Mathes David W, Solari Mario G, Randolph Mark A, Gazelle G Scott, Yamada Kazuhiko, Huang Christene A, Sachs David H, Lee W P Andrew
Division of Plastic Surgery, Harvard Medical School, Massachusetts General Hospital, Boston, MA, USA.
Transplantation. 2005 Nov 15;80(9):1300-8. doi: 10.1097/01.tp.0000178933.31987.11.
The aim was to investigate if intravascular in utero injection of adult bone marrow into swine fetuses could lead to macrochimerism and tolerance to the donor.
Outbred Yorkshire sows and boars screening negative for MHC allele SLA of MGH miniature swine were bred. A laparotomy was performed on the sows at 50 days gestation to expose the uterus. Bone marrow harvested from SLA miniature swine was T-cell depleted and injected intravascularly into seventeen fetuses. Flow cytometry was performed to detect donor cells (chimerism) in the peripheral blood after birth. Mixed lymphocyte reactions (MLR) and cell-mediated lympholysis (CML) assays were used to assess the response to donor MHC. Previously frozen skin grafts from the bone marrow donor were placed on the offspring from the first litter. Donor-matched renal transplant from SLA donors were performed on chimeric swine, with and without a short 12-day course of cyclosporine, and one nonchimeric littermate.
Nine inoculated offspring demonstrated donor cell chimerism in the peripheral blood and lymphohematopoietic tissues. All animals with detectable chimerism within the first three weeks were consistently nonreactive to donor MHC in vitro. Animals challenged with donor skin grafts displayed prolonged graft survival without producing antidonor antibodies. All chimeric animals accepted donor-matched kidney allografts, even one without cyclosporine. The kidney in the nonchimeric littermate rejected by day 21.
Transplantation of allogeneic adult bone marrow into immunocompetent fetal recipients resulted in chimerism. In utero inoculation led to operational tolerance to the donor's major histocompatibility antigens and long-term acceptance to organ allografts.
目的是研究在子宫内将成年骨髓血管内注射到猪胎儿体内是否会导致大嵌合体形成以及对供体的耐受性。
选择对MGH小型猪的MHC等位基因SLA筛查呈阴性的远交约克夏母猪和公猪进行繁殖。在妊娠50天时对母猪进行剖腹手术以暴露子宫。从SLA小型猪采集的骨髓进行T细胞清除，然后血管内注射到17只胎儿体内。出生后进行流式细胞术检测外周血中的供体细胞（嵌合体）。采用混合淋巴细胞反应（MLR）和细胞介导的淋巴细胞溶解（CML）试验评估对供体MHC的反应。将先前冷冻的骨髓供体皮肤移植到第一窝仔猪身上。对嵌合猪以及一只非嵌合同窝仔猪进行来自SLA供体的供体匹配肾移植，其中部分猪接受为期12天的环孢素短期治疗。
9只接种的后代在外周血和淋巴造血组织中表现出供体细胞嵌合现象。在前三周内所有检测到嵌合现象的动物在体外对供体MHC始终无反应。接受供体皮肤移植的动物移植存活期延长，且未产生抗供体抗体。所有嵌合动物均接受了供体匹配的肾移植，甚至有一只未接受环孢素治疗的也成功接受了移植。非嵌合同窝仔猪的肾脏在第21天时发生排斥反应。
将同种异体成年骨髓移植到具有免疫活性的胎儿受体中可导致嵌合体形成。子宫内接种可导致对供体主要组织相容性抗原的操作性耐受以及对器官同种异体移植的长期接受。
