Characterization of Non-canonical Way in Inflammasome Monocytes of Patients With Severe Sepsis

Severe Sepsis Clinical Trial

— CASPASEs  

Official title:

Characterization of Non-canonical Way in Inflammasome Monocytes of Patients With Severe Sepsis

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT02539147
• Other study ID #: 2013-A01617-38
• Status: Completed
• Phase: N/A
• First received: August 21, 2015
• Last updated: February 20, 2017
• Start date: September 2014
• Est. completion date: December 2014

Study information

• Verified date: August 2015
• Source: Groupe Hospitalier Paris Saint Joseph
• Contact: n/a
• Is FDA regulated: No
• Study type: Observational

Clinical Trial Summary

Activation of caspase-4 and human caspase-5 (orthologs of caspase-11 in mice) in innate immune cells.

Description:

Lipopolysaccharide (LPS) of the wall of Gram-negative bacteria is one of pathogen associated molecular patterns (PAMPs), which are recognized by cells of the innate immune system via Toll-like receptor 4 (TLR4) . The LPS / TLR4 interaction induces the secretion of a variety of proinflammatory cytokines. Among them, interleukin-1β (IL-1β) is a major cytokine, and alterations to its secretion has been associated with various diseases, such as periodic syndrome associated cryopyrin (CAPS), gout, rheumatoid arthritis or multiple sclerosis.

The release of IL-1β is controlled by a molecular platform protein, known as the inflammasome name. The canonical protein (that is to say conserved in evolution) of the inflammasome NLRP3 are NLRC4 and that engages the ASC adapter protein to activate caspase-1, which promotes the cleavage of interleukin IL-1β. Gram-negative bacteria such as Escherichia coli cause non-canonical activation of caspase-1, caspase-11In involving more NLRP3 and AUC. In mouse, caspase-11 is essential to the immune response to gram-negative bacteria, but bacterial PAMPs that are responsible for triggering of the non-canonical inflammasome remain to be identified. It has recently been shown that caspase-11 is involved in the death of the mice subjected to septic shock. The mouse model requires caspase-11-dependent mechanism, caspase-independent -1 since the CASP1 - / - mouses are also susceptible to LPS as wild type mice. A key question to be answered is whether LPS triggers similar events in human cells. If this is the case, this knowledge may be useful in drug development for treatment of sepsis.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 6
• Est. completion date: December 2014
• Est. primary completion date: December 2014
• Accepts healthy volunteers: No
• Gender: All
• Age group: N/A and older

Eligibility

Inclusion Criteria:

• patients with sepsis severe (bacterial infection associated with deterioration of at least one vital function) defined according to the criteria consensus conference

• patients whose diagnosis is made within 24 hours of the opening hours of Pasteur Research Laboratory

• patients with arterial catheter (not to impose an additional puncture to the patient) will be recruited in the intensive care unit of the Hospital St. Joseph. Arterial blood (20 ml) will be charged within 24 hours of the onset of severe sepsis criteria, during working hours of the laboratory of the Pasteur Institute, using vacuum tubes under heparin (Vacutainer), sent by a courier within 2 hours following the removal (to ensure that the cells are sufficiently "fresh" during processing in the laboratory)

Exclusion Criteria:

Related Conditions & MeSH terms

• Sepsis
• Severe Sepsis
• Toxemia

Intervention

Biological:
• blood samples
Isolation and stimulation of monocytes, preparation of cell lysates, measurement of cytokine production, Western blot

Locations

Country	Name	City	State
France	Groupe Hospitalier Paris Saint Joseph	Paris	Ile de France

Sponsors (3)

Lead Sponsor	Collaborator
Groupe Hospitalier Paris Saint Joseph	Agency for Science, Technology and Research, Institut Pasteur

Country where clinical trial is conducted

France, 

References & Publications (5)

Bone RC, Balk RA, Cerra FB, Dellinger RP, Fein AM, Knaus WA, Schein RM, Sibbald WJ. Definitions for sepsis and organ failure and guidelines for the use of innovative therapies in sepsis. The ACCP/SCCM Consensus Conference Committee. American College of Chest Physicians/Society of Critical Care Medicine. Chest. 1992 Jun;101(6):1644-55. Review. — View Citation

Conforti-Andreoni C, Ricciardi-Castagnoli P, Mortellaro A. The inflammasomes in health and disease: from genetics to molecular mechanisms of autoinflammation and beyond. Cell Mol Immunol. 2011 Mar;8(2):135-45. doi: 10.1038/cmi.2010.81. Review. — View Citation

Kayagaki N, Warming S, Lamkanfi M, Vande Walle L, Louie S, Dong J, Newton K, Qu Y, Liu J, Heldens S, Zhang J, Lee WP, Roose-Girma M, Dixit VM. Non-canonical inflammasome activation targets caspase-11. Nature. 2011 Oct 16;479(7371):117-21. doi: 10.1038/nature10558. — View Citation

Viganò E, Mortellaro A. Caspase-11: the driving factor for noncanonical inflammasomes. Eur J Immunol. 2013 Sep;43(9):2240-5. doi: 10.1002/eji.201343800. Review. — View Citation

Zambetti LP, Laudisi F, Licandro G, Ricciardi-Castagnoli P, Mortellaro A. The rhapsody of NLRPs: master players of inflammation...and a lot more. Immunol Res. 2012 Sep;53(1-3):78-90. doi: 10.1007/s12026-012-8272-z. Review. — View Citation

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Characterize the expression of caspase-4 and -5 in by measuring the production of cytokines: IL-1ß concentrations and IL-1a will determined by ELISA in the cell supernatant will be collected from monocytes stimulated or not		Two hours after blood collection : isolation and stimulation of monocytes than preparation of cell lysates
Secondary	Correlate the activation of caspases-4 and -5 with IL-1ß production with (canonical inflammasome) and IL-1a (Non-canonical inflammasome) measured by Western blot		Two hours after blood collection : isolation and stimulation of monocytes than preparation of cell lysates