Donor Recipient Chimeric Cells Induce Chimerism and Extend Survival of Vascularized Composite Allografts

Arkadiusz Jundzill,Maria Siemionow,Aleksandra Klimczak,Maria Madajka-Niemeyer,Joanna Cwykiel

doi:10.1007/s00005-021-00614-9

Abstract

This study evaluated the efficacy of donor recipient chimeric cell (DRCC) therapy created by fusion of donor and recipient derived bone marrow cells (BMC) in chimerism and tolerance induction in a rat vascularized composite allograft (VCA) model. Twenty-four VCA (groin flaps) from MHC-mismatched ACI (RT1a) donors were transplanted to Lewis (RT1l) recipients. Rats were randomly divided into (n = 6/group): Group 1—untreated controls, Groups 2—7-day immunosuppression controls, Group 3—DRCC, and Group 4—DRCC with 7-day anti-αβTCR monoclonal antibody and cyclosporine A protocol. DRCC created by polyethylene glycol-mediated fusion of ACI and Lewis BMC were cultured and transplanted (2–4 × 106) to VCA recipients via intraosseous delivery route. Flow cytometry assessed peripheral blood chimerism while fluorescent microscopy and PCR tested the presence of DRCC in the recipient’s blood, bone marrow (BM), and lymphoid organs at the study endpoint (VCA rejection). No complications were observed after DRCC intraosseous delivery. Group 4 presented the longest average VCA survival (79.3 ± 30.9 days) followed by Group 2 (53.3 ± 13.6 days), Group 3 (18 ± 7.5 days), and Group 1 (8.5 ± 1 days). The highest chimerism level was detected in Group 4 (57.9 ± 6.2%) at day 7 post-transplant. The chimerism declined at day 21 post-transplant and remained at 10% level during the entire follow-up period. Single dose of DRCC therapy induced long-term multilineage chimerism and extended VCA survival. DRCC introduces a novel concept of customized donor-recipient cell-based therapy supporting solid organ and VCA transplants.

Highlights

The continued effort to introduce new solutions into the field of transplantation is driven by the need to limit the use of anti-rejection protocols which, necessary, are associated with severe co-morbidities and significant shortening of transplant recipient’s lifespan.Cell-based therapies were proposed as a promising supportive treatment due to observed direct and indirect involvement of cells of hematopoietic and mesenchymal origin in the alteration of the local and systemic immune response of the transplant recipients (Siemionow et al 2012)
The average vascularized composite allotransplantation (VCA) survival time ranged from 8–125 days post-transplant (PT) and showed the following average survival times: Group 1 8.5 ± 1 days PT, Group 2 (VCA recipients under 7-day anti-αβTCR/CsA IS protocol) 53.3 ± 13.6 days PT (p < 0.001 vs. Group 1), Group 3 (VCA recipients supported with donor-recipient chimeric cells (DRCC) therapy) 18 ± 7.5 days PT (p = 0.01 vs. Group 1 and p < 0.001 vs. Group 2), and Group 4 (VCA recipients supported with DRCC therapy and 7-day antiαβTCR/CsA IS protocol) 79.3 ± 30.9 days PT
The VCA survivals in Group 4 were significantly prolonged in comparison to untreated VCA recipient controls (Group 1, p < 0.001), VCA recipients group supported with 7-day IS protocol (Group 2, p < 0.05) and VCA recipients supported with DRCC therapy (Group 3, p < 0.001)

Summary

Introduction

The continued effort to introduce new solutions into the field of transplantation is driven by the need to limit the use of anti-rejection protocols which, necessary, are associated with severe co-morbidities and significant shortening of transplant recipient’s lifespan.Cell-based therapies were proposed as a promising supportive treatment due to observed direct and indirect involvement of cells of hematopoietic and mesenchymal origin in the alteration of the local and systemic immune response of the transplant recipients (Siemionow et al 2012). Archivum Immunologiae et Therapiae Experimentalis (2021) 69:13 the occurrence of cells presenting phenotype and/or genotype make-up specific for both the transplant donor and recipient and suggested contribution of the donor/recipient cells to the processes such as regeneration and immune response (Alvarez-Dolado et al 2003; Camargo et al 2004; Johansson et al 2008; LaBarge and Blau 2002; Lluis and Cosma 2010; Nygren et al 2004; Powell et al 2011; Rizvi et al 2006; Sanges et al 2013; Vassilopoulos et al 2003; Wang et al 2003; Weimann et al 2003; Willenbring et al 2004). The improvement of VCA survival time could have been caused by the combination of multi-lineage chimerism induction and/or immunomodulatory properties of in vivo created DRCC (Hivelin et al 2016)

Methods

Results

Conclusion