Smith Tracey J, Wilson Marques A, Young Andrew J, Montain Scott J
Military Nutrition Division, United States Army Research Institute of Environmental Medicine, Natick, MA, United States.
J Immunol Methods. 2015 Feb;417:124-130. doi: 10.1016/j.jim.2015.01.002. Epub 2015 Jan 10.
Skin wound healing models can be used to detect changes in immune function in response to interventions. This study used a test-retest format to assess the reliability of a skin suction blister procedure for quantitatively evaluating human immune function in repeated measures type studies. Up to eight suction blisters (~30 mm(2)) were induced via suction on each participant's left and right forearm (randomized order; blister session 1 and 2), separated by approximately one week. Fluid was sampled from each blister, and the top layer of each blister was removed to reveal up to eight skin wounds. Fluid from each wound was collected 4, 7 and 24h after blisters were induced, and proinflammatory cytokines were measured. Transepidermal water loss (TEWL), to assess skin barrier recovery, was measured daily at each wound site until values were within 90% of baseline values (i.e., unbroken skin). Sleep, stress and inflammation (i.e., factors that affect wound healing and immune function), preceding the blister induction, were assessed via activity monitors (Actical, Philips Respironics, Murrysville, Pennsylvania), the Perceived Stress Scale (PSS) and C-reactive protein (CRP), respectively. Area-under-the-curve and TEWL, between blister session 1 and 2, were compared using Pearson correlations and partial correlations (controlling for average nightly sleep, PSS scores and CRP). The suction blister method was considered reliable for assessing immune response and skin barrier recovery if correlation coefficients reached 0.7. Volunteers (n=16; 12 M; 4F) were 23 ± 5 years [mean ± SD]. Time to skin barrier restoration was 4.9 ± 0.8 and 4.8 ± 0.9 days for sessions 1 and 2, respectively. Correlation coefficients for skin barrier restoration, IL-6, IL-8 and MIP-1α were 0.9 (P<0.0001), 0.7 (P=0.008) and 0.9 (P<0.0001), respectively. When average nightly sleep, PSS scores and CRP (i.e., percent difference between sessions 1 and 2) were taken into consideration, correlations in immune response between sessions 1 and 2 were improved for IL-8 (0.8, P=0.002) and TNF-α (0.7, P=0.02). The skin suction blister method is sufficiently reliable for assessing skin barrier restoration and immune responsiveness. This data can be used to determine sample sizes for cross-sectional or repeated-measures types of studies testing the impact of various stressors on immune response, and/or the efficacy of interventions to mitigate decrements in immune response to stress.
皮肤伤口愈合模型可用于检测免疫功能在干预后的变化。本研究采用重复测量的方式，评估皮肤抽吸水疱法在重复测量类型研究中定量评估人体免疫功能的可靠性。在每位参与者的左右前臂上通过抽吸诱导产生多达8个抽吸水疱（约30平方毫米）（随机顺序；水疱实验1和2），两次实验间隔约一周。从每个水疱中采集液体，并去除每个水疱的顶层以暴露出多达8个皮肤伤口。在诱导水疱后4小时、7小时和24小时收集每个伤口的液体，并测量促炎细胞因子。每天在每个伤口部位测量经表皮水分流失（TEWL）以评估皮肤屏障恢复情况，直至数值达到基线值（即完整皮肤）的90%以内。在水疱诱导前，分别通过活动监测仪（Actical，飞利浦伟康公司，宾夕法尼亚州默里斯维尔）、感知压力量表（PSS）和C反应蛋白（CRP）评估睡眠、压力和炎症（即影响伤口愈合和免疫功能的因素）。使用Pearson相关性和偏相关性（控制平均夜间睡眠、PSS评分和CRP）比较水疱实验1和2之间的曲线下面积和TEWL。如果相关系数达到0.7，则认为皮肤抽吸水疱法在评估免疫反应和皮肤屏障恢复方面是可靠的。志愿者（n = 16；12名男性；4名女性）年龄为23±5岁[平均值±标准差]。实验1和2中皮肤屏障恢复的时间分别为4.9±0.8天和4.8±0.9天。皮肤屏障恢复、IL - 6、IL - 8和MIP - 1α的相关系数分别为0.9（P < 0.0001）、0.7（P = 0.008）和0.9（P < 0.0001）。当考虑平均夜间睡眠、PSS评分和CRP（即实验1和2之间的百分比差异）时，实验1和2之间IL - 8（0.8，P = 0.002）和TNF - α（0.7，P = 0.02）的免疫反应相关性得到改善。皮肤抽吸水疱法在评估皮肤屏障恢复和免疫反应性方面具有足够的可靠性。这些数据可用于确定横断面或重复测量类型研究的样本量，这些研究旨在测试各种应激源对免疫反应的影响，和/或减轻应激导致的免疫反应下降的干预措施的疗效。
