Laboratory of Biochemistry and Vascular Biology, Division of Blood Components and Devices, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States of America.
Laboratory of Cellular Hematology, Division of Blood Components and Devices, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States of America.
Blood Transfus. 2024 Jul 15;22(4):316-327. doi: 10.2450/BloodTransfus.718.
Quality assessment of modified or processed red blood cell (RBC) components, such as pathogen-reduced RBCs, using only in vitro testing may not always be predictive of in vivo performance. Mouse or rat in vivo models are limited by a lack of applicability to certain aspects of human RBC biology. Here, we used a guinea pig model to study the effects of riboflavin combined with UV light on the integrity of RBCs in vitro and following transfusion in vivo.
Guinea pig RBCs were collected from whole blood (WB) treated with varying UV doses (10, 20, 40 or 80 J/mL) in the presence of riboflavin (UVR-RBCs). In vitro tests for UVR-RBCs included hemolysis, osmotic fragility, and cellular morphology by scanning electron microscopy. Guinea pigs transfused with one-day post-treatment UVR-RBCs were evaluated for plasma hemoglobin (Hb), non-transferrin bound iron (NTBI), total iron and Perls-detectable hemosiderin deposition in the spleen and kidney, and renal uptake of Hb.
Acute RBC injury was dose dependently accelerated after treatment with UV light in the presence of riboflavin. Aberrant RBC morphology was evident at 20, 40, and 80 J/mL, and membrane lysis with Hb release was prominent at 80 J/mL. Guinea pigs transfused with 40 and 80 J/mL UVR-RBCs showed increased plasma Hb levels, and plasma NTBI was elevated in all UVR-RBC groups (10-80 J/mL). Total iron levels and Perls-hemosiderin staining in spleen and kidney as well as Hb uptake in renal proximal tubules were increased 8 hours post-transfusion with 40 and 80 J/mL UVR-RBCs.
UVR-RBCs administered to guinea pigs increased markers of intravascular and extravascular hemolysis in a UV dose-dependent manner. This model may allow for the discrimination of RBC injury during testing of extensively processed RBCs intended for transfusion.
使用仅在体外进行的测试来评估改良或加工的红细胞（RBC）成分的质量，例如病原体减少的 RBC，可能并不总是能预测体内的性能。鼠或大鼠的体内模型由于缺乏对人类 RBC 生物学某些方面的适用性而受到限制。在这里，我们使用豚鼠模型来研究核黄素与紫外线联合作用对 RBC 体外完整性和体内输血后完整性的影响。
从用不同紫外线剂量（10、20、40 或 80 J/mL）处理的全血（WB）中收集豚鼠 RBC，并在核黄素存在下进行紫外线照射（UVR-RBCs）。用于 UVR-RBC 的体外测试包括溶血、渗透脆性和扫描电子显微镜下的细胞形态。对输注一天后 UVR-RBC 的豚鼠进行血浆血红蛋白（Hb）、非转铁蛋白结合铁（NTBI）、总铁和 Perls 可检测铁在脾脏和肾脏中的沉积以及 Hb 在肾脏中的摄取的评估。
在核黄素存在下用紫外线照射后，RBC 急性损伤呈剂量依赖性加速。在 20、40 和 80 J/mL 时可见异常 RBC 形态，在 80 J/mL 时可见膜裂解和 Hb 释放。输注 40 和 80 J/mL UVR-RBC 的豚鼠显示血浆 Hb 水平升高，所有 UVR-RBC 组（10-80 J/mL）的血浆 NTBI 均升高。输注 40 和 80 J/mL UVR-RBC 后 8 小时，脾脏和肾脏中的总铁水平和 Perls-血铁黄素染色以及肾近端小管中的 Hb 摄取均增加。
给豚鼠输注 UVR-RBC 会以紫外线剂量依赖的方式增加血管内和血管外溶血的标志物。这种模型可能允许在对用于输血的经过广泛处理的 RBC 进行测试时区分 RBC 损伤。
