Potential Role of Inflammasome NLRP3 And IL-1B Gene Expression in COVID-19 Patients: Impact of Ferretin and D -Dimer.

Study Overview
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Brief Summary
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2.1 Study the role of NLRP3 inflammasome in COVID-19 patients. 2.2 Study the gene expression of NLRP3 and IL-1β in blood samples of COVID-19 patients and compare to apparently healthy subjects. 2.3 Correlation between NLRP3, IL-1β, IL-6 and severity of the disease. 2.4 Impact of ferritin and D-dimer on inflammasome componnets NLRP3, IL-1β IL-6 .

Detailed Description
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Coronavirus Disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a viral infection that results in respiratory disease, which can evolve into multiorgan failure (MOF), leading to death (Aida A Abdelmaksoud et al., 2021). The first cases of COVID-19 were detected in Wuhan, China, in 2019. Since then, the illness has spread rapidly around the country and the world reaching a pandemic level (Rocklöv J et al., 2020). Cases have been reported in more than 180 countries to the World Health Organization (WHO), including more than two million deaths (WHO, 2021). Several biochemical alterations have been described in COVID-19 patients as lymphopenia, thrombocytopenia and increased levels of C-reactive protein. The hallmark of severe COVID-19 is the cytokine storm accompanied by a hyperinflammatory response in the host due to the release of large amounts of pro-inflammatory cytokines IL-1β, IL-6, IL-2, IL-7, TNF-α, interferon-γ (IFN)-γ, CRP, procalcitonin (PCT), lactate dehydrogenase (LDH), erythrocyte sedimentation rate (ESR) and ferritin (Marcello Ciaccio and Agnello, 2020; Shah A., 2020). The inflammasome is a multiprotein complex, known for its role in the innate immune response against viral diseases and a regulator of processing of the pro-inflammatory cytokines interleukin (IL)-1β and IL-18. Inflammasome contains a NOD-like receptor (NLR) sensor protein and pyrin domain-containing 3 , after which, NLRP3 Inflammasome was named (de Rivero Vaccari et al., 2020). The presence of inflammasome-derived products such as IL-1β, IL-18, and LDH in patients' sera suggests inflammasome engagement (Chen G et al., 2020). Increased IL-6, IL-10, and IL-4 were found in patients when compared with controls, indicating that SARS-CoV-2 infects human monocytes and triggers NLRP3 inflammasome activation. However, the definitive demonstration of NRLP3 inflammasome participation is still required because these products can be produced via inflammasome-independent pathways (Rodrigues et al., 2021). But, the possible contribution of NLRP3 inflammasome activation and their ability to induce IL-1β production is still not clear . The pronounced inflammatory characteristics of COVID-19 prompted us to investigate the expression of the NRLP3 inflammasome and its relation to IL-1B in disease development and severity and to determine the best anti-inflammatory will be used in COVID-19.

Official Title
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Potential Role of Inflammasome NLRP3 And IL-1B Gene Expression In COVID-19 Patients: Impact of Ferretin and D -Dimer.

Conditions
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Inflammasome NLRP3 And IL-1B Gene Expression in COVID-19 Patients

Intervention / Treatment
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* Genetic: Quantitative Real Time Polymerase chain reaction (qRT-PCR)
* Diagnostic Test: ELISA and colorimeter


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Adult Patient aged 18years and over presented to Assiut University Hospital Suspected of covid19 because of the presence of symptoms suggestive of pneumonia (fever>38) and at least one of the following symptoms ;cough, dyspnea, tachpnea or hypoxia having RT-PCR (confirmed positive PCR) Chest CT within 5 days of initial PCR Exclusion Criteria: Cases less than 18 years old Cancer patients (affect result of study as they have disrubted level of these biomarker)

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 90 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 |
| --- | --- |
| healthy<br>as a control group | Genetic: Quantitative Real Time Polymerase chain reaction (qRT-PCR)<br>* to study gene expression of NLRP3 inflammasome and IL-1B in blood<br>* Other names: qRT-PCR;Diagnostic Test: ELISA and colorimeter<br>* IL-6 and total LDH<br>|
| mild to moderate covid-19<br>mild-moderate symptoms | Genetic: Quantitative Real Time Polymerase chain reaction (qRT-PCR)<br>* to study gene expression of NLRP3 inflammasome and IL-1B in blood<br>* Other names: qRT-PCR;Diagnostic Test: ELISA and colorimeter<br>* IL-6 and total LDH<br>|
| sever covid-19.<br>severe symptoms | Genetic: Quantitative Real Time Polymerase chain reaction (qRT-PCR)<br>* to study gene expression of NLRP3 inflammasome and IL-1B in blood<br>* Other names: qRT-PCR;Diagnostic Test: ELISA and colorimeter<br>* IL-6 and total LDH<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| gene expression of NLRP3, IL-1β and level IL-6 | Fold change increased in gene expression of NLRP3, IL-1β and level IL-6 and severity of the disease | 3 years | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| ferritin and D-dimer on NLRP3, IL-1β and IL-6 | The relation between level of ferritin and D-dimer on NLRP3, IL-1β and IL-6 | 3 years |