Floor plan - 2013 Annual Meeting - American Association for Hand ...

In Vivo and In Vitro Evidence that Intrajejunal Administration of Fresh Donor Splenocytes
Delays the Onset of Rejection of Hindlimb Composite Tissue Allotransplants by Regulating
Th1/Th2 Cytokines in Rats
Institution where the work was prepared: Chang Gung Memorial Hospital and Chang Gung University, Taipei, Taiwan
Christopher Glenn Wallace, MB, ChB, MRCS1; Chia-Hung Yen, PhD2; Hsiang-Chen Yang, MSc1; Chun-Yen Lin, MD,
PhD1; Wei-Chao Huang1; Jeng-Yee Lin1; Fu-Chan Wei, MD, FACS1; (1)Chang Gung Memorial Hospital, Chang Gung
University and Medical College, (2)National Pingtung University of Science and Technology
INTRODUCTION:
Jejunal mucosal tolerance was superior to oral tolerance in generating donor-specific hyporesponsiveness to cardiac allotransplants in
rats. We previously demonstrated that intrajejunal administration of fresh donor splenocytes (FDS) delayed onset of rejection of semiallogeneic
hindlimb composite tissue allotransplants (CTA) in rats, even without concurrent immunosuppressive drug therapy. We now
report our investigations into the underlying mechanism.
METHODOLOGY:
Adult (8-10 weeks) male recipient Lewis (RT1l) and donor Lewis-Brown-Norway (LBN; RT1l+n) rats were used in this study. Percutaneous
gastro-duodeno-jejunostomies were sited in all Lewis rats on Day -12. Lewis rats received either intrajejunal-FDS from LBN donors
(5x107 cells; TREATED Group; n=12) or vehicle alone (HBSS only; SHAM Group; n=11) everyday on Days -9 through -3 (7 doses) and
were sacrificed on Day 0 for one-way mixed lymphocyte reaction (MLR). Other Lewis rats received heterotopic LBN hindlimb CTAs on
Day 0, and received intrajejunal-FDS from LBN donors (5x107 cells; TREATED/CTA Group; n=8) or vehicle alone (HBSS only; SHAM/CTA
Group; n=5) everyday from Day -9 until completion of CTA rejection. Immunosuppressive drugs were never administered. Cytokine levels
were determined by ELISA (TGF-beta) or flow cytometric bead arrays (IL4, IL10, IFN-Gamma, TNF-alpha) in one-way MLR supernatants
(obtained after 72 hours co-culture) and in sera obtained from CTA recipients on Day +7 post-transplant.
RESULTS:
IL10 (p