SELECTIVE DESTRUCTION OF LEUKOCYTES BY FREEZING AS A POTENTIAL MEANS OF MODULATING TISSUE IMMUNOGENICITY - MEMBRANE INTEGRITY OF LYMPHOCYTES AND MACROPHAGES
SELECTIVE DESTRUCTION OF LEUKOCYTES BY FREEZING AS A POTENTIAL MEANS OF MODULATING TISSUE IMMUNOGENICITY - MEMBRANE INTEGRITY OF LYMPHOCYTES AND MACROPHAGES
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DOI：
10.1016/0011-2240(87)90011-3
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发表时间：
1987-04-01
期刊：
影响因子：
2.7
通讯作者：
BENTON, MJ
中科院分区：
文献类型：
--
作者：
TAYLOR, MJ;BANK, HL;BENTON, MJ
现在已知，当移植组织同种异体移植物时，仅抗原识别不足以激活宿主中的淋巴细胞。目前，供体组织中的“过客”白细胞（抗原呈递细胞）被认为是主要的免疫原性刺激。使用各种方法去除这些污染的白细胞，已经能够降低同种异体移植物的免疫原性，从而提高组织同种异体移植物的存活率。这项初步研究探讨了利用冷冻生物学方法调节组织免疫原性的可能性，这是由于不同细胞类型对冷冻损伤具有公认的差异敏感性。该研究是由以下事实所推动的：在使用相对快的冷却速率进行冷冻保存后，胰岛可以分泌胰岛素以响应分级葡萄糖挑战，这对于白细胞存活来说是次优的。将大鼠外周血淋巴细胞或腹膜血淋巴细胞或腹膜渗出细胞（巨噬细胞）的批次冷却至0.3、1.5、20、75或200 ℃。C/min，使用三种不同的冷冻保存方案，据报道，产生有活力的胰岛。根据冷冻后回收的细胞数量以及评估细胞膜完整性的荧光活力测定来评价细胞存活。淋巴细胞和巨噬细胞在冷冻和解冻后的最佳存活率在0.3 - 5 ℃的冷却速率范围内。这些淋巴样细胞中有相当数量（10-40%）在20 ℃冷冻下存活。并且仅在以大于75 ℃/min的速率冷却之后。C/min是存活率降低到可忽略的水平。
It is now known, when a tissue allograft is transplanted, that antigen recognition alone is not sufficient for lymphocyte activation in the host. "Passenger" leucocytes (antigen-presenting cells) present in the donor tissue are now recognized as a major immunogenic stimulus. Removal of these contaminating leucocytes, using a variety of procedures, has enabled the immunogenicity of allografts to be reduced, thus enhancing the survival of tissue allografts. This initial study explores the possibility of using a cryobiological approach to modulating the immunogenicity of tissues by virtue of the well-recognized differential susceptibility of different cell types to freezing injury. The investigation was prompted by demonstrations that pancreatic islets can secrete insulin in response to a graded glucose challenge after cryopreservation using relatively fast cooling rates which would be expected to be suboptimal for leucocyte survival. Batches of rat peripheral blood lymphocytes, or peritoneal blood lymphocytes, or peritoneal exudate cells (macrophages) were cooled at 0.3, 1.5, 20, 75, or 200.degree. C/min using three different cryopreservation protocols reported to yield viable pancreatic islets. Cell survival was evaluated in terms of the numbers of cells recovered after freezing as well as a fluorometric viability assay which assessed the membrane integrity of cells. Optimum survival of both lymphocytes and macrophages after freezing and thawing was found at cooling rates in the range of 0.3 to 5.degree. C/min. A significant number (10-40%) of these lymphoid cells survived freezing at 20.degree. C/min and only after cooling at rates greater than 75.degree. C/min was survival reduced to a negligible level.