Immunsuppressive Wirkung von Mitomycin-C-behandelten mononukleären Zellen des peripheren Blutes (MICs) in der Vaskularisierten Composite Allotransplantation

Zusammenfassung

Hintergrund Vaskularisierte Composite Allotransplantationen (VCA) ermöglichen die Wiederherstellung komplexer Gewebedefekte. Die ersten erfolgreichen allogenen Hand- und Gesichtstransplantationen haben die Forschung zur Verbesserung der immunsuppressiven Therapien stetig vorangetrieben. Die Inkubation mononukleärer Zellen des peripheren Blutes (PBMCs) mit Mitomycin C (MMC) generiert immunmodulatorisch wirksame Zellen (MICs). In vorherigen Studien konnten wir eine signifikante immunsuppressive Wirkung durch die Applikation von Donor-MICs am Tag der Transplantation zeigen. Ziel dieser Studie ist es, den optimalen Zeitpunkt der Behandlung mit MICs in der VCA zu eruieren.

Material und Methoden 60 allogene Hinterlauftransplantationen wurden in 6 experimentellen Gruppen durchgeführt. Lewis-Ratten (LEW) dienten als Spender-, Brown-Norway-Ratten (BN) als Empfängertiere. Tieren der Gruppe A wurden einmalig Spender-MICs 7 Tage präoperativ systemisch verabreicht. Gruppe B-F dienten als Kontrollgruppen. Tiere der Gruppe B erhielten keine immunsuppressive Therapie. In Gruppe C wurden unbehandelte Spender-PBMCs 7 Tage präoperativ verabreicht. Tiere der Gruppe D erhielten nur das Zellkulturmedium. Tieren der Gruppe E wurde eine Standardimmunsuppression mit Tacrolimus und Prednisolon verabreicht. In Gruppe F wurden syngene Hinterlauftransplantationen (BN→BN) durchgeführt. Der Abstoßungszeitpunkt wurde sowohl anhand klinischer Beobachtungen als auch aufgrund histologischer Parameter bestimmt.

Ergebnisse In Versuchsgruppe A zeigte sich im Vergleich zu den Kontrollgruppen B, C und D (5,5 ± 0,7, 5,3 ± 0,7 und 5,7 ± 0,5) eine signifikant früher eintretende Abstoßungsreaktion der Hinterläufe nach 3,5 ± 0,2 Tagen (p < 0,01). In den Kontrollgruppen E und F zeigte sich keine Abstoßungsreaktion.

Schlussfolgerung Die Ergebnisse der vorliegenden Studie zeigen, dass die immunmodulatorische Wirkung von MICs unmittelbar vom Applikationszeitpunkt abhängt. Nachdem in vorherigen Experimenten die Applikation von MICs am Transplantationstag eine signifikante immunsuppressive Wirkung aufwies, konnte im Rahmen dieser Studie gezeigt werden, dass die präoperative Gabe von MICs zu einer beschleunigten Abstoßung führt und damit das Überleben des Transplantates signifikant verkürzt wird. Folgestudien sind notwendig, um sowohl die Modifikation des Applikationszeitpunktes als auch die Dosis-Effekt-Beziehungen und Zellcharakteristika dieser potentiell immunsuppressiven Zellen weiter zu untersuchen.

Abstract

Background Vascularized Composite Allotransplantation (VCA) enables the restoration of complex tissue defects. Since the first successful hand and face transplants were performed, clinical and experimental research has consistently improved immunosuppressive therapies. The incubation of peripheral blood mononuclear cells (PBMCs) with mitomycin C (MMC) results in immunomodulatory cells (MICs). In previous studies, the systemic application of MICs on the day of allogeneic hind limb transplantation led to a significant immunosuppression in rats. The aim of this study is to further investigate the optimal point in time of MIC application in a complex VCA model.

Material and Methods In six groups, 60 allogeneic hind limb transplantations were performed. Fully mismatched rats were used as hind limb donors [Lewis (LEW)] and recipients [Brown-Norway (BN)]. Group A received donor-derived MICs seven days preoperatively. Group B received no immunosuppression; group C received untreated PBMCs seven days prior to transplantation. Animals in group D received cell culture media, whereas group E was treated with a standard immunosuppression consisting of tacrolimus and prednisolone. In group F, syngeneic hind limb transplantations (BN→BN) were performed. Transplant rejection was assessed clinically and histologically.

Results Group A showed a significantly earlier onset of allograft rejection after 3.5 ± 0.2 days (p < 0.01) when compared with control groups B, C and D (5.5 ± 0.7, 5.3 ± 0.7 und 5.7 ± 0.5). Groups E and F showedno allograft rejection.

Conclusion This study shows that the time of application determines the immunomodulatory effects of MICs. Whereas the systemic application of MICs on the day of transplantation led to a significant immunosuppression in previous studies, this study demonstrates that preoperative injections of MICs lead to an acceleration of allotransplant rejection. Follow-up studies are necessary to investigate further modifications of application time as well as dose-effect relations and cell characteristics of these potential immunosuppressive cells.

Schlüsselwörter

Transplantation experimentell - experimentelle Chirurgie

Key words

Vascularized composite allotransplantation - composite tissue allotransplantation - PBMCs - MICs - mitomycin C-incubated cells

Publication History

Article published online:
07 January 2021

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