To Develop an Algorithm for Predicting the Unfavorable Course of Sepsis in Children Based on a Comprehensive Assessment of Immunological, Biochemical and Molecular Genetic Markers

Study Overview
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Brief Summary
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Current study evaluates the relationship between cell immunity, biochemical and genetic markers in patients with sepsis in order to develop algorithm for predicting the course and outcome of severe bacterial infections.

Detailed Description
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To study the state of the immune system in patients with sepsis: the content of blood monocyte subpopulations (classical, intermediate and non-classical monocytes), HLA-DR expression on monocytes, CD64 expression on neutrophils, spontaneous and stimulated production of reactive oxygen species by blood neutrophils etc.; To study biochemical blood parameters and genetic polymorphism of immune response genes; To evaluate the relationship between genetic features of the patient's immune system, biochemical blood parameters, immune system cells and the severity of the pathological process. To develop an algorithm to prevent the development of sepsis in children.

Official Title
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To Develop an Algorithm for Predicting the Unfavorable Course of Sepsis in Children Based on a Comprehensive Assessment of Immunological, Biochemical and Molecular Genetic Markers

Conditions
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Sepsis

Intervention / Treatment
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* Diagnostic Test: flow cytometry
* Diagnostic Test: ELISA
* Diagnostic Test: RFLP


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: community-acquired pneumonia, severe course (according to IDSA / ATs criteria) and / or sepsis with different localization of the primary focus (SOFA> 2); availability of written informed consent of the patient's legal representatives for the study. Exclusion Criteria: acute and chronic heart failure; renal failure; acute and chronic liver failure; acute and chronic leukemia, severe anemia; HIV infection; immunosuppressive therapy; malignant neoplasms

Ages Eligible for Study
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Minimum Age: 1 Month
Maximum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| control<br>healthy children | Diagnostic Test: flow cytometry<br>* Determination of monocytes subpopulations (classical monocytes (CD14++CD16-, intermediate monocytes (CD14++CD16+ and non-classical monocytes (CD14+/-CD16++) in the blood of patients, evaluation of the HLA-DR expression on monocytes, evaluation of CD64 expression on neutrophils<br>Diagnostic Test: ELISA<br>* estimation of the TNF-α, LPS, IL-2, IL-4, IL-6, IL-8, IL-10 levels in the serum<br>Diagnostic Test: RFLP<br>* assessment of polymorphism TNF-α, LTA, IL-4, IL-6, IL-8, IL-10, TLR-2, TLR-4 genes with restriction fragment length polymorphism technique<br>|
| sepsis-1<br>children with sepsis on the 1 day | Diagnostic Test: flow cytometry<br>* Determination of monocytes subpopulations (classical monocytes (CD14++CD16-, intermediate monocytes (CD14++CD16+ and non-classical monocytes (CD14+/-CD16++) in the blood of patients, evaluation of the HLA-DR expression on monocytes, evaluation of CD64 expression on neutrophils<br>Diagnostic Test: ELISA<br>* estimation of the TNF-α, LPS, IL-2, IL-4, IL-6, IL-8, IL-10 levels in the serum<br>Diagnostic Test: RFLP<br>* assessment of polymorphism TNF-α, LTA, IL-4, IL-6, IL-8, IL-10, TLR-2, TLR-4 genes with restriction fragment length polymorphism technique<br>|
| sepsis-7<br>children with sepsis on the 7 day | Diagnostic Test: flow cytometry<br>* Determination of monocytes subpopulations (classical monocytes (CD14++CD16-, intermediate monocytes (CD14++CD16+ and non-classical monocytes (CD14+/-CD16++) in the blood of patients, evaluation of the HLA-DR expression on monocytes, evaluation of CD64 expression on neutrophils<br>Diagnostic Test: ELISA<br>* estimation of the TNF-α, LPS, IL-2, IL-4, IL-6, IL-8, IL-10 levels in the serum<br>Diagnostic Test: RFLP<br>* assessment of polymorphism TNF-α, LTA, IL-4, IL-6, IL-8, IL-10, TLR-2, TLR-4 genes with restriction fragment length polymorphism technique<br>|
| severe bacterial infection<br>children with severe bacterial infection | Diagnostic Test: flow cytometry<br>* Determination of monocytes subpopulations (classical monocytes (CD14++CD16-, intermediate monocytes (CD14++CD16+ and non-classical monocytes (CD14+/-CD16++) in the blood of patients, evaluation of the HLA-DR expression on monocytes, evaluation of CD64 expression on neutrophils<br>Diagnostic Test: ELISA<br>* estimation of the TNF-α, LPS, IL-2, IL-4, IL-6, IL-8, IL-10 levels in the serum<br>Diagnostic Test: RFLP<br>* assessment of polymorphism TNF-α, LTA, IL-4, IL-6, IL-8, IL-10, TLR-2, TLR-4 genes with restriction fragment length polymorphism technique<br>|
| severe viral infection<br>children with severe viral infection | Diagnostic Test: flow cytometry<br>* Determination of monocytes subpopulations (classical monocytes (CD14++CD16-, intermediate monocytes (CD14++CD16+ and non-classical monocytes (CD14+/-CD16++) in the blood of patients, evaluation of the HLA-DR expression on monocytes, evaluation of CD64 expression on neutrophils<br>Diagnostic Test: ELISA<br>* estimation of the TNF-α, LPS, IL-2, IL-4, IL-6, IL-8, IL-10 levels in the serum<br>Diagnostic Test: RFLP<br>* assessment of polymorphism TNF-α, LTA, IL-4, IL-6, IL-8, IL-10, TLR-2, TLR-4 genes with restriction fragment length polymorphism technique<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| monocytes | proportion of classical monocytes (CD14++CD16-), intermediate monocytes (CD14++CD16+) and non-classical monocytes (CD14+/-CD16++); HLA-DR level of expression | 3 years | 
| neutrophils | percentage of CD64+ neutrophils and CD64 level of expression | 3 years | 
| cytokines levels in the serum | TNF-α, LPS, IL-2, IL-4, IL-6, IL-8, IL-10 levels in the serum | 3 years | 
| genes polymorphism | TNF-α, LTA, IL-4, IL-6, IL-8, IL-10, TLR-2, TLR-4 genes polymorphism | 3 years | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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monocytes, neutrophils, genetic polymorphism