Flow Cytometry Analysis of the Reactive Oxygen Species in Immature Granulocytes in Septic Patient

Sepsis Clinical Trial

— SEPSIROS  

Official title:

Flow Cytometry Analysis of the Reactive Oxygen Species in Immature Granulocytes in Septic Patient

Clinical Trial Details — Status: Terminated

Administrative data

• NCT number: NCT03846596
• Other study ID #: 87RI18-0031 (SEPSIROS)
• Status: Terminated
• Start date: March 26, 2019
• Est. completion date: March 10, 2020

Study information

• Verified date: April 2020
• Source: University Hospital, Limoges
• Contact: n/a
• Is FDA regulated: No
• Study type: Observational

Clinical Trial Summary

Sepsis is life-threatening organ dysfunction caused by a dysregulated host response to infection. Early during sepsis course, immature neutrophils could be found in the bloodstream and may be less efficient than mature neutrophils in reactive oxygen species (ROS) production. ROS induce an oxidative stress for bacteria which can protect through the SOS response. The main objective is to evaluate the level of ROS produced in the early steps of sepsis by the immature neutrophils.

Description:

Sepsis is life-threatening organ dysfunction caused by a dysregulated host response to infection. The immune system is activated by both pathogen-associated and host-derived molecular patterns. A strong response of neutrophils is engaged and both innate and adaptive immune system homeostasis are strongly affected. Neutrophils are able to produce high concentrations of inducible reactive oxygen species (ROS), leading to an oxidative stress. ROS can be released extracellularly at the site of infection or intracellularly in the phagolysosome. At early phase of sepsis, immature granulocytes are present in the bloodstream and could help to predict sepsis deterioration. However, it has also been shown that they are less efficient than mature granulocytes in ROS production and phagocytosis. ROS are potent stressors for bacteria and can directly or indirectly damage DNA. Bacteria can protect against DNA damage through the SOS response, which is a coordinated cellular response regulated by a repressor, LexA, and a sensor/activator, RecA. The bacterial SOS response is involved in acquisition of resistances to antibiotics through increasing frequencies of spontaneous mutations or increasing the expression of resistance and adaptation genes. The hypothesis that the low-level production of ROS by immature granulocytes in the early steps of sepsis could be beneficial for both the host, as a high level of ROS induce organ damage and dysfunction, and the pathogen, as low concentrations of ROS would be able to induce the SOS response allowing bacteria to enhance an adaptive response. The main objective it is to evaluate the level of ROS produced by the immature granulocytes in septic patient. Then, it will be assess if it could promote antibiotic resistance expression via SOS-induced integron gene cassette rearrangements.

Recruitment information / eligibility

• Status: Terminated
• Enrollment: 34
• Est. completion date: March 10, 2020
• Est. primary completion date: March 10, 2020
• Accepts healthy volunteers: No
• Gender: All
• Age group: 18 Years and older

Eligibility

Inclusion Criteria:

• Adult patients hospitalized in ICU or ED for acute sepsis

Exclusion Criteria:

• Immunosupressed patients

• Active cancer

• HIV

• Hematological or inflammatory diseases

Related Conditions & MeSH terms

• Sepsis

Intervention

Biological:
• Additional blood tube
Additional blood tube during care

Locations

Country	Name	City	State
France	Limoges university Hospital	Limoges

Sponsors (1)

Lead Sponsor	Collaborator
University Hospital, Limoges

Country where clinical trial is conducted

France, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	ROS level by Flow cytometry in immature granulocytes dosage sepsis	The biological variable obtained is an average of fluorescence intensity (MFI) corresponding to the amount of ROS contained in the immature granulocytes from the onset of sepsis to the acute phase	Day 1
Secondary	ROS level by Flow cytometry in mature granulocytes dosage sepsis	The biological variable obtained is an average of fluorescence intensity (MFI) corresponding to the amount of ROS contained in the mature granulocytes from the onset of sepsis to the acute phase	Day 1
Secondary	Bacterial phagocytosis capacity of granulocytes assess by flow cytometry	The biological variable obtained is an average of fluorescence intensity (MFI) corresponding to the number of ingested bacteria.	Day 1
Secondary	SOS response activation by bacteria determined by flow cytometry	The biological variable obtained is an average of fluorescence intensity (MFI) corresponding to the expression of the SOS gene by the bacteria.	Day 1
Secondary	Hospital mortality	number of death	Day 28
Secondary	ROS level by Flow cytometry in monocyte dosage sepsis	The biological variable obtained is an average of fluorescence intensity (MFI) corresponding to the amount of ROS contained in the monocyte from the onset of sepsis to the acute phase	Day 1
Secondary	ROS level by Flow cytometry in lymphocyte dosage sepsis	The biological variable obtained is an average of fluorescence intensity (MFI) corresponding to the amount of ROS contained in the lymphocyte from the onset of sepsis to the acute phase	Day 1