Beneficial Effects of Inhalation of Carbon Monoxide on Pulmonary Allograft Survival in MHC-inbred CLAWN Miniature Swine.

一氧化碳（CO）作为血红素催化的副产物内源性产生，并已被证明在小鼠模型中多种器官中提供对缺血-再灌注损伤（IRI）的保护。由于一氧化碳也是一种已知的有毒气体，因此在将其用于临床之前，必须使用大型动物进行转化研究。为了将CO应用于临床，我们研究了(1)在mhc近交系clwn微型猪中，供体和受体围手术期吸入CO是否能延长移植物存活时间(2)仅供体治疗就足以延长移植物存活时间，以及(2)术后CO治疗是否有额外的生存益处。18头claw猪接受了完全MHC错配的肺，并给予他克莫司12天（0-11天，35-45 ng/ml）。第1组（n=6）仅给予他克莫司。在第二组（n=5）中，供体和受体均吸入200-250 ppm CO（供体180分钟，受体390分钟，直到2小时再灌注）。在第三组（n=4）中，只有供体接受CO治疗180分钟。在第4组（n=3）中，术后14天，接受者每天额外给予1小时250 ppm CO。第1组所有受者在术后第63天出现细胞毒性抗供体抗体，一致排斥移植物。相比之下，2组5名受者中有4名接受移植超过63天。仅用供体（组3）短期CO治疗可有效延长同种异体移植物的存活时间。四分之三的受者接受移植超过63天，最长存活时间为150天。在第4组，3名受者中有2名维持移植物91天；然而，与第二组相比，术后CO并没有延长移植物的存活时间。在接受CO治疗的动物中，MLR和CML测定的抗供者抗体和供者特异性反应性的发展延迟。此外，在POD2活检中，炎症细胞浸润和内皮细胞损伤较少，血清促炎细胞因子（IL-1β, IL-6）浓度也降低。这些数据表明，在临床相关的大型动物模型中，仅供体一氧化碳治疗足以提高同种异体肺移植的存活率。在器官采集或保存过程中，用CO进行供体细胞保护预处理可以在临床环境中完成，潜在地提高移植物存活率，对受体的不良影响风险很小。少

Carbon monoxide (CO) is produced endogenously as a byproduct of heme catalysis, and has been shown to provide protection against ischemia-reperfusion injury (IRI) in a variety of organs in murine models. Since CO is also known to be a toxic gas, translational research using large animals is essential prior to its use in the clinic. To attempt CO to apply for the clinical setting, we examined whether (1) perioperative CO inhalation to both donor and recipient prolongs lung graft survival (2) donor treatment alone is sufficient to prolong graft survival and (2) postoperative CO therapy has additional survival benefits in MHC-inbred CLAWN miniature swine.Eighteen CLAWN swine received fully MHC mismatched lungs with 12 days of tacrolimus (days 0-11 ; 35-45 ng/ml). In Group 1 (n=6), recipients received tacrolimus alone. In Group 2 (n=5), both donors and recipients inhaled 200-250 ppm CO (180 min for donors ; 390 min for recipients until 2-hr reperfusion). In Group 3 (n=4), only donors were … More treated with CO for 180 min. In Group 4 (n=3), recipients were additionally given daily 1-hour 250 ppm CO for 14 postoperative days.All recipients in Group 1 uniformly rejected their grafts by postoperative day 63 with development of cytotoxic antidonor antibodies. In contrast, 4 out of 5 recipients in Group 2 accepted their grafts beyond 63 days. Short-term CO treatment only with donor (Group 3) was effective in prolonging allograft survival. Three out of 4 recipients accepted their grafts beyond 63 days with the longest survivor lasting 150 days. In Group 4, two out of 3 recipients maintained their grafts for 91 days ; however, postoperative CO did not prolong graft survival compared to Group 2. Development of antidonor-antibodies and donor-specific responsiveness by MLR and CML assays was delayed in animals that received CO therapy. Furthermore, fewer inflammatory cell infiltrates and injuries of endothelial cells were seen on POD2 biopsies, and serum concentrations of the pro-inflammatory cytokines (IL-1β, IL-6) also decreased in Group 2-4.These data suggest that donor CO treatment alone is sufficient to improve lung allograft survival in a clinically relevant large animal model. Donor cytoprotective preconditioning with CO during organ harvest or preservation could be accomplished in a clinical setting, potentially improving graft survival with little risk of adverse effects on the recipient. Less