MedPath

Study of the Sex Differences in Inflammatory Diseases in Children

Not Applicable
Conditions
Acute Inflammatory Diseases in Children
Sex Differences in Immune Response
Interventions
Other: Blood collection
Other: Stool collection
Registration Number
NCT04815811
Lead Sponsor
Queen Fabiola Children's University Hospital
Brief Summary

Sexual differences in innate immune response have been demonstrated and were mainly attributed to the influence of the sex steroids (1-18). However, recent clinical data revealed significant differences in inflammatory markers between boys and girls suffering from acute and chronic inflammatory diseases (19-23). Sex hormone levels in prepubertal children are particularly low and insufficient to explain the gender differences observed in inflammatory conditions from neonates to the elderly, suggesting the contribution of another mechanism, such as the influence of genes situated on the sex chromosomes and involved in the inflammatory response.

The aim of this work is to evaluate the role of the X chromosome in the sex differences in inflammatory diseases in children. In order to discriminate more precisely the role of the X chromosome relatively to the sex steroids in the sex-specific inflammatory response, some innate immune functions related to X-linked genes will be evaluated in whole blood from prepubertal children of both sexes, suffering from acute inflammatory processes such as pyelonephritis caused by Escherichia coli, pneumonia with pleural effusion caused by Streptococcus pneumoniae or sepsis

Detailed Description

Many studies demonstrated immune differences between men and women suffering from acute and chronic inflammatory processes. In cases of acute inflammatory diseases, such as sepsis, females have better prognosis comparing to males (1,24-28).

On the contrary, worse prognosis for women is observed in chronic inflammatory diseases such as asthma or cystic fibrosis (8-10,12,13,29).

Sex-depended inflammatory response was attributed to the influence of sex hormones on the immune system. (2,15-18). However recent studies revealed differences in the clinical outcome but also in inflammatory markers between boys and girls suffering from acute and chronic inflammatory diseases (19-23). Sex hormone levels in prepubertal children are particularly low and insufficient to explain the gender differences observed in inflammatory conditions from neonates to the elderly, suggesting the contribution of another mechanism, such as the influence of genes situated on the sex chromosomes and involved in the inflammatory response.

The aim of this work is to identify the potential X-linked mechanisms responsible for some of the differences between boys and girls in the inflammatory response, making the girls more at risk of developing complications in chronic inflammatory diseases and the boys more at risk of lethal complications in severe acute inflammatory diseases like sepsis. Several genes coding for innate immunity components are linked to the X chromosome such as diapedesis molecule CD99 or TLR pathway proteins genes. (30-33). X chromosome is also highly enriched in genes encoding micro RNAs (miRNAs) involved in the post-transcriptional regulation of gene expression which play a critical role in immune inflammatory response (34-36).

Thus, in order to discriminate more precisely the role of the X chromosome relatively to the sex steroids in the sex-specific inflammatory response, some innate immune functions related to X-linked genes will be evaluated in whole blood from prepubertal children of both sexes, suffering from acute inflammatory processes such as pyelonephritis caused by Escherichia coli, pneumonia with pleural effusion caused by Streptococcus pneumoniae or sepsis. We will also study the correlations between inflammatory and clinical markers of the disease activity to identify prognosis indicators depending on the sex. Additionally, to delineate microbiome contribution, we will study the gut microbiota in stool samples obtained from the recruited patients.

Recruitment & Eligibility

Status
UNKNOWN
Sex
All
Target Recruitment
160
Inclusion Criteria

Not provided

Exclusion Criteria
  • Use of antithrombotic drugs (acetylsalicylic acid, thienopyridines, dipyridamol, glycoprotein IIb / IIIa antagonists, vitamin K antagonists, heparins).
  • Congenital or acquired immunodeficiency: immunosuppressive drugs, hematopoietic stem cells transplantation, immunoglobulin therapy, extracorporeal membrane oxygenation (ECMO).
  • Hemodialysis.
  • 48h following cardiac operation of any type.
  • Malignant cancer.
  • HIV.

Study & Design

Study Type
INTERVENTIONAL
Study Design
PARALLEL
Arm && Interventions
GroupInterventionDescription
Children suffering from acute inflammatory processes.Blood collectionThe study population will consist of male and female children, aged from 6 months to 7 years old, admitted to the hospital for one of the three following types of acute inflammatory processes: * Urinary tract infection caused by Escherichia coli * Pneumonia with pleural effusion caused by Streptococcus pneumoniae * Sepsis
Children suffering from acute inflammatory processes.Stool collectionThe study population will consist of male and female children, aged from 6 months to 7 years old, admitted to the hospital for one of the three following types of acute inflammatory processes: * Urinary tract infection caused by Escherichia coli * Pneumonia with pleural effusion caused by Streptococcus pneumoniae * Sepsis
Control groupBlood collectionMale and female children, aged from 6 months to 7 years old, admitted to the hospital for a scheduled operation for a non-inflammatory pathology.
Primary Outcome Measures
NameTimeMethod
Whole blood production of cytokine IL-6within 24 hours of hospital admission (Day 0)

The production of IL6 is measured by multiplex techniques.

