混合淋巴细胞反应中体液同种免疫的评估。
Assessment of Humoral Alloimmunity in Mixed Lymphocyte Reaction.
作者信息
Pissas Georgios, Eleftheriadis Theodoros
机构信息
Department of Nephrology, Faculty of Medicine, University of Thessaly, Larissa, Greece.
出版信息
Bio Protoc. 2019 Jan 20;9(2):e3139. doi: 10.21769/BioProtoc.3139.
Humoral alloimmunity remains a significant and unresolved problem that constrains allograft survival. Thus, there is a need for the development of an easy, preferably non-radioactive, and inexpensive protocol for assessing the effect of various drug treatments on humoral alloimmunity. In order to satisfy this demand, we developed such a protocol in which de novo alloantibodies production is induced in one-way mixed lymphocyte reaction (MLR). The amount and capacity of the generated alloantibodies in the supernatant of each one-way MLR is assessed using an antibody-mediated cell dependent cytotoxicity (CDC) assay. The principle of the assay relies on the assessment of cellular survival of resting PBMCs isolated from the same donors, supplemented with the alloantibodies from the one-way MLR supernatant. The lesser is the cellular survival, the higher the production of alloantibodies in one-way MLR and consequently the more potent the humoral alloimmunity.
体液同种异体免疫仍然是一个重大且未解决的问题，它限制了同种异体移植物的存活。因此，需要开发一种简便、最好是非放射性且廉价的方案，用于评估各种药物治疗对体液同种异体免疫的影响。为了满足这一需求，我们开发了这样一种方案，即在单向混合淋巴细胞反应（MLR）中诱导产生新的同种异体抗体。使用抗体介导的细胞依赖性细胞毒性（CDC）测定法评估每个单向MLR上清液中产生的同种异体抗体的量和能力。该测定法的原理依赖于评估从相同供体分离的静息外周血单核细胞（PBMC）的细胞存活率，这些细胞补充了单向MLR上清液中的同种异体抗体。细胞存活率越低，单向MLR中同种异体抗体的产生就越高，因此体液同种异体免疫就越强。
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