Gorantla Vijay S, Prabhune Kaustubha A, Perez-Abadia Gustavo, Ildstad Suzanne T, Maldonado Claudio, Orhun Haldun I, Majzoub Ramsey K, Francois Cedric G, Kakoulidis Thanos P, Brouha Pascal C R, Anderson Gary L, Pidwell Diane J, Breidenbach Warren C, Barker John H
Division of Plastic and Reconstructive Surgery, Plastic Surgery Research, University of Louisville, 511 South Floyd Street, Louisville, KY 40292, USA.
Transplantation. 2003 Apr 15;75(7):922-32. doi: 10.1097/01.TP.0000058302.45424.03.
Mixed allogeneic chimerism (MAC) has been shown to induce tolerance to composite tissue allografts (CTA). However, transplantation of unmanipulated donor-specific limbs results in severe graft-versus-host disease (GVHD). This suggests that nontolerant mature donor-derived cells in the CTA may affect the stability of chimerism, potentially resulting in GVHD. The aim of this study was to develop an approach to study and prevent GVHD in a mixed chimeric-rat hind-limb transplantation model.
[ACI-->WF] chimeras received a limb from Wistar Furth (WF) (syngeneic), Fisher (third-party), or ACI (irradiated [1,050 cGy] or nonirradiated) rats. In vitro tolerance was assessed using mixed lymphocyte reactivity (MLR) assays at the time the animals were killed.
[ACI-->WF] chimeras with greater than 85% chimerism exhibited rejection-free survival of donor-specific hind limbs. However, 100% of these animals developed lethal GVHD 22.4+/-2.8 days after limb transplantation. [ACI-->WF] chimeras that underwent transplantation with irradiated ACI or syngeneic WF limbs showed no signs of rejection or GVHD at 5 months. Nonchimeric and third-party controls rejected limbs within 10 days.
Conditioning of the host WF rats with 950 cGy of irradiation (sublethal, myeloablative) led to high levels of MAC without GVHD. The mature T-cell content of nonirradiated donor (ACI) limbs was sufficient to induce lethal GVHD in 100% of tolerant mixed chimeric [ACI-->WF] hosts. Irradiation of donor limbs before transplantation resulted in long-term donor-specific tolerance and prevented GVHD. These data demonstrate that (1) established chimeras could be susceptible to GVHD caused by immunocompetent donor cells transferred with the hind limb, and (2) inactivating these cells with irradiation prevents GVHD and destabilization of chimerism, and permits rejection-free graft acceptance.
混合异基因嵌合体（MAC）已被证明可诱导对复合组织同种异体移植物（CTA）的耐受。然而，移植未经处理的供体特异性肢体可导致严重的移植物抗宿主病（GVHD）。这表明CTA中未耐受的成熟供体来源细胞可能影响嵌合体的稳定性，从而可能导致GVHD。本研究的目的是开发一种方法，用于研究和预防混合嵌合大鼠后肢移植模型中的GVHD。
[ACI→WF]嵌合体接受来自Wistar Furth（WF）（同基因）、Fisher（第三方）或ACI（照射[1,050 cGy]或未照射）大鼠的肢体。在处死动物时，使用混合淋巴细胞反应（MLR）试验评估体外耐受性。
嵌合率大于85%的[ACI→WF]嵌合体表现出供体特异性后肢的无排斥存活。然而，这些动物中有100%在肢体移植后22.4±2.8天发生致命的GVHD。接受照射的ACI或同基因WF肢体移植的[ACI→WF]嵌合体在5个月时未出现排斥或GVHD迹象。非嵌合体和第三方对照在10天内排斥肢体。
用950 cGy照射（亚致死性、骨髓清除性）处理宿主WF大鼠可导致高水平的MAC且无GVHD。未照射的供体（ACI）肢体中的成熟T细胞含量足以在100%耐受的混合嵌合[ACI→WF]宿主中诱导致命的GVHD。移植前照射供体肢体可导致长期的供体特异性耐受并预防GVHD。这些数据表明：（1）已建立的嵌合体可能易受与后肢一起转移的具有免疫活性的供体细胞引起的GVHD影响；（2）用照射使这些细胞失活可预防GVHD和嵌合体的不稳定，并允许无排斥地接受移植物。