Secondary Outcome Measures
NameTimeMethod
Intracellular quantity of the phosphorylated forms of NF-κB p65 in leukocyte population.within 24 hours of hospital admission (Day 0)
Intracellular quantity of the phosphorylated forms of ERK1/2 in leukocyte population.within 24 hours of hospital admission (Day 0)
Whole blood production of cytokine TNF-αwithin 24 hours of hospital admission (Day 0)
Whole blood production of cytokine interferon-αwithin 24 hours of hospital admission (Day 0)
Whole blood production of cytokine IL-1βwithin 24 hours of hospital admission (Day 0)
Whole blood production of cytokine IL-8within 24 hours of hospital admission (Day 0)
Whole blood production of cytokine IL-10within 24 hours of hospital admission (Day 0)
Intracellular quantity of the phosphorylated forms of p38 MAPK in leukocyte population.within 24 hours of hospital admission (Day 0)
Expression of the cell diapedesis receptor CD99 on monocyteswithin 24 hours of hospital admission (Day 0)

Measurements of cell diapedesis receptor CD99 on leukocytes will be performed by flow cytometry

IGF1within 24 hours of hospital admission (Day 0)
Expression of the cell diapedesis receptor CD99 on PMNswithin 24 hours of hospital admission (Day 0)

Measurements of cell diapedesis receptor CD99 on leukocytes will be performed by flow cytometry

Expression of TLR2 on monocyteswithin 24 hours of hospital admission (Day 0)

Measurements of intracellular phosphorylated forms of TLR pathway proteins as well as the expression of TLR2 and TLR4 will be performed by flow cytometry

BTK gene expressionwithin 24 hours of hospital admission (Day 0)

Measurements will be performed using the Quantitect Reverse Transcription Kit (Qiagen, Manchester, UK) for quantitative PCR (qPCR) on leucocytes.

Expression of X-linked miRNAs in leucocyteswithin 24 hours of hospital admission (Day 0)

Expression of X-linked miRNAs is measured by sequencing and qRT-PCR on leucocytes and/or plasma samples.

Expression of X-linked miRNAs in plasmawithin 24 hours of hospital admission (Day 0)

Expression of X-linked miRNAs is measured by sequencing and qRT-PCR on leucocytes and/or plasma samples.

Leukocyte populationDay 3

White blood cell count including neutrophils, monocytes, monocytes subtypes and lymphocytes. Only applicable for the sepsis sub-group

Total 17β-estradiolwithin 24 hours of hospital admission (Day 0)
Expression of the cell diapedesis receptor CD99 on lymphocyteswithin 24 hours of hospital admission (Day 0)

Measurements of cell diapedesis receptor CD99 on leukocytes will be performed by flow cytometry

Expression of TLR2 on PMNswithin 24 hours of hospital admission (Day 0)

Measurements of intracellular phosphorylated forms of TLR pathway proteins as well as the expression of TLR2 and TLR4 will be performed by flow cytometry

Expression of TLR4 on monocyteswithin 24 hours of hospital admission (Day 0)

Measurements of intracellular phosphorylated forms of TLR pathway proteins as well as the expression of TLR2 and TLR4 will be performed by flow cytometry

Expression of TLR4 on lymphocyteswithin 24 hours of hospital admission (Day 0)

Measurements of intracellular phosphorylated forms of TLR pathway proteins as well as the expression of TLR2 and TLR4 will be performed by flow cytometry

NEMO gene expressionwithin 24 hours of hospital admission (Day 0)

Measurements will be performed using the Quantitect Reverse Transcription Kit (Qiagen, Manchester, UK) for quantitative PCR (qPCR) on leucocytes.

Expression of TLR2 on lymphocyteswithin 24 hours of hospital admission (Day 0)

Measurements of intracellular phosphorylated forms of TLR pathway proteins as well as the expression of TLR2 and TLR4 will be performed by flow cytometry

Expression of TLR4 on PMNswithin 24 hours of hospital admission (Day 0)

Measurements of intracellular phosphorylated forms of TLR pathway proteins as well as the expression of TLR2 and TLR4 will be performed by flow cytometry

IRAK1 gene expressionwithin 24 hours of hospital admission (Day 0)

Measurements will be performed using the Quantitect Reverse Transcription Kit (Qiagen, Manchester, UK) for quantitative PCR (qPCR) on leucocytes.

CRPDay 3

Only applicable for the sepsis sub-group

Testosteronewithin 24 hours of hospital admission (Day 0)
pSOFA scoreDay 3

Only applicable for the sepsis sub-group. The pSOFA will be evaluated every 24 hours in order to compare laboratory and clinical data. The score will be based on the PaO2: FiO2 or SpO2: FiO2 ratio, the platelet count, the bilirubin level, the Mean Arterial Pressure (MAP), the Glasgow score and the creatinine level.

Microbiome analysisDuring subject hospitalisation

Trial Locations

Locations (1)

HUDERF

🇧🇪

Brussels, Belgium

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