AAHS ASPN ASRM - 2013 Annual Meeting - American Association ...

ASRM Concurrent Scientific Paper Presentations C-2
A New Concept of Cell-Based Immunotherapy with Chimeric Cells for Acceptance of Composite Tissue Allograft
Transplants
Institution where the work was prepared: The Cleveland Clinic Foundation, Cleveland, OH, USA
Maria Siemionow, MD, PhD; Aleksandra Klimczak; Yalcin Kulahci; Galip Agaoglu; Anna Jankowska; The Cleveland Clinic Foundation
Purpose:
In search for strategies replacing life-long immunosuppression cellular therapeutics become a new alternative for solid organ transplants. This study was
designed to evaluate the efficacy of adoptive transfer of donor-specific chimeric cells, originating from two different MHC mismatched donors in engraftment
and immunotherapeutic effect of composite tissue transplants acceptance.
Methods:
Thirty three trimeras (11 primary and 22 secondary) were created across MHC barrier. First, primary trimeras (n=9) were created via intraosseous bone marrow
transplantation of 70x106 cells from LBN(RT1n) and ACI(RT1a) donors to the same LEW(RT1l) recipients. Next, secondary trimeras (n=22) were created via
adoptive transfer of MACS-sorted: double positive RT1n/RT1a (9x106-24x106), and single positive RT1n (4.5x106-9.0x106) and RT1a (6.0x106-12.0x106) cells
from primary trimeras into the bone of naïve recipients. Eight secondary trimeras served as controls without treatment, 14 trimeras received 7-day protocol of
·‚-TCR/CsA therapy. Efficacy of MACS-sorting and donor chimerism (for MHC class I antigens) was assessed by flow cytometry. At day 21 after secondary trimera
creation, the therapeutic effect of adoptive transfer was tested by bilateral transplantation of skin allografts from the LBN and ACI donors. Immunofluorescence
assessed the presence of donor cells in the lymphoid organs of recipients.
Results:
In primary trimeras 13.1% of LBN donor positive cells (RT1n) and 6.8% of ACI donor positive cell (RT1a) was found. MACS-sorting revealed 87% - 96% purity
for double positive RT1n/RT1a cells. At day 21 secondary trimeras created via transplantation of double positive cell (RT1n/RT1a) revealed 8.3% of RT1n
and 11.3% of RT1a positive cells. Transfer of single positive cells induced chimerism of 6.0% for RT1n and 7.6% for RT1a Non treatment trimeras rejected
transplanted flaps within 11 to 18 days. Prolonged skin flap survival was achieved up to 120 days after transplantation of double positive RT1n/RT1a cells and
for over 200 days in recipients receiving single positive RT1n and RT1a chimeric cells (still under observation). Presence of chimeric cells in the spleen, lymph
nodes and thymus of recipients confirmed engraftment of cells from two different donors into lymphoid organs of recipients.
Conclusions:
Intraosseous transplantation of bone marrow cells from two different MHC mismatched donors resulted in creation of donor-specific trimera. Isolation and
adoptive transfer of chimeric cells proved to be efficacious in engraftment and chimerism induction in naïve donors leading to extension of vascularized skin
allograft survival. This new strategy of cell-based immunotherapy may have direct application in clinical transfer for solid organ and CTA transplants.
Intrajejunal Administration of Fresh Donor Splenocytes Significantly Delays the Onset of Rejection of
Heterotopic Hindlimb Composite Tissue Allotransplants in Rats
Institution where the work was prepared: Department of Plastic Surgery, Chang Gung Memorial Hospital, Taipei, Taiwan
Christopher Glenn Wallace, MB, ChB, MRCS; Chia-Hung Yen, PhD; Hsiang-Chen Yang, MSc; Chun-Yen Lin, MD, PhD; Ren-Chin Wu, MD; Wei-Chao Huang, MD, PhD; Fu-
Chan Wei, MD, FACS; Chang Gung Memorial Hospital, Chang Gung University and Medical College
Antigen-specific oral/mucosal tolerance has benefited allograft survival in several animal models; jejunal mucosal tolerance was recently shown to be superiorly tolerogenic for rat cardiac
allotransplants. Therefore, it was investigated whether intrajejunal (IJ) administration of fresh donor splenocytes (FDS) could delay semi-allogeneic hindlimb CTA rejection.
Methodology:
Adult (8-10 weeks) age-/sex-matched recipient Lewis (LEW; RT1l) and donor Lewis-Brown-Norway (LBN; RT1l+n) rats were used. Five recipient Groups were
investigated: “SHAM” (n=12), “TREATED” (n=12), “SHAM/CTA” (n=5), “TREATED/CTA” (n=8) and “ISO” (isogeneic hindlimb transplants; n=5). Percutaneous
gastro-duodeno-jejunostomies were sited in all rats in the first four Groups on Day -12 via midline laparotomies. SHAM and TREATED rats were sacrificed on
Day 0 for in vitro mixed lymphocyte reaction (MLR). SHAM/CTA and TREATED/CTA instead received heterotopic LBN hindlimb CTAs on Day 0. TREATED and
SHAM received IJ-FDS (5x107 cells) or vehicle alone, respectively, everyday on Days -9 through -3. TREATED/CTA and SHAM/CTA received IJ-FDS (5x107 cells)
or vehicle alone, respectively, everyday from Day -9 until completion of rejection. Immunosuppressive drugs were never administered. CTAs were monitored for
onset and completion of rejection (using published definitions). Recipient CTA-muscle and CTA-skin were biopsied on Day +7.
Results:
In vitro MLR (Figure): Proliferation of SHAM Group splenocytes against LBN stimulation was significantly greater than that of TREATED splenocytes against
LBN stimulation (p0.05). Transplant survival: Onset
of rejection was significantly delayed by IJ-FDS administration (p