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DISTINCT PATTERNS OF ENDOTHELIAL CELL ACTIVATION PRODUCED BY EXTRACELLULAR HISTONES AND BACTERIAL LIPOPOLYSACCHARIDE.

Sophia H Piffard1, Grant W Hennig, Adrian M Sackheim1

  • 1Departments of Emergency Medicine.

Shock (Augusta, Ga.)
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PubMed
Summary

Histones, not LPS, trigger rapid calcium responses in endothelial cells (ECs). Histones induce distinct calcium signaling patterns in arteries versus veins, highlighting differential EC activation mechanisms.

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Area of Science:

  • Cellular Biology
  • Vascular Biology
  • Immunology

Background:

  • Endothelial cells (ECs) respond to trauma factors like histone proteins and sepsis signals such as bacterial lipopolysaccharide (LPS) with calcium (Ca 2+ ) fluctuations.
  • The distinct patterns and kinetics of EC activation by histones versus LPS remain unclear.
  • Understanding these responses is crucial for differentiating cellular reactions to trauma and infection.

Purpose of the Study:

  • To investigate and compare the spatio-temporal patterns of Ca 2+ responses in ECs stimulated by histones and LPS.
  • To determine if histones elicit different Ca 2+ signaling patterns in arterial versus venous endothelial cells.
  • To explore the combined effects of LPS and histones on EC Ca 2+ activity and gene expression.

Main Methods:

  • High-speed live cell imaging of Ca 2+ events in cultured ECs (EA.hy926) and native ECs from murine arteries and veins.
  • Stimulation with histones or LPS, individually and in combination, followed by quantification of Ca 2+ prevalence.
  • RNA sequencing of ECs after 6-hour exposure to histones or LPS.

Main Results:

  • Histones rapidly increased Ca 2+ activity in ECs within seconds, while LPS had minimal effect.
  • Histones induced distinct, large aberrant Ca 2+ events in both arteries and veins, with unique spatio-temporal patterns ('rosettes' in arteries, individual cell responses in veins).
  • Pre-exposure to LPS potentiated histone-induced Ca 2+ activity; both stimuli altered gene expression, but with different induced mRNAs.

Conclusions:

  • Histones and LPS activate ECs via distinct, additive mechanisms, with histones uniquely causing significant aberrant Ca 2+ events.
  • Arterial and venous ECs exhibit differential spatio-temporal Ca 2+ responses to histone stimulation.
  • These findings reveal specific cellular responses to trauma and sepsis signals, impacting vascular health.