Gupta-Wright Ankur, Tembo Dumizulu, Jambo Kondwani C, Chimbayo Elizabeth, Mvaya Leonard, Caldwell Shannon, Russell David G, Mwandumba Henry C
College of Medicine, Malawi-Liverpool-Wellcome Trust Clinical Research Programme, Blantyre, Malawi.
Clinical Research Department, London School of Hygiene and Tropical Medicine, London, United Kingdom.
Front Immunol. 2017 Sep 28;8:1222. doi: 10.3389/fimmu.2017.01222. eCollection 2017.
The accurate assessment of immune competence through analysis is paramount to our understanding of those immune mechanisms that lead to protection or susceptibility against a broad range of human pathogens. We have developed a flow cytometry-based, whole blood phagocyte functional assay that utilizes the inflammatory inducer zymosan, coupled to OxyBURST-SE, a fluorescent reporter of phagosomal oxidase activity. The assay measures both phagocytic uptake and the superoxide burst in the phagocyte populations in whole blood. We utilized this assay to demonstrate impaired superoxide burst activity in the phagocytes of hospitalized HIV-positive patients with laboratory-confirmed tuberculosis. These data validate the use of the assay to assess the immune competence of patients in a clinical setting. The method is highly reproducible with minimal intraindividual variation and opens opportunities for the rapid assessment of cellular immune competence in peripheral blood in a disease setting.
通过分析准确评估免疫能力对于我们理解导致对多种人类病原体产生保护或易感性的免疫机制至关重要。我们开发了一种基于流式细胞术的全血吞噬细胞功能测定法，该方法利用炎性诱导剂酵母聚糖，并与吞噬体氧化酶活性的荧光报告分子OxyBURST-SE偶联。该测定法可测量全血中吞噬细胞群体的吞噬摄取和超氧化物爆发。我们利用该测定法证明了实验室确诊为结核病的住院HIV阳性患者的吞噬细胞中超氧化物爆发活性受损。这些数据验证了该测定法在临床环境中用于评估患者免疫能力的用途。该方法具有高度可重复性，个体内变异极小，并为在疾病环境中快速评估外周血中的细胞免疫能力提供了机会。
