Mapping the Diversity of the Innate Immune System over Time After Left Lung Transplantation in Mice

Abstract

Purpose Following lung transplantation, the surgical intervention with ischemic periods and the foreign allograft are the first to trigger a response from the innate immune system, which represents the primary nonspecific defense against “non-self”. In this study, we investigated the diversity of the emerging innate immune cells and the timing of their appearance in a murine orthotopic left lung transplant model. Methods Mouse orthotopic left lung transplantation was performed in isografts (C57BL/6 to C57BL/6) and allografts (Balbc to C57BL/6). All mice received daily immunosuppression of 10 mg/kg cyclosporin A and 1.6 mg/kg methylprednisolone. Serial sacrifice was performed at day 1, 7 and 35 post-transplantation (n=6 at each time point for each group). Left transplanted lungs were harvested, single cell suspension was made and absolute numbers of innate immune cells were quantified by flow cytometry. Results Following transplantation, isografts and allografts showed an increase in neutrophil (p=0.017) and tissue resident NK cell (p=0.002) numbers at day 1, and in eosinophil (p=0.002) numbers at day 7 post-lung transplant. Granulocytes demonstrated similar patterns in isografts and allografts. Allografts demonstrated an increase in antigen presenting cells i.e. patrolling monocytes (Ly6C-) (p=0.002), interstitial macrophages (p<0.0001) and dendritic cells (p=0.005) at day 7 compared to isografts. Classical NK cells were increased in allografts at day 7 versus isografts (p=0.03). All cell numbers returned to baseline isograft values at day 35 post-transplant, except for dendritic cells (p=0.0045). Conclusion Diverse innate immune cell types show different patterns over time post-lung transplantation depending on transplant induced ischemia-reperfusion injury and/or alloimmune response against the foreign graft. These differences may be very important to dissect the various pathophysiological processes triggered by lung transplantation. Following lung transplantation, the surgical intervention with ischemic periods and the foreign allograft are the first to trigger a response from the innate immune system, which represents the primary nonspecific defense against “non-self”. In this study, we investigated the diversity of the emerging innate immune cells and the timing of their appearance in a murine orthotopic left lung transplant model. Mouse orthotopic left lung transplantation was performed in isografts (C57BL/6 to C57BL/6) and allografts (Balbc to C57BL/6). All mice received daily immunosuppression of 10 mg/kg cyclosporin A and 1.6 mg/kg methylprednisolone. Serial sacrifice was performed at day 1, 7 and 35 post-transplantation (n=6 at each time point for each group). Left transplanted lungs were harvested, single cell suspension was made and absolute numbers of innate immune cells were quantified by flow cytometry. Following transplantation, isografts and allografts showed an increase in neutrophil (p=0.017) and tissue resident NK cell (p=0.002) numbers at day 1, and in eosinophil (p=0.002) numbers at day 7 post-lung transplant. Granulocytes demonstrated similar patterns in isografts and allografts. Allografts demonstrated an increase in antigen presenting cells i.e. patrolling monocytes (Ly6C-) (p=0.002), interstitial macrophages (p<0.0001) and dendritic cells (p=0.005) at day 7 compared to isografts. Classical NK cells were increased in allografts at day 7 versus isografts (p=0.03). All cell numbers returned to baseline isograft values at day 35 post-transplant, except for dendritic cells (p=0.0045). Diverse innate immune cell types show different patterns over time post-lung transplantation depending on transplant induced ischemia-reperfusion injury and/or alloimmune response against the foreign graft. These differences may be very important to dissect the various pathophysiological processes triggered by lung transplantation.