Clinical trial NCT03846596: Flow Cytometry Analysis of the Reactive Oxygen Species in Immature Granulocytes in Septic Patient
Official title: Flow Cytometry Analysis of the Reactive Oxygen Species in Immature Granulocytes in Septic Patient
Study type: OBSERVATIONAL
Overall status: TERMINATED
Conditions: Sepsis
Keywords: Sepsis
reactive oxygen species
immature neutrophils
Lead sponsor: University Hospital, Limoges
Sponsor class: OTHER
First posted: 2019-02-19
Enrollment: 34
Sex: ALL
Minimum age (years): 18
Standard ages: ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Observational model: COHORT
Time perspective: PROSPECTIVE
Sampling method: PROBABILITY_SAMPLE
Study population: Patient hospitalized in intensive car or emergency department for acute sepsis

Brief 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.

Detailed 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.

Eligibility criteria: Inclusion Criteria:

• Adult patients hospitalized in ICU or ED for acute sepsis

Exclusion Criteria:

* Immunosupressed patients
* Active cancer
* HIV
* Hematological or inflammatory diseases

Primary outcomes:
measure: ROS level by Flow cytometry in immature granulocytes dosage sepsis
timeFrame: Day 1
Secondary outcomes:
measure: ROS level by Flow cytometry in mature granulocytes dosage sepsis
timeFrame: Day 1
measure: Bacterial phagocytosis capacity of granulocytes assess by flow cytometry
timeFrame: Day 1
measure: SOS response activation by bacteria determined by flow cytometry
timeFrame: Day 1
measure: Hospital mortality
timeFrame: Day 28
measure: ROS level by Flow cytometry in monocyte dosage sepsis
timeFrame: Day 1
measure: ROS level by Flow cytometry in lymphocyte dosage sepsis
timeFrame: Day 1
Other outcomes:
Locations:
city: Limoges
country: France
facility: Limoges university Hospital
Study officials:
affiliation: University Hospital, Limoges
name: Thomas DAIX, MD
role: PRINCIPAL_INVESTIGATOR
References: