Abstract：
Diagnosing lung injury is a challenge in lung transplantation. It has been unclear if a single biopsy specimen is truly representative of the entire organ. Our objective was to investigate lung inflammatory biomarkers using human lung tissue biopsies and ex vivo lung perfusion perfusate.
Eight human donor lungs declined for transplantation were air inflated, flash frozen, and partitioned from apex to base. Biopsies were then sampled throughout the lung. Perfusate was sampled from 4 lung lobes in 8 additional donor lungs subjected to ex vivo lung perfusion. The levels of interleukin-6, interleukin-8, interleukin-10, and interleukin-1β were measured using quantitative reverse transcription polymerase chain reaction from lung biopsies and enzyme-linked immunosorbent assay from ex vivo lung perfusion perfusate.
The median intra-biopsy equal-variance P value was .50 for messenger RNA biomarkers in tissue biopsies. The median intra-biopsy coefficient of variance was 18%. In donors with no apparent focal injuries, the biopsies in each donor showed no difference in various lung slices, with a coefficient of variance of 20%. The exception was biopsies from the lingula and injured focal areas that demonstrated larger differences. Cytokines in ex vivo lung perfusion perfusate showed minimal variation among different lobes (coefficient of variance = 4.9%).
Cytokine gene expression in lung biopsies was consistent, and the biopsy analysis reflects the whole lung, except when specimens were collected from the lingula or an area of focal injury. Ex vivo lung perfusion perfusate also provides a representative measurement of lung inflammation from the draining lobe. These results will reassure clinicians that a lung biopsy or an ex vivo lung perfusion perfusate sample can be used to inform donor lung selection.
Eight human donor lungs declined for transplantation were air inflated, flash frozen, and partitioned from apex to base. Biopsies were then sampled throughout the lung. Perfusate was sampled from 4 lung lobes in 8 additional donor lungs subjected to ex vivo lung perfusion. The levels of interleukin-6, interleukin-8, interleukin-10, and interleukin-1β were measured using quantitative reverse transcription polymerase chain reaction from lung biopsies and enzyme-linked immunosorbent assay from ex vivo lung perfusion perfusate.
The median intra-biopsy equal-variance P value was .50 for messenger RNA biomarkers in tissue biopsies. The median intra-biopsy coefficient of variance was 18%. In donors with no apparent focal injuries, the biopsies in each donor showed no difference in various lung slices, with a coefficient of variance of 20%. The exception was biopsies from the lingula and injured focal areas that demonstrated larger differences. Cytokines in ex vivo lung perfusion perfusate showed minimal variation among different lobes (coefficient of variance = 4.9%).
Cytokine gene expression in lung biopsies was consistent, and the biopsy analysis reflects the whole lung, except when specimens were collected from the lingula or an area of focal injury. Ex vivo lung perfusion perfusate also provides a representative measurement of lung inflammation from the draining lobe. These results will reassure clinicians that a lung biopsy or an ex vivo lung perfusion perfusate sample can be used to inform donor lung selection.
摘要：
目的：肺移植中诊断肺损伤是一个挑战。尚不清楚单个活检标本是否真正代表整个器官。我们的目的是使用人肺组织活检和离体肺灌注灌注液研究肺部炎症生物标志物。
方法：八个因移植而拒绝的人类供体肺被充气，闪光冷冻，从顶点到底部。然后在整个肺取样活检。从经受离体肺灌注的8个额外供体肺中的4个肺叶取样灌注液。使用来自肺活检的定量逆转录聚合酶链反应和来自离体肺灌注灌注液的酶联免疫吸附测定来测量白细胞介素6，白细胞介素8，白细胞介素10和白细胞介素1β的水平。
结果：组织活检中信使RNA生物标志物的活检内等方差P值为.50。活检内变异系数中位数为18%。在没有明显局灶性损伤的捐赠者中，每个供体的活检显示不同的肺切片没有差异，变异系数为20%。例外的是舌部和受伤的病灶区域的活检显示出较大的差异。离体肺灌注灌注液中的细胞因子在不同的叶之间显示最小的变化（变异系数=4.9％）。
结论：肺活检中细胞因子基因表达是一致的，活检分析反映了整个肺，除非标本是从舌骨或局灶性损伤区域收集的。离体肺灌注灌注液还提供了来自引流叶的肺部炎症的代表性测量。这些结果将使临床医生放心，肺活组织检查或离体肺灌注灌注液样品可用于告知供体肺选择。
方法：八个因移植而拒绝的人类供体肺被充气，闪光冷冻，从顶点到底部。然后在整个肺取样活检。从经受离体肺灌注的8个额外供体肺中的4个肺叶取样灌注液。使用来自肺活检的定量逆转录聚合酶链反应和来自离体肺灌注灌注液的酶联免疫吸附测定来测量白细胞介素6，白细胞介素8，白细胞介素10和白细胞介素1β的水平。
结果：组织活检中信使RNA生物标志物的活检内等方差P值为.50。活检内变异系数中位数为18%。在没有明显局灶性损伤的捐赠者中，每个供体的活检显示不同的肺切片没有差异，变异系数为20%。例外的是舌部和受伤的病灶区域的活检显示出较大的差异。离体肺灌注灌注液中的细胞因子在不同的叶之间显示最小的变化（变异系数=4.9％）。
结论：肺活检中细胞因子基因表达是一致的，活检分析反映了整个肺，除非标本是从舌骨或局灶性损伤区域收集的。离体肺灌注灌注液还提供了来自引流叶的肺部炎症的代表性测量。这些结果将使临床医生放心，肺活组织检查或离体肺灌注灌注液样品可用于告知供体肺选择。
