Shin Hye-Won, Umber Brandon J, Meinardi Simone, Leu Szu-Yun, Zaldivar Frank, Blake Donald R, Cooper Dan M
Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA.
J Transl Med. 2009 Apr 29;7:31. doi: 10.1186/1479-5876-7-31.
Noninvasive detection of innate immune function such as the accumulation of neutrophils remains a challenge in many areas of clinical medicine. We hypothesized that granulocytes could generate volatile organic compounds.
To begin to test this, we developed a bioreactor and analytical GC-MS system to accurately identify and quantify gases in trace concentrations (parts per billion) emitted solely from cell/media culture. A human promyelocytic leukemia cell line, HL60, frequently used to assess neutrophil function, was grown in serum-free medium.
HL60 cells released acetaldehyde and hexanaldehyde in a time-dependent manner. The mean +/- SD concentration of acetaldehyde in the headspace above the cultured cells following 4-, 24- and 48-h incubation was 157 +/- 13 ppbv, 490 +/- 99 ppbv, 698 +/- 87 ppbv. For hexanaldehyde these values were 1 +/- 0.3 ppbv, 8 +/- 2 ppbv, and 11 +/- 2 ppbv. In addition, our experimental system permitted us to identify confounding trace gas contaminants such as styrene.
This study demonstrates that human immune cells known to mimic the function of innate immune cells, like neutrophils, produce volatile gases that can be measured in vitro in trace amounts.
在临床医学的许多领域,对中性粒细胞聚集等先天免疫功能进行无创检测仍是一项挑战。我们推测粒细胞能够产生挥发性有机化合物。
为了开始验证这一点,我们开发了一种生物反应器和分析气相色谱 - 质谱系统,以准确识别和定量仅从细胞/培养基培养物中释放的痕量浓度(十亿分之一)的气体。一种常用于评估中性粒细胞功能的人早幼粒细胞白血病细胞系HL60,在无血清培养基中培养。
HL60细胞以时间依赖性方式释放乙醛和己醛。在培养4小时、24小时和48小时后,培养细胞上方顶空中乙醛的平均±标准差浓度分别为157±13 ppbv、490±99 ppbv、698±87 ppbv。对于己醛,这些值分别为1±0.3 ppbv、8±2 ppbv和11±2 ppbv。此外,我们的实验系统使我们能够识别苯乙烯等混杂的痕量气体污染物。
本研究表明,已知模拟先天免疫细胞(如中性粒细胞)功能的人类免疫细胞会产生挥发性气体,这些气体可在体外进行痕量测量。