Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109; and.
Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington School of Medicine, Seattle, WA 98195.
J Immunol. 2020 Oct 15;205(8):2312-2320. doi: 10.4049/jimmunol.2000320. Epub 2020 Sep 14.
Measurements of IgG and IgA in human rectal secretions are used to evaluate the Abs elicited by HIV vaccines or the bioaccumulation following immunoprophylaxis at the sites of HIV exposure. To improve sampling methods and tolerability of the procedure, we optimized a balloon device (OriCol) for rectal microbiome sampling requiring 10 second inflation and compared this method to a 5 minute collection using sponges. Lubrication of the device did not interfere with IgG, IgA, or hemoglobin ELISA. Lubricated OriCols inflated to 30 cc minimized hemoglobin contamination (<4.68 ng/ml) compared with collections with two sponge types (Weck-Cel: 267.2 ng/ml, < 0.0001; and Merocel: 59.38 ng/ml, = 0.003). Median human serum albumin for OriCols was 14.9 μg/ml, whereas Merocels and Weck-Cels were 28.57 μg/ml ( = 0.0005) and 106.2 μg/ml ( = 0.0002), respectively. Consistent with reduced systemic contamination, the median IgG measured in OriCol-collected rectal secretions (986 ng) was lower than secretions from sponges (Weck-Cel: 8588 ng, < 0.0001; Merocel: 2509 ng, = 0.0389). The median IgA yield of samples using the OriCol method (75,253 ng) was comparable to that using Merocel (71,672 ng; = 0.6942) but significantly higher than Weck-Cel sponges (16,173 ng, = 0.0336). Median recovery volumes for OriCols were 800 μl, whereas Merocels and Weck-Cels were 615 μl ( = 0.0010) and 655 μl ( = 0.0113), respectively. The balloon device was acceptable among 23 participants, as 85.1% experiencing their first collection ranked it as "seven: acceptable - a lot" or "six: acceptable - somewhat" in a seven-point Likert scale. Therefore, lubricated OriCols inflated to 30 cc allowed for a rapid, well-tolerated, blood-free collection of human rectal secretions.
测量人类直肠分泌物中的 IgG 和 IgA 可用于评估 HIV 疫苗引起的抗体,或在 HIV 暴露部位进行免疫预防后的生物蓄积。为了改进采样方法并提高该程序的耐受性,我们优化了一种用于直肠微生物组采样的气球装置(OriCol),该装置需要充气 10 秒,并将该方法与使用海绵进行 5 分钟采集进行了比较。该装置的润滑不会干扰 IgG、IgA 或血红蛋白 ELISA。与两种海绵类型(Weck-Cel:267.2ng/ml,<0.0001;和 Merocel:59.38ng/ml,=0.003)相比,用 30cc 充气的润滑 OriCol 最小化了血红蛋白污染(<4.68ng/ml)。OriCol 的人血清白蛋白中位数为 14.9μg/ml,而 Merocel 和 Weck-Cel 分别为 28.57μg/ml(=0.0005)和 106.2μg/ml(=0.0002)。与系统性污染减少一致,OriCol 收集的直肠分泌物中测量的 IgG 中位数(986ng)低于海绵收集的分泌物(Weck-Cel:8588ng,<0.0001;Merocel:2509ng,=0.0389)。使用 OriCol 方法获得的 IgA 中位数产量(75253ng)与使用 Merocel(71672ng;=0.6942)相当,但明显高于 Weck-Cel 海绵(16173ng,=0.0336)。OriCol 的中位回收体积为 800μl,而 Merocel 和 Weck-Cel 分别为 615μl(=0.0010)和 655μl(=0.0113)。在 23 名参与者中,该气球装置是可以接受的,85.1%的首次使用者将其评为七点李克特量表中的“七:可接受-非常”或“六:可接受-有些”。因此,润滑后的 OriCol 充气至 30cc 可快速、耐受良好、无血采集人类直肠分泌物。