Tissue-type plasminogen activator (tPA) is a major activator of fibrinolysis, which also attenuates the pro-inflammatory activity of lipopolysaccharide (LPS) in bone marrow-derived macrophages (BMDMs) and in vivo in mice. The activity of tPA as an LPS response modifier is independent of its proteinase activity and instead, dependent on the N-methyl-D-aspartate Receptor (NMDA-R), which is expressed by BMDMs. The major Toll-like receptor (TLR) for LPS is TLR4. Herein, we show that enzymatically-inactive (EI) tPA blocks the response of mouse BMDMs to selective TLR2 and TLR9 agonists, rapidly reversing IκBα phosphorylation and inhibiting expression of TNFα, CCL2, interleukin-1β, and interleukin-6. The activity of EI-tPA was replicated by activated α2-macroglobulin, which like EI-tPA, signals through an NMDA-R-dependent pathway. EI-tPA failed to inhibit cytokine expression by BMDMs in response to agonists that target the Pattern Recognition Receptors (PRRs), NOD1 and NOD2, providing evidence for specificity in the function of EI-tPA. Macrophages isolated from the peritoneal space (PMs), without adding eliciting agents, expressed decreased levels of cell-surface NMDA-R compared with BMDMs. These cells were unresponsive to EI-tPA in the presence of LPS. However, when PMs were treated with CSF-1, the abundance of cell-surface NMDA-R increased and the ability of EI-tPA to neutralize the response to LPS was established. We conclude that the anti-inflammatory activity of EI-tPA is selective for TLRs but not all PRRs. The ability of macrophages to respond to EI-tPA depends on the availability of cell surface NMDA-R, which may be macrophage differentiation-state dependent.

Tissue-type plasminogen activator selectively inhibits multiple toll-like receptors in CSF-1differentiated macrophages / Das, L.; Azmoon, P.; Banki, M. A.; Mantuano, E.; Gonias, S. L.. - In: PLOS ONE. - ISSN 1932-6203. - 14:11(2019), p. e0224738. [10.1371/journal.pone.0224738]

Tissue-type plasminogen activator selectively inhibits multiple toll-like receptors in CSF-1differentiated macrophages

Mantuano E.;
2019

Abstract

Tissue-type plasminogen activator (tPA) is a major activator of fibrinolysis, which also attenuates the pro-inflammatory activity of lipopolysaccharide (LPS) in bone marrow-derived macrophages (BMDMs) and in vivo in mice. The activity of tPA as an LPS response modifier is independent of its proteinase activity and instead, dependent on the N-methyl-D-aspartate Receptor (NMDA-R), which is expressed by BMDMs. The major Toll-like receptor (TLR) for LPS is TLR4. Herein, we show that enzymatically-inactive (EI) tPA blocks the response of mouse BMDMs to selective TLR2 and TLR9 agonists, rapidly reversing IκBα phosphorylation and inhibiting expression of TNFα, CCL2, interleukin-1β, and interleukin-6. The activity of EI-tPA was replicated by activated α2-macroglobulin, which like EI-tPA, signals through an NMDA-R-dependent pathway. EI-tPA failed to inhibit cytokine expression by BMDMs in response to agonists that target the Pattern Recognition Receptors (PRRs), NOD1 and NOD2, providing evidence for specificity in the function of EI-tPA. Macrophages isolated from the peritoneal space (PMs), without adding eliciting agents, expressed decreased levels of cell-surface NMDA-R compared with BMDMs. These cells were unresponsive to EI-tPA in the presence of LPS. However, when PMs were treated with CSF-1, the abundance of cell-surface NMDA-R increased and the ability of EI-tPA to neutralize the response to LPS was established. We conclude that the anti-inflammatory activity of EI-tPA is selective for TLRs but not all PRRs. The ability of macrophages to respond to EI-tPA depends on the availability of cell surface NMDA-R, which may be macrophage differentiation-state dependent.
2019
Animals; Cell Differentiation; Cytokines; Humans; Inflammation; Inflammation Mediators; Lipopolysaccharides; Macrophage Colony-Stimulating Factor; Macrophages; Mice, Inbred C57BL; Neutralization Tests; Nod1 Signaling Adaptor Protein; Nod2 Signaling Adaptor Protein; Receptors, N-Methyl-D-Aspartate; Tissue Plasminogen Activator; Toll-Like Receptors
01 Pubblicazione su rivista::01a Articolo in rivista
Tissue-type plasminogen activator selectively inhibits multiple toll-like receptors in CSF-1differentiated macrophages / Das, L.; Azmoon, P.; Banki, M. A.; Mantuano, E.; Gonias, S. L.. - In: PLOS ONE. - ISSN 1932-6203. - 14:11(2019), p. e0224738. [10.1371/journal.pone.0224738]
File allegati a questo prodotto
File Dimensione Formato  
Das_Tissue-type_2019.pdf

accesso aperto

Tipologia: Documento in Post-print (versione successiva alla peer review e accettata per la pubblicazione)
Licenza: Tutti i diritti riservati (All rights reserved)
Dimensione 1.79 MB
Formato Adobe PDF
1.79 MB Adobe PDF

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1403164
Citazioni
  • ???jsp.display-item.citation.pmc??? 9
  • Scopus 12
  • ???jsp.display-item.citation.isi??? 12
social impact