Infrared spectroscopic characterization of monocytic microvesicles (microparticles) released upon lipopolysaccharide stimulation.
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. Time-dependent 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 α-helical structured proteins were present in stimulated MVs compared with MVs from resting cells. FTIR spectra of stimulated monocytes displayed changes that were consistent with those observed in the corresponding MVs they released. LPS-stimulated monocytes had reduced concentrations of nucleic acids, α-helical structured proteins, and phosphatidylcholine compared with resting monocytes but had an increase in total lipids. FTIR spectra of MV biomolecular content will be important in shedding new light on the mechanisms of MVs and the different roles they play in physiology and disease pathogenesis.-Lee, J., Wen, B., Carter, E. A., Combes, V., Grau, G. E. R., Lay, P. A. Infrared spectroscopic characterization of monocytic microvesicles (microparticles) released upon lipopolysaccharide stimulation.
School of Chemistry and Vibrational Spectroscopy Core Facility, The University of Sydney, Sydney, New South Wales, Australia.
Joonsup
J
Lee
Vascular Immunopathology Unit, Bosch Institute-School of Medical Sciences, and.
Beryl
B
Wen
School of Chemistry and Vibrational Spectroscopy Core Facility, The University of Sydney, Sydney, New South Wales, Australia.
Elizabeth A
EA
Carter
Vascular Immunopathology Unit, Bosch Institute-School of Medical Sciences, and.|Faculty of Science, School of Life Sciences, University of Technology Sydney, Sydney, New South Wales, Australia.
Valery
V
Combes
Vascular Immunopathology Unit, Bosch Institute-School of Medical Sciences, and.|Australian Institute of Nanoscale Science and Technology (AINST), The University of Sydney, Sydney, New South Wales, Australia.
Georges E R
GER
Grau
School of Chemistry and Vibrational Spectroscopy Core Facility, The University of Sydney, Sydney, New South Wales, Australia; peter.lay@sydney.edu.au.|Australian Institute of Nanoscale Science and Technology (AINST), The University of Sydney, Sydney, New South Wales, Australia.
Peter A
PA
Lay
D002460:Cell Line; D055252:Cell-Derived Microparticles / Q000502:physiology*; D005434:Flow Cytometry; D006801:Humans; D008070:Lipopolysaccharides / Q000633:toxicity*; D009000:Monocytes / Q000187:drug effects* / Q000502:physiology*; D017550:Spectroscopy, Fourier Transform Infrared
D008070:Lipopolysaccharides
FTIR spectroscopy; cell signaling; endotoxin; extracellular vesicles; immunology