University of Washington, Veterans Affairs Puget Sound Health Care System, Seattle, WA 98108, USA.
Diabetes Metab Res Rev. 2011 Nov;27(8):737-45. doi: 10.1002/dmrr.1245.
Type 1 diabetes (T1D) is a cell-mediated autoimmune disease characterized by destruction of the pancreatic islet cells. The use of cryopreserved cells is preferable to the use of freshly isolated cells to monitor clinical trials to decrease assay and laboratory variability.
The T-Cell Workshop Committee of the Immunology of Diabetes Society compared two widely accepted T-cell freezing protocols (warm and cold) to freshly isolated peripheral blood mononuclear cells from patients with T1D and controls in terms of recovery, viability, cell subset composition, and performance in functional assays currently in use in T1D-related research. Nine laboratories participated in the study with four different functional assays included.
The cold freezing method yielded higher recovery and viability compared with the warm freezing method. Irrespective of freezing protocol, B cells and CD8+ T cells were enriched, monocyte fraction decreased, and islet antigen-reactive responses were lower in frozen versus fresh cells. However, these results need to take in to account that the overall response to islet autoantigens was low in some assays.
In the current study, none of the tested T-cell functional assays performed well using frozen samples. More research is required to identify a freezing method and a T-cell functional assay that will produce responses in patients with T1D comparable to responses using fresh peripheral blood mononuclear cells.
1 型糖尿病（T1D）是一种以胰岛细胞破坏为特征的细胞介导的自身免疫性疾病。与使用新鲜分离的细胞相比，使用冷冻保存的细胞更有利于监测临床试验，以降低检测和实验室的变异性。
糖尿病免疫学会的 T 细胞研讨会委员会将两种广泛接受的 T 细胞冷冻方案（温冻和冷冻）与 T1D 患者和对照组的新鲜分离外周血单个核细胞进行了比较，从恢复、活力、细胞亚群组成和当前用于 T1D 相关研究的功能检测方面进行了比较。共有 9 个实验室参与了这项研究，其中包括 4 种不同的功能检测。
与温冻法相比，冷冻法的回收率和活力更高。无论采用何种冷冻方案，冷冻后的 B 细胞和 CD8+T 细胞均富集，单核细胞分数减少，胰岛抗原反应性降低，但需要注意的是，一些检测中的胰岛自身抗原反应总体较低。
在目前的研究中，使用冷冻样本的 T 细胞功能检测均表现不佳。需要进一步研究以确定一种冷冻方法和 T 细胞功能检测，该方法能够在 T1D 患者中产生与使用新鲜外周血单个核细胞相似的反应。
