质谱流式技术实现在单细胞水平检测几十个通道的同时检测,可以系统全面的分析病理状态下各类免疫细胞在表型和功能上的变化,从中发现一些新的免疫细胞亚群、揭示新的免疫机制。近年来,越来越多的实验室将质谱流式用于HIV的相关的研究,至2020年年初,已经有二十余篇文章发表,主要涉及HIV病毒感染不同阶段免疫系统的变化以及ART治疗相关免疫机制等方面。
01
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HIV病毒感染过程中对于不同免疫亚群的影响
1)CD4+ T
图1、HIV感染会造成CD4+T表型的显著改变
*注意B图中,由于几个主要marker的表达发生较大改变,在tsne图中,相当一部分被感染细胞(蓝色)已经无法和未感染(红色)细胞重合。图片来源:[4]
图2、tSNE显示HIV患者和健康对照体内TFH亚群的表型存在很大差异
此外,质谱流式还可以帮助研究者发现一些疾病相关的新亚群,揭示发病过程一些新的免疫机制。2019年Del Alcalza D等人利用质谱流式检测了HIV感染的淋巴结T细胞上36个marker的表达,系统分析了CD4+T细胞的亚群组成。研究发现,患者淋巴结内存在一群可以分泌IL-21的CXCR5-细胞。后续的细胞功能试验和TCR受体测序结果说明其具有和TFH类似的功能,并具有部分重合的TCR克隆谱系,同时该群细胞对外周血CXCR5-CD4+T cells也有贡献。这些信息串在一起,揭示了一个在HIV慢性感染过程中连接淋巴结病理学和循环T细胞的新机制,拓展了我们对于T调节B细胞功能多样性的认识。[5]。
2)CD8+ T
耗竭型CD8+T细胞是HIV进入慢性感染的标志。2018年Bengsch等人利用质谱流式对患者耗竭型CD8+T(Tex)的表型进行了深入研究。基于前期表观遗传的数据,作者筛选了16多个与功能耗竭相关marker加入质谱流式Panel进行分析,从单细胞的水平分析Tex的耗竭表型。经过PhenoGraph聚类,他们得到了在9个表型不同的Tex细胞亚群,定义了不同抑制性受体在各个Tex亚群上的表达组合模式。然后,根据他们的比例信息,制作了表征疾病状态的指标。这对于研究慢性感染,自身免疫和癌症中的免疫监测和免疫调节机制都具有重要意义(如图3)。[1]
图3 :质谱流式可以帮助表征疾病状态的指标或者生物标志物
(图片来源文献[1])
3)其他免疫细胞
02
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揭示在ART治疗过程中免疫细胞机制研究
小结:
综上,通过单细胞水平的多参数检测,质谱流式可以全面系统的分析分析HIV病毒感染不同阶段以及治疗过程中免疫系统的表型和功能改变,为人类进一步了解和控制该疾病提供了有益的探索。在肿瘤等其他疾病的研究中,科研人员已经通过质谱流式找到了一些用于临床诊断和预后的生物标志物[22,23],相信未来在HIV的研究领域,还将有更多的更加贴近临床的研究和发现。
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