Nie Shuai, Zhang Huaibin, Mayer Kathryn M, Oppenheim Frank G, Little Frédéric F, Greenberg Jonathan, Uluer Ahmet Z, Walt David R
Department of Chemistry, Tufts University, Medford, Massachusetts, United States of America.
Goldman School of Dental Medicine, Boston University, Boston, Massachusetts, United States of America.
PLoS One. 2015 Aug 10;10(8):e0135237. doi: 10.1371/journal.pone.0135237. eCollection 2015.
Monitoring clinical disease status in cystic fibrosis frequently requires invasive collection of clinical samples. Due to its noninvasive collection process and direct anatomic relationship with the lower airway, saliva shows great potential as a biological fluid for cystic fibrosis monitoring.
To measure the levels of multiple protein markers in human saliva supernatants and investigate the possibility of utilizing them to provide a more quantitative measure of disease state for use in research and monitoring of patients with cystic fibrosis clinically.
Whole saliva samples were collected and processed from cystic fibrosis patients at two distinct time points (2010 and 2013) and measured by two separate platforms. In this cross sectional study, a convenience sample of 71 participants were recruited with samples measured by multiplexed fluorescence microarray (fiber microarray) and another 117 participant samples were measured by an automated, point-of-care, analyzer (SDReader) using a microsphere-based array via fluorescence sandwich immunoassay. For comparison, saliva from 56 and 50 healthy subjects were collected, respectively. The levels of six target proteins were quantified. Various demographic and clinical data, including spirometry, medical history, and clinicians' assessments were also collected from patients with cystic fibrosis on the day of saliva collection.
Similar trends were observed with both platforms and compared with healthy subjects, cystic fibrosis patients had significantly elevated levels of VEGF, IP-10, IL-8, and EGF as well as lower levels of MMP-9 (P ≤ 0.005) using fiber microarray and significantly elevated levels of IP-10, IL-8 with lower levels of MMP-9 and IL-1β (P ≤ 0.02) using the SDReader. The levels of the six proteins correlated with each other significantly, and in some cases, biomarker levels could be used to differentiate between subgroups of patients with different clinical presentations. For example, IP-10 levels significantly correlated with FEV1 and disease severity (as evaluated by clinicians) with both platforms (P < 0.05).
Significant variations of the levels of six proteins in saliva supernatants, and the correlations of these levels with clinical assessments, demonstrated the potential of saliva for cystic fibrosis research and monitoring.
监测囊性纤维化的临床疾病状态通常需要通过侵入性手段采集临床样本。由于唾液的采集过程是非侵入性的，且与下呼吸道存在直接的解剖学关系，因此唾液作为一种生物流体在囊性纤维化监测方面具有巨大潜力。
测量人唾液上清液中多种蛋白质标志物的水平，并研究利用这些标志物更定量地评估疾病状态的可能性，以便用于囊性纤维化患者的临床研究和监测。
在两个不同时间点（2010年和2013年）收集囊性纤维化患者的全唾液样本并进行处理，通过两个不同平台进行测量。在这项横断面研究中，招募了71名参与者的便利样本，其样本通过多重荧光微阵列（纤维微阵列）进行测量，另外117名参与者的样本通过基于微球阵列的荧光夹心免疫测定法，使用自动化即时检测分析仪（SDReader）进行测量。为作比较，分别收集了56名和50名健康受试者的唾液。对六种目标蛋白的水平进行了定量分析。在采集唾液当天，还从囊性纤维化患者处收集了各种人口统计学和临床数据，包括肺功能测定、病史和临床医生的评估。
两个平台均观察到相似趋势，与健康受试者相比，使用纤维微阵列时，囊性纤维化患者的血管内皮生长因子（VEGF）、γ干扰素诱导蛋白10（IP - 10）、白细胞介素 - 8（IL - 8）和表皮生长因子（EGF）水平显著升高，基质金属蛋白酶 - 9（MMP - 9）水平较低（P≤0.005）；使用SDReader时，IP - 10、IL - 8水平显著升高，MMP - 9和白细胞介素 - 1β（IL - 1β）水平较低（P≤0.02）。六种蛋白质的水平之间存在显著相关性，在某些情况下，生物标志物水平可用于区分具有不同临床表现的患者亚组。例如，两个平台的IP - 10水平均与第一秒用力呼气容积（FEV1）和疾病严重程度（由临床医生评估）显著相关（P < 0.05）。
唾液上清液中六种蛋白质水平存在显著差异，且这些水平与临床评估相关，这表明唾液在囊性纤维化研究和监测方面具有潜力。
