Human Immune Therapy Center, University of Virginia, Charlottesville, VA, USA.
J Transl Med. 2011 Mar 8;9:26. doi: 10.1186/1479-5876-9-26.
Clinical trials of immunologic therapies provide opportunities to study the cellular and molecular effects of those therapies and may permit identification of biomarkers of response. When the trials are performed at multiple centers, transport and storage of clinical specimens become important variables that may affect lymphocyte viability and function in blood and tissue specimens. The effect of temperature during storage and shipment of peripheral blood on subsequent processing, recovery, and function of lymphocytes is understudied and represents the focus of this study.
Peripheral blood samples (n = 285) from patients enrolled in 2 clinical trials of a melanoma vaccine were shipped from clinical centers 250 or 1100 miles to a central laboratory at the sponsoring institution. The yield of peripheral blood mononuclear cells (PBMC) collected before and after cryostorage was correlated with temperatures encountered during shipment. Also, to simulate shipping of whole blood, heparinized blood from healthy donors was collected and stored at 15 °C, 22 °C, 30 °C, or 40 °C, for varied intervals before isolation of PBMC. Specimen integrity was assessed by measures of yield, recovery, viability, and function of isolated lymphocytes. Several packaging systems were also evaluated during simulated shipping for the ability to maintain the internal temperature in adverse temperatures over time.
Blood specimen containers experienced temperatures during shipment ranging from -1 to 35 °C. Exposure to temperatures above room temperature (22 °C) resulted in greater yields of PBMC. Reduced cell recovery following cryo-preservation as well as decreased viability and immune function were observed in specimens exposed to 15 °C or 40 °C for greater than 8 hours when compared to storage at 22 °C. There was a trend toward improved preservation of blood specimen integrity stored at 30 °C prior to processing for all time points tested. Internal temperatures of blood shipping containers were maintained longer in an acceptable range when warm packs were included.
Blood packages shipped overnight by commercial carrier may encounter extreme seasonal temperatures. Therefore, considerations in the design of shipping containers should include protecting against extreme ambient temperature deviations and maintaining specimen temperature above 22 °C or preferably near 30 °C.
免疫疗法的临床试验提供了研究这些疗法的细胞和分子作用的机会，并可能识别出反应的生物标志物。当试验在多个中心进行时，临床标本的运输和储存成为重要的变量，这些变量可能会影响血液和组织标本中淋巴细胞的活力和功能。储存和运输过程中温度对随后处理、恢复和功能的影响淋巴细胞研究不足，这是本研究的重点。
从参加黑色素瘤疫苗两项临床试验的患者中采集外周血样本（n=285），由 250 英里或 1100 英里外的临床中心运往赞助机构的中央实验室。在冷冻储存前后收集的外周血单核细胞（PBMC）的产量与运输过程中遇到的温度相关。此外，为了模拟全血的运输，从健康供体采集肝素化血液，并在 15°C、22°C、30°C 或 40°C 下储存不同时间，然后分离 PBMC。通过测量分离淋巴细胞的产量、恢复、活力和功能来评估标本完整性。在模拟运输过程中，还评估了几种包装系统在一段时间内保持不利温度下内部温度的能力。
血液标本容器在运输过程中经历的温度范围为-1 至 35°C。暴露于室温以上（22°C）的温度会导致 PBMC 产量增加。与在 22°C 下储存相比，在 15°C 或 40°C 下储存超过 8 小时时，冷冻保存后细胞回收减少，活力和免疫功能降低。与所有测试时间点相比，在进行处理之前在 30°C 下储存的血液标本完整性保存趋势得到改善。当包括暖包时，血液运输容器的内部温度可以在可接受的范围内保持更长时间。
通过商业承运人 overnight 运输的血液包裹可能会遇到极端的季节性温度。因此，在设计运输容器时应考虑防止环境温度的极端偏差，并将标本温度保持在 22°C 以上，最好接近 30°C。
