In vitro sepsis up-regulates Nociceptin/Orphanin FQ receptor expression and function on human T- but not B-cells.

Abstract

BACKGROUND AND PURPOSE:

In animal models of sepsis, increased activation of the Nociceptin/Orphanin FQ (N/OFQ) receptor NOP is associated with mortality and NOP antagonists improved survival. We have explored the role of the N/OFQ-NOP system in freshly isolated volunteer human B- and T-cells incubated with lipopolysaccharide (LPS) and peptidoglycan G (PepG) as a model of in vitro sepsis.

EXPERIMENTAL APPROACH:

B- and T-cell NOP expression was measured using the NOP fluorescent probe N/OFQATTO594 , N/OFQ content was measured using immunofluorescence, N/OFQ release was tracked using a CHOhNOPGαiq5 biosensor assay and NOP function was measured using transwell migration and cytokine/chemokine release using a 25-plex assay format. Cells were challenged with LPS/PepG.

KEY RESULTS:

CD19-positive B-cells bound N/OFQATTO594 ; they also contain N/OFQ. Stimulation with CXCL13/IL-4 increased N/OFQ release. N/OFQ trended to reduced migration to CXCL13/IL-4. Surface NOP expression was unaffected by LPS/PepG, but this treatment increased GM-CSF release in an N/OFQ sensitive manner. CD3-positive T-cells did not bind N/OFQATTO594 ; they did contain N/OFQ. Stimulation with CXCL12/IL-6 increased N/OFQ release. When incubated with LPS/PepG, NOP surface expression was induced leading to N/OFQATTO594 binding. In LPS/PepG-treated cells, N/OFQ reduced migration to CXCL12/IL-6. LPS/PepG increased GM-CSF release in an N/OFQ sensitive manner.

CONCLUSIONS AND IMPLICATIONS:

We suggest both a constitutive and sepsis-inducible N/OFQ-NOP receptor autocrine regulation of B- and T-cell function, respectively. These NOP receptors variably inhibit migration and reduce GM-CSF release. These data provide mechanistic insights to the detrimental role for increased N/OFQ signalling in sepsis and suggest a potential role for NOP antagonists as treatments.

• Authors: Mark F Bird, Christopher P Hebbes, Anushuya Tamang, Jonathon Mark Willets, Jonathan P Thompson, Remo Guerrini, Girolamo Calo, David G Lambert
• Journal: British journal of pharmacology (Br J Pharmacol) Vol. 180 Issue 17 Pg. 2298-2314 (09 2023) ISSN: 1476-5381 [Electronic] England
• PMID: 37021779 (Publication Type: Journal Article)
• Copyright: © 2023 The Authors. British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.

Chemical References

• nociceptin
• Receptors, Opioid
• Nociceptin Receptor
• Granulocyte-Macrophage Colony-Stimulating Factor
• Lipopolysaccharides
• Interleukin-4
• Interleukin-6
• Opioid Peptides

Topics

• Animals
• Humans
• Receptors, Opioid (metabolism)
• Nociceptin Receptor
• Granulocyte-Macrophage Colony-Stimulating Factor
• Lipopolysaccharides (pharmacology)
• Interleukin-4
• Interleukin-6
• Opioid Peptides (physiology)
• Sepsis (drug therapy)