背景:炎症反应和凝血障碍是成功的心脏异种移植进入临床的相关挑战。为了解决这个问题,需要有效且临床可行的抗炎和抗凝血方案.通过器官来源猪的基因工程可以减少炎症和凝血反应。此外,有几种治疗策略可以预防或减少异种移植后的炎症反应和凝血障碍.然而,目前还不清楚,哪种药物组合应用于临床。为了阐明这一点,我们提供了使用几种抗炎药的组合进行的猪-狒狒原位异种心脏移植实验的数据。
方法:转基因仔猪(GGTA1-KO,hCD46/hTBM转基因)用于原位心脏异种移植到圈养的狒狒中(n=14)。所有动物都接受了包括C1酯酶抑制剂在内的抗炎药治疗,IL-6受体拮抗剂,TNF-α抑制剂,和IL-1受体拮抗剂。作为一种添加剂药物,给予乙酰水杨酸和普通肝素。免疫抑制方案基于CD40/CD40L共刺激阻断。在实验过程中,白细胞计数,在多个时间点评估C反应蛋白(CRP)水平以及全身细胞因子和趋化因子水平和凝血参数.由于猪巨细胞病毒/猪玫瑰病毒(PCMV/PRV)感染(n=2)或技术故障(n=2),将四只动物排除在进一步的数据分析之外。
结果:白细胞计数显示围手术期相关减少,CRP水平升高。在术后期间,白细胞计数始终保持在正常范围内,CRP水平在术后约35、50和80天后又显示三个峰值。细胞因子和趋化因子的分析揭示了不同的模式。一些细胞因子,像IL-8一样,在围手术期增加约2倍,但随后下降到与术前值相当的水平甚至更低。其他细胞因子,如IL-12/IL-23,在围手术期下降并保持在这些水平。除了围手术期减少,没有观察到凝血参数的相关改变.总之,所有参数均显示心脏异种移植后炎症反应和凝血功能紊乱的病程无明显变化,因此显示了我们方法的有效性.
结论:我们在抗炎药治疗方面的临床前经验证明,在病原体传播的情况下,控制异种移植中的炎症和凝血障碍是可能的,并且是可行的。尤其是PCMV/PRV对受体的影响被阻止,因为PCMV/PRV也诱导炎症和凝血障碍。我们的抗炎方案在心脏异种移植的临床环境中也应适用且有效。
BACKGROUND: Inflammatory responses and coagulation disorders are a relevant challenge for successful cardiac xenotransplantation on its way to the clinic. To cope with this, an effective and clinically practicable anti-inflammatory and anti-coagulatory regimen is needed. The inflammatory and coagulatory response can be reduced by genetic engineering of the organ-source pigs. Furthermore, there are several therapeutic strategies to prevent or reduce inflammatory responses and coagulation disorders following xenotransplantation. However, it is still unclear, which combination of drugs should be used in the clinical setting. To elucidate this, we present data from pig-to-baboon orthotopic cardiac xenotransplantation experiments using a combination of several anti-inflammatory drugs.
METHODS: Genetically modified piglets (GGTA1-KO, hCD46/hTBM transgenic) were used for orthotopic cardiac xenotransplantation into captive-bred baboons (n = 14). All animals received an anti-inflammatory drug therapy including a C1 esterase inhibitor, an IL-6 receptor antagonist, a TNF-α inhibitor, and an IL-1 receptor antagonist. As an additive medication, acetylsalicylic acid and unfractionated heparin were administered. The immunosuppressive regimen was based on CD40/CD40L co-stimulation blockade. During the experiments, leukocyte counts, levels of C-reactive protein (CRP) as well as systemic cytokine and chemokine levels and coagulation parameters were assessed at multiple timepoints. Four animals were excluded from further data analyses due to porcine cytomegalovirus/porcine roseolovirus (PCMV/PRV) infections (n = 2) or technical failures (n = 2).
RESULTS: Leukocyte counts showed a relevant perioperative decrease, CRP levels an increase. In the postoperative period, leukocyte counts remained consistently within normal ranges, CRP levels showed three further peaks after about 35, 50, and 80 postoperative days. Analyses of cytokines and chemokines revealed different patterns. Some cytokines, like IL-8, increased about 2-fold in the perioperative period, but then decreased to levels comparable to the preoperative values or even lower. Other cytokines, such as IL-12/IL-23, decreased in the perioperative period and stayed at these levels. Besides perioperative decreases, there were no relevant alterations observed in coagulation parameters. In summary, all parameters showed an unremarkable course with regard to inflammatory responses and coagulation disorders following cardiac xenotransplantation and thus showed the effectiveness of our approach.
CONCLUSIONS: Our preclinical experience with the anti-inflammatory drug therapy proved that controlling of inflammation and coagulation disorders in xenotransplantation is possible and well-practicable under the condition that transmission of pathogens, especially of PCMV/PRV to the recipient is prevented because PCMV/PRV also induces inflammation and coagulation disorders. Our anti-inflammatory regimen should also be applicable and effective in the clinical setting of cardiac xenotransplantation.