Castro Grace, Merkel Patricia A, Giclas Hannah E, Gibula Andrew, Andersen Gillian E, Corash Laurence M, Lin Jin Sying, Green Jennifer, Knight Vijaya, Stassinopoulos Adonis
Cerus Corporation, Concord, California.
National Jewish Health, Denver, Colorado.
Transfusion. 2018 Jun;58(6):1506-1515. doi: 10.1111/trf.14589. Epub 2018 Apr 2.
Transfusion-associated graft-versus-host disease (TA-GVHD) is a rare complication after transfusion of components containing viable donor T cells. Gamma irradiation with doses that stop T-cell proliferation is the predominant method to prevent TA-GVHD. Treatment with pathogen inactivation methodologies has been found to also be effective against proliferating white blood cells, including T cells. In this study, T-cell inactivation was compared, between amotosalen/ultraviolet A (UVA) treatment and gamma-irradiation (2500 cGy), using a sensitive limiting dilution assay (LDA) with an enhanced dynamic range.
Matched plasma units (N = 8), contaminated with 1 × 10 peripheral blood mononuclear cells (PBMCs) per mL, were either treated with amotosalen/UVA or gamma irradiation, or retained as untreated control. Posttreatment, cells were cultured under standardized conditions. T-cell proliferation was determined by the incorporation of H-thymidine and correlated with microscopic detection.
Range-finding experiments showed that after gamma irradiation (2500 cGy), significant T-cell proliferation could be observed at a 1 × 10 cell culture density, some proliferation at 1 × 10 , and none at 1 × 10 cells/well. Based on these facts, a quantitative comparison was carried out between amotosalen/UVA at the highest challenge of 1 × 10 PBMCs/well, and gamma irradiation at 1 × 10 and 1 × 10 PBMCs/well. Complete inactivation of the T cells after amotosalen/UVA treatment was observed, equivalent to greater than 6.2 log inactivation. Complete inactivation of the T cells was also observed after gamma irradiation when 1 × 10 PBMCs/well were cultured (>4.2 log inactivation). Proliferation was observed when 1 × 10 PBMCs/well were cultured (≤5.2 log inactivation) after gamma irradiation.
Amotosalen/UVA treatment more effectively inactivates T cells than the current standard of gamma irradiation (2500 cGy) for the prevention of TA-GVHD.
输血相关移植物抗宿主病（TA-GVHD）是输注含有存活供体T细胞的成分血后的一种罕见并发症。采用能阻止T细胞增殖的剂量进行伽马射线辐照是预防TA-GVHD的主要方法。已发现采用病原体灭活方法进行处理对包括T细胞在内的增殖白细胞也有效。在本研究中，使用具有增强动态范围的灵敏极限稀释分析（LDA），比较了在阿莫沙生/紫外线A（UVA）处理与伽马射线辐照（2500 cGy）之间的T细胞灭活情况。
将每毫升污染1×10外周血单个核细胞（PBMC）的匹配血浆单位（N = 8），分别用阿莫沙生/UVA或伽马射线辐照处理，或留作未处理对照。处理后，细胞在标准化条件下培养。通过掺入³H-胸腺嘧啶核苷来测定T细胞增殖，并与显微镜检测结果相关联。
预实验表明，在伽马射线辐照（2500 cGy）后，在细胞培养密度为1×10时可观察到显著的T细胞增殖，在1×10时出现一些增殖，而在1×10细胞/孔时无增殖。基于这些事实，在每孔1×10 PBMC的最高挑战剂量下对阿莫沙生/UVA与每孔1×10和1×10 PBMC的伽马射线辐照进行了定量比较。观察到阿莫沙生/UVA处理后T细胞完全失活，相当于大于6.2对数失活。当每孔培养1×10 PBMC时，伽马射线辐照后也观察到T细胞完全失活（>4.2对数失活）。伽马射线辐照后每孔培养1×10 PBMC时观察到增殖（≤5.2对数失活）。
对于预防TA-GVHD，阿莫沙生/UVA处理比当前的伽马射线辐照标准（2500 cGy）能更有效地使T细胞失活。
