Journal of Molecular Biomarkers & Diagnosis

Gas Signatures from Cultured Neutrophils and Peripheral Blood Mononuclear Cells Obtained from Healthy Humans

Abstract

Hye-Won Shin, Brandon J. Umber, Simone Meinardi, Szu-Yun Leu, Frank Zaldivar, Donald R. Blake and Dan M. Cooper

We hypothesized that leucocytes may be a significant source of physiologically active gases. Furthermore, it is hoped that alterations by disease of these volatile organic compound (VOC) profiles can ultimately be used as noninvasive biomarkers. We recently demonstrated that transformed promyelocitic cells produce distinct gases. (J Transl Med. 2009 7:31). The current study extends this work to neutrophils and peripheral blood mononuclear cells (PBMCs). Cells were isolated from the peripheral blood of healthy donors (n=10, 18-65 yrs old), resuspended in RPMI, and incubated in bioreactors for 24 hrs. The headspace was analyzed using gas chromatography. Acetaldehyde was elevated from neutrophils(median (min, max); 197 (34, 577) ppbv) compared with media (88 (40,116) ppbv, p=0.014). In the presence of alanine, neutrophils emitted more acetaldehyde (>1.5 fold compared to basal level). In contrast, acetaldehyde from PBMCs was 26 (8, 89) ppbv, significantly below media (p=0.004). Adding alanine did not affect acetaldehyde emissions from PBMCs. Also, hexanaldehyde appeared metabolized by neutrophils and PBMCs. This study demonstrates 1) that human primary immune cells produce measurable VOCs in vitro, and 2) the ability to detect basal levels of acetaldehyde from unperturbed cultured neutrophils. Moreover, the data suggest that different leukocyte subtypes have different VOC profiles.