Uterine rejection after allogeneic uterus transplantation in the rat is effectively suppressed by tacrolimus  Shamima N. Akhi, M.D., Cesar Diaz-Garcia, M.D., Randa R. El-Akouri, M.D., Ph.D., Caiza A. Wranning, Ph.D., Johan Mölne, M.D., Ph.D., Mats Brännström, M.D., Ph.D.  Fertility and Sterility  Volume 99, Issue 3, Pages 862-870 (March 2013) DOI: 10.1016/j.fertnstert.2012.11.002 Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Schematic diagram of the uterine graft (light gray) transplanted orthotopically with vascular end-to-side common iliac–common iliac anastomosis and vaginal end-to-end anastomosis. The venous system is indicated in dark gray; the arterial system is indicated in white. Surgical clips were used at the end of the bicornuate native uterus and at the tip of the grafted uterus, which were approximated to the native uterus by a single suture. Fertility and Sterility 2013 99, 862-870DOI: (10.1016/j.fertnstert.2012.11.002) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Light microscopy of native uteri from sham controls (A: Sham-surgery; B: Sham-TAC) and of uterine grafts of the transplanted groups (C: UTx-nonTAC; D: UTx-TAC). (A) Uterus of Sham-surgery group exhibiting normal epithelial lining (E), stroma with glands (arrow), and muscle layers (M). (B) Uterus of Sham-TAC group showing low numbers of leukocytes, mainly eosinophils, in the stroma and normal glands (arrow). The endometrium (E) and muscle layer (M) are of normal appearance. (C) Necrotic allograft (UTx-nonTAC) with central necrosis (n) and hemorrhage and only a brim of inflamed muscular tissue remaining. (D) Transplanted uterus of a tacrolimus-treated animal (UTx-TAC) with normal morphology including glands (arrow). Bar = 100 μm. Fertility and Sterility 2013 99, 862-870DOI: (10.1016/j.fertnstert.2012.11.002) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Density (panels A–C) and immunohistochemistry (panels D–F) concerning T-lymphocytes in uterine tissue sections. Density of CD4+ (A), CD8+ (B), and CD3+ (C) T-lymphocytes in the endometrium (striped bars) and myometrium (crossed bars) in the native uteri of sham groups treated with surgery (Sham-surgery) or with tacrolimus (Sham-TAC) and in the uterine graft of tacrolimus-treated animals (UTx-TAC). Dots indicate medians, bars indicate the 10%–90% range, and boxes indicate the 25%–75% range. *P<.05; n = 5–7 in each group. The arrows of panels D–F indicate CD8-positive T-cells in the native uteri of sham controls (D: Sham-surgery; E: Sham-TAC) and tacrolimus-treated transplanted uterus (F: UTx-TAC). There are very low numbers of CD8-positive T-cells in all tissue sections. An occasional CD8+ cell is seen in the epithelial layer of the tacrolimus-treated UTx-TAC group (triangle). Bar = 40 μm. Fertility and Sterility 2013 99, 862-870DOI: (10.1016/j.fertnstert.2012.11.002) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Expression levels of mRNA for IL-1α (A), IP-10 (B), and galectin-1 (C). Dots indicate medians, bars indicate the 10%–90% range, and boxes indicate the 25%–75% range. Relative mRNA levels in sham-operated non-tacrolimus-treated uterus (Sham-surgery), nontransplanted tacrolimus-treated uterus (Sham-TAC), nontreated transplanted uterus (UTx-nonTAC), and tacrolimus-treated transplanted uterus (UTx-TAC) were calculated by the ΔΔCT method and are expressed in relation to internal β-actin control. *P<.05; **P<.01. n = 5–7 in each group. Fertility and Sterility 2013 99, 862-870DOI: (10.1016/j.fertnstert.2012.11.002) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions