The fleur-de-lis upper gracilis flap for breast reconstruction: flap design and outcome.

We evaluated a fleur-de-lis design for the gracilis myocutaneous flap to improve flap volume for breast reconstruction. Thirty-one flaps were used in 17 consecutive patients undergoing the procedure for either thin body habitus (23 flaps) or prior abdominal surgery (8 flaps). The flap success rate was 100%. The fleur-de-lis flap provided proportionate breast reconstructions in all patients. Complications included 6 (19.3%) donor-site dehiscence and 4 (12.9%) episodes of cellulitis. Applying a negative pressure dressing to the donor site (n=26) significantly reduced the initially high dehiscence rate to 7.7% (P<0.01). There was no incidence of lower extremity edema or sensory loss. The fleur-de-lis gracilis flap can be performed with a low flap related complication rate and acceptable donor-site morbidity. Because of its standardized flap design, improved volume, and favorable breast shaping, it may allow autologous breast reconstruction to be offered to a greater number of patients.

Human immune responses to porcine endogenous retrovirus-derived peptides presented naturally in the context of porcine and human major histocompatibility complex class I molecules: implications in xenotransplantation of porcine organs.

BACKGROUND: Porcine endogenous retroviruses (PERV) have been shown to infect human cells, raising concerns regarding safety of xenotransplantation. In patients exposed to porcine tissues, no PERV infection has been observed. This study was designed to develop human CD8+ cytotoxic T lymphocytes (CTL) against PERV-derived peptides presented in the context of human leukocyte antigen (HLA) or swine leukocyte antigen (SLA) class I molecules and to define dominant epitopes contributed by PERV. METHODS: Human CD8+ CTL were generated against porcine aortic endothelial cells (PAEC). Peptides presented on SLA class I molecules were acid eluted and fractionated by reverse-phase high-performance liquid chromatography. Peptide fractions that restored lysis of acid-stripped PAEC were sequenced by tandem mass spectrometry. Human CD8+ CTL were generated against PERV envelope-derived peptides and PERV-infected human cells to identify immunodominant PERV-derived epitopes. RESULTS: We identified two peptides derived from retroviral transactivating regulatory protein (AHQDPLPEQP) and retroviral transcription factor (PQKPFVT) recognized by human CD8+ CTL in the context of SLA class I. Computer-assisted analysis identified nine PERV-envelope-derived 9-mer peptides with high affinity for the HLA-A2 molecule (Env-1-9). PERV-specific CD8+ CTL generated in vitro identified the immunodominant Env-5 peptide (303-311, KLFSLIQGA) and demonstrated HLA-A2-restricted cytotoxicity against PERV-infected human cells. CONCLUSIONS: Our results indicate that PERV-derived peptides are presented naturally on porcine and human major histocompatibility complex class I molecules. CD8+ CTL responses elicited against dominant SLA and HLA class I-restricted PERV-derived epitopes may play an important role in xenograft rejection and in containment of PERV infection of human cells after xenotransplantation.

Natural antibodies prevent in vivo transmission of porcine islet-derived endogenous retrovirus to human cells.

The discovery of porcine endogenous retroviruses (PERV) has raised concerns regarding the safety of porcine xenotransplantation. However, transmission of PERV had not been observed in humans exposed to porcine tissue. We examined whether PERV derived from porcine pancreatic islet cells could infect human cells in vivo and the role of natural antibodies in inhibiting PERV infection. In vivo infective potential of PERV was studied in SCID mice reconstituted with human peripheral blood leucocytes. Porcine islets were transplanted under the kidney capsule. PERV infection was determined by analyzing PERV gene expression in graft infiltrating lymphocytes (GIL) harvested 21 days posttransplantation. Mice were administered normal human serum prior to and 2 days posttransplantation to study their role in protection of human cells against PERV infection. PERV genes were expressed in all porcine tissues examined, including purified porcine islets. PERV expression was detected in GILs from three of five human-SCID mice. Administration of human serum blocked PERV infection in GILs in five of five human-SCID mice. These results indicate that PERV from porcine islets can infect human cells in vivo. Normal human serum blocks transmission of retrovirus in vivo, suggesting that natural xenoreactive antibodies can prevent PERV infection.

Pirfenidone inhibits obliterative airway disease in a murine heterotopic tracheal transplant model.

BACKGROUND: Chronic lung allograft rejection in the form of bronchiolitis obliterans syndrome and its histopathologic correlate, obliterative bronchiolitis (OB), are a major source of morbidity and mortality after lung transplantation. Murine heterotopic tracheal transplants into fully allogeneic mismatched recipients develop obliterative airway disease (OAD), which is a suitable model of OB. Using this murine heterotopic tracheal allograft model, we evaluated the effect of pirfenidone, a novel antifibrotic agent, on the development of OAD. METHODS: Mice transplanted with complete MHC-mismatched tracheal allografts received pirfenidone (0.5%) in pulverized food according to different schedules: daily for the first 14 days after transplantation or daily for the duration of the study beginning on posttransplantation days 0, 5, or 10. RESULTS: Mice on a continuous daily regimen of pirfenidone failed to develop evidence of chronic allograft rejection at the termination of the study (60 days). Mice receiving pirfenidone limited to the early posttransplantation period had delayed onset of OAD to 60 days. Forty percent (2/5) of mice receiving a continuous regimen of pirfenidone beginning on day 5 after transplantation had no evidence of OAD at 28 days. However, when the drug was started on day 10, all mice developed OAD by 28 days. CONCLUSIONS: Our results demonstrate a delay of onset or abrogation of OAD when pirfenidone is administered in the early posttransplantation period. These findings suggest that pirfenidone is a candidate drug to be evaluated for prevention of the fibrotic changes seen in OB in human recipients of lung transplants.

Xenoreactive anti-Galalpha(1,3)Gal antibodies prevent porcine endogenous retrovirus infection of human in vivo.

The discovery of porcine endogenous retroviruses (PERV) has raised concerns regarding the safety of pig to human xenotransplantation. In this study, we examined PERV infection of human cells in vivo. Furthermore, we examined the effect of human xenoreactive natural antibody on in vivo PERV infection. Human peripheral blood leukocyte reconstituted severe combined immunodeficiency mice were transplanted with porcine aortic endothelial cells (PAEC). PERV gene expression was readily detected in human leukocytes after transplantation. In contrast, human leukocytes harvested from mice treated with human serum or anti-Galalpha(1,3)Gal antibody prevented PERV infection in 6 of 8 mice. These results provide the first evidence that PERV can infect human cells in vivo and that natural xenoreactive antibody can prevent this infection.