细胞聚集是淋巴细胞对各种细胞外刺激的生理反应。目前,淋巴细胞聚集仅通过定性或半定量方法进行评估。在这项研究中,我们评估了流式细胞术定量测量淋巴细胞聚集的能力,准确,和可重复的方式,并检查了聚集反应在各种淋巴增生性疾病中的意义。
利用诸如抗CD19抗体或佛波醇酯的细胞外触发剂以浓度依赖性方式诱导淋巴细胞聚集。基于正向或侧向散射(SSC)通过流式细胞术定量聚集,或通过ImageStreamX测量的聚集体的暗场SSC。准确性,再现性,并对方法的局限性进行了评估。在各种类型的淋巴增生性疾病中测量了聚集反应,并与慢性淋巴细胞白血病的免疫表型和IGHV突变状态相关。
由细胞外刺激引起的淋巴样聚集体相对于每个事件中的细胞数量升高正向和SSC信号。聚集反应在不同类型的淋巴增生性疾病中有所不同。此外,CD19诱导的聚集水平升高与慢性淋巴细胞白血病的异常特征有关,但不是IGHV突变状态的疾病
我们已经证明,流式细胞术可以提供准确和可重复的测量初级以及T和B细胞系聚集响应细胞外刺激。使用活化驱动的或其他细胞聚集的定量评估可以提供分析工具来阐明与淋巴增殖性疾病相关的生化和分子机制。©2015国际临床细胞计数学会。
Cellular aggregation is a physiological response of lymphocytes to various extracellular stimuli. Currently, lymphocytes aggregation is only evaluated qualitatively or by semiquantitative methods. In this study, we assessed the capacity of flow cytometry to measure lymphocytes aggregation in a quantitative, accurate, and reproducible manner, and examined the significance of aggregation responses in various lymphoproliferative diseases.
Extracellular triggers such as anti-CD19 antibodies or phorbol ester were utilized to induce lymphoid cells aggregation in a concentration dependent manner. Aggregation was quantified by flow cytometry based on the forward or side scatter (SSC), or by dark-field SSC of aggregates measured by ImageStreamX. Accuracy, reproducibility, and limitations of the methodology were evaluated. Aggregation responses were measured in various types of lymphoproliferative diseases, and correlated with immunophenotyping and IGHV mutational status in chronic lymphocytic leukemia.
Lymphoid aggregates provoked by extracellular stimuli elevate the forward and SSC signals relatively to the number of cells in each event. Aggregation responses vary among different types of lymphoproliferative diseases. Moreover, elevated levels of CD19-induced aggregation are associated with aberrant chronic lymphocytic leukemia characteristics, but not with IGHV mutational status of the disease
We have demonstrated that flow cytometry can provide accurate and reproducible measurement of both primary as well as T and B cell lines aggregation in response to extracellular stimuli. The use of quantitative evaluation of activation driven or other cellular aggregation may provide an analytical tool to elucidate biochemical and molecular mechanisms associated with lymphoproliferative diseases. © 2015 International Clinical Cytometry Society.