1. Infrared spectroscopic characterization of monocytic microvesicles (microparticles) released upon lipopolysaccharide stimulation
- Author
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Lee, J, Wen, B, Carter, EA, Combes, V, Grau, GER, and Lay, PA
- Subjects
Lipopolysaccharides ,Biochemistry & Molecular Biology ,Cell-Derived Microparticles ,Spectroscopy, Fourier Transform Infrared ,Humans ,Flow Cytometry ,Monocytes ,Cell Line - Abstract
© FASEB. Microvesicles (MVs) are involved in cell-cell interactions, including disease pathogenesis. Nondestructive Fourier-transform infrared (FTIR) spectra from MVs were assessed as a technique to provide new biochemical insights into a LPS-induced monocyte model of septic shock. FTIR spectroscopy provided a quick method to investigate relative differences in biomolecular content of different MV populations that was complementary to traditional semiquantitative omics approaches, with which it is difficult to provide information on relative changes between classes (proteins, lipids, nucleic acids, carbohydrates) or protein conformations. Timedependent changes were detected in biomolecular contents of MVs and in the monocytes from which they were released. Differences in phosphatidylcholine and phosphatidylserine contents were observed in MVs released under stimulation, and higher relative concentrations of RNA and a-helical structured proteins were present in stimulatedMVs compared withMVs fromresting cells. FTIR spectra of stimulated monocytes displayed changes that were consistent with those observed in the correspondingMVs they released. LPS-stimulated monocytes had reduced concentrations of nucleic acids, a-helical structured proteins, and phosphatidylcholine compared with restingmonocytes but had an increase in total lipids. FTIRspectra ofMVbiomolecular content will be important in shedding new light on the mechanisms of MVs and the different roles they play in physiology and disease pathogenesis.
- Published
- 2017