Lenart Marzena, Gruca Anna, Mueck Anna, Rutkowska-Zapała Magdalena, Surman Marta, Szaflarska Anna, Kobylarz Krzysztof, Baran Jarosław, Siedlar Maciej
Department of Clinical Immunology, Institute of Paediatrics, Faculty of Medicine, Jagiellonian University Medical College, Krakow, Poland.
Department of Clinical Immunology, University Children's Hospital, Krakow, Poland.
J Immunol Methods. 2017 Jul;446:1-6. doi: 10.1016/j.jim.2017.03.016. Epub 2017 Mar 30.
Invariant natural killer T (iNKT) cells are a small population of thymus-derived T cells that are restricted by non-classical MHC class I molecule CD1d and express an evolutionary conserved TCR with an invariant α-chain. The frequency of iNKT cells in peripheral blood is very low, thus, accurate methods to identify and enumerate iNKT cells are needed. The aim of the study was to compare 6B11 mAb or α-GalCer-loaded CD1d dextramers usage in iNKT cell detection. The frequency of CD3CD56 lymphocytes is much higher, with statistical significance (p<0,001), than real iNKT cells detected by 6B11 mAb or α-GalCer-loaded CD1d dextramers. The frequency of iNKT cells, recognized by 6B11 mAb or α-GalCer-loaded CD1d dextramers, was in a similar range. Nonetheless, when we compared whether 6B11 and α-GalCer-loaded CD1d dextramers are the same populations, it turned out that by this approach we were able to identify three distinct subsets of iNKT cells: i) 6B11/α-GalCer-loaded dextramer cells, ii) 6B11/α-GalCer-loaded dextramer cells, and iii) 6B11/α-GalCer-loaded dextramer. Thus, although 6B11 mAb and α-GalCer-loaded dextramers may identify not exactly the same cells, application of these methods seems to give similar results of iNKT cell frequency in peripheral blood. It seems that both approaches for iNKT detection can be used for precise identification of these cells. Moreover, our results indicate that CD3CD56 lymphocytes are a heterogeneous population of T cells, expressing activation markers of both NK and T lymphocytes, yet with not well characterized properties.
不变自然杀伤T（iNKT）细胞是一小部分源自胸腺的T细胞，受非经典MHC I类分子CD1d限制，并表达具有不变α链的进化保守TCR。外周血中iNKT细胞的频率非常低，因此，需要准确的方法来鉴定和计数iNKT细胞。本研究的目的是比较6B11单克隆抗体或负载α-半乳糖神经酰胺的CD1d二聚体在iNKT细胞检测中的应用。CD3CD56淋巴细胞的频率比用6B11单克隆抗体或负载α-半乳糖神经酰胺的CD1d二聚体检测到的真正iNKT细胞高得多，具有统计学意义（p<0.001）。用6B11单克隆抗体或负载α-半乳糖神经酰胺的CD1d二聚体识别的iNKT细胞频率在相似范围内。然而，当我们比较6B11和负载α-半乳糖神经酰胺的CD1d二聚体是否为同一群体时，结果表明通过这种方法我们能够鉴定出iNKT细胞的三个不同亚群：i）6B11/负载α-半乳糖神经酰胺的二聚体细胞，ii）6B11/负载α-半乳糖神经酰胺的二聚体细胞，iii）6B11/负载α-半乳糖神经酰胺的二聚体。因此，尽管6B11单克隆抗体和负载α-半乳糖神经酰胺的二聚体可能不能精确识别相同的细胞，但应用这些方法似乎在外周血中iNKT细胞频率方面给出相似的结果。似乎这两种iNKT检测方法都可用于这些细胞的精确鉴定。此外，我们的结果表明CD3CD56淋巴细胞是T细胞的异质群体，表达NK和T淋巴细胞的激活标志物，但其特性尚未得到很好的表征。
