Ingraft chimerism in lung transplantation--a study in a porcine model of obliterative bronchiolitis

Abstract

Background: Bronchial epithelium is a target of the alloimmune response in lung transplantation, and intact epithelium may protect allografts from rejection and obliterative bronchiolitis (OB). Herein we study the influence of chimerism on bronchial epithelium and OB development in pigs.

Methods: A total of 54 immunosuppressed and unimmunosuppressed bronchial allografts were serially obtained 2-90 days after transplantation. Histology (H&E) was assessed and the fluorescence in situ hybridization (FISH) method for Y chromosomes using pig-specific DNA-label was used to detect recipient derived cells in graft epithelium and bronchial wall, and donor cell migration to recipient organs. Ingraft chimerism was studied by using male recipients with female donors, whereas donor cell migration to recipient organs was studied using female recipients with male donors.

Results: Early appearance of recipient-derived cells in the airway epithelium appeared predictive of epithelial destruction (R=0.610-0.671 and p<0.05) and of obliteration of the bronchial lumen (R=0.698 and p<0.01). All allografts with preserved epithelium showed epithelial chimerism throughout the follow-up. Antirejection medication did not prevent, but delayed the appearance of Y chromosome positive cells in the epithelium (p<0.05), or bronchial wall (p<0.05).

Conclusions: In this study we demonstrate that early appearance of Y chromosomes in the airway epithelium predicts features characteristic of OB. Chimerism occurred in all allografts, including those without features of OB. Therefore we suggest that ingraft chimerism may be a mechanism involved in the repair of alloimmune-mediated tissue injury after transplantation.

Figure 1

Destruction of airway epithelium (A) and obliteration of the bronchial lumen (B), and the positive staining for Y chromosomes in the epithelium (C) and in the bronchial wall (D). The study groups were non-immunosuppressed, inadequately immunosuppressed, or adequately immunosuppressed. Epithelial destruction, luminal obliteration, and positive staining for Y chromosomes were graded on a scale from 0-3. The number of assessed bronchi in each group and on each assessment point was 6.6. ± 1.1 for histological samples and 7.1. ± 1.1 for FISH.

Figure 2

Table 2. Histological alterations (A,B,C) and chimerism (D,E,F) in bronchial allografts (A,B,D,E) and in a control autograft (C,F). Epithelial damage is observed in a bronchial allograft on follow-up day 4 (A) and recovered epithelium and bronchial wall structure on day 7 in an adequately treated allograft (B). H&E, original magnification ×10. Recovery of the adequately treated allograft (B) was similar to that of a control autograft (C) on day 7. H&E, original magnification ×10. Tissue section of a bronchial allograft of female origin in a male recipient hybridized with probes for pig chromosomes Y (red) and 1 (green) on day 7 (D) shows epithelial and bronchial wall cells with Y chromosomes. Y chromosomes are more frequent on follow-up day 10 (E) than on follow-up day 7 (D) in recovered epithelium in adequately treated allografts. Chromosomes Y (red) and 1 (green) stain invariably (F) in a male control autograft on follow-up day 7 and confirm successful hybridization. Staining with 4',6-diamidino-2-phenylindole, original magnification ×1000.