 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. Conclusions: Immunogenic fibroblasts used to grow CEAs survive in vivo and sensitize the graft recipient for accelerated second-set rejection. These persistent cells may initiate an inflammatory response that may result in late graft breakdown and limit the utility of CEAs grown with a foreign fibroblast feeder layer.
AB - 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 immunologically 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, survive after grafting and induce host sensitization to alloantigen. Methods: CEAs from CBA donors (H- 2(k)) were grown on allogeneic NIH 3T3 (H-2(q)) or syngeneic LTK (H-2(k)) fibroblasts, which were removed by trypsinization 7 days later. CBA mice (n = 85) were flank-grafted with NIH allografts (positive control), CEA/3T3s, CEA/LTKs, or CBA autografts (negative control). Hosts were challenged with second set NIH tail allografts 3 weeks later. Median graft survival was compared between groups by Wilcoxon rank and λ2 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 immunoblotting, 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 determined by alloantigen expression on Western immunoblots. Accelerated tail 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. Conclusions: Immunogenic fibroblasts used to grow CEAs survive in vivo and sensitize the graft recipient for accelerated second-set rejection. These persistent cells may initiate an inflammatory response that may result in late graft breakdown and limit the utility of CEAs grown with a foreign fibroblast feeder layer.
KW - Burn injury
KW - Cultured epidermal autograft
KW - Skin replacement
UR - http://www.scopus.com/inward/record.url?scp=0029939524&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029939524&partnerID=8YFLogxK
U2 - 10.1097/00005373-199607000-00009
DO - 10.1097/00005373-199607000-00009
M3 - Article
C2 - 8676424
AN - SCOPUS:0029939524
VL - 41
SP - 51
EP - 60
JO - Journal of Trauma and Acute Care Surgery
JF - Journal of Trauma and Acute Care Surgery
SN - 2163-0755
IS - 1
ER -