Ignite Creation Date:
2024-05-06 @ 8:00 PM
Last Modification Date:
2024-10-26 @ 3:18 PM
Study NCT ID:
NCT06217471
Status:
COMPLETED
Last Update Posted:
2024-01-22
First Post:
2023-12-27
Brief Title:
Influenza A VIRus and Destabilization of Atherosclerotic Carotid Plaques
Sponsor:
IRCCS Policlinico S Donato
Organization:
IRCCS Policlinico S Donato
Study Overview
Official Title:
Role of Influenza A VIRus in the Biomolecular Pathogenesis of the Destabilization of Atherosclerotic Carotid Plaques
Status:
COMPLETED
Status Verified Date:
2024-01
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
VIRAL
Brief Summary:
Atherosclerosis is the main cause of cardiovascular diseases and is characterized by the accumulation of lipids and inflammatory cells such as macrophages and lymphocytes within the vessel wall of large and medium-sized arteries forming so-called plaques The underlying molecular mechanisms are not yet clearly understood In particular it is not yet clear what factors can cause the destabilization of atherosclerotic plaques thus making them more vulnerable and prone to triggering acute cardiovascular events Infectious agents have also been implicated in the pathogenesis of atherosclerosis Some of them would be able to spread from the infected tissue and migrate to endothelial cells promoting the secretion of inflammatory mediators and the oxidation of low-density lipoproteins LDL their accumulation in vascular cells and the formation of foam cells fundamental mechanisms especially in the formation of vulnerable plaques
Recently many studies have shown that the influenza virus can also play a role in the destabilization of atherosclerotic plaques However the role of influenza A virus IVA infection and related vaccination in the destabilization of atherosclerotic plaques is still controversial Furthermore the underlying molecular mechanisms are still a matter of investigation
Based on these data we hypothesized that IV A infection may promote the destabilization of atherosclerotic plaques through a chronic postinfection immune response This response would lead to systemic and local changes in the expression of pro-atherosclerotic cytokines and chemokines resulting in increased recruitment of monocyte macrophages and upregulation of the expression of scavenger receptors on the surface of macrophages with greater affinity for oxidized LDL CD36 and Lectins- Like-oxLDL-receptor 1
Recently many studies have shown that the influenza virus can also play a role in the destabilization of atherosclerotic plaques However the role of influenza A virus IVA infection and related vaccination in the destabilization of atherosclerotic plaques is still controversial Furthermore the underlying molecular mechanisms are still a matter of investigation
Based on these data we hypothesized that IV A infection may promote the destabilization of atherosclerotic plaques through a chronic postinfection immune response This response would lead to systemic and local changes in the expression of pro-atherosclerotic cytokines and chemokines resulting in increased recruitment of monocyte macrophages and upregulation of the expression of scavenger receptors on the surface of macrophages with greater affinity for oxidized LDL CD36 and Lectins- Like-oxLDL-receptor 1
Detailed Description:
The following data will be collected anonymously from the medical records
personal details age sex
comorbidity presence of cardiovascular risk factors arterial hypertension smoking dyslipidemia ischemic heart disease COPD diabetes mellitus chronic renal failure history of cerebrovascular ischemic events
history of flu vaccination
clinical weight height As per clinical practice patients will undergo carotid endarterectomy surgery Upon entry into the operating room patients will be taken 4 ml of blood in 4 tubes with a specific anticoagulant EDTA from a peripheral venous access already positioned for the surgical procedure regardless of the study These blood samples will be stored and processed appropriately for individual analyses
Samples of atherosclerotic plaque removed during surgery will also be stored appropriately for subsequent investigations
The data collected will be entered into the dedicated e-CRF and appropriately analyzed
Use of blood samples Peripheral blood mononuclear cells PBMC will be separated and isolated from the venous blood sample using a Percoll gradient These cells will be appropriately stimulated with influenza A virus antigens using specific commercial kits This process will allow the evaluation of immune reactivity to the virus both in the group of patients with vulnerable carotid plaque group A and in those with non-vulnerable plaque group B On the serum of the same samples the quantitative expression of some cytokineschemokines involved in inflammation and the atherosclerotic process IL-6 IL-1β TNF-α IFN-γ MCP-1 will be evaluated using the ELISA method
The appropriate antibody evaluation will also be performed to test for any acute IV A infections
Use of vascular samples Histology investigations will be carried out on atherosclerotic plaque samples which will allow determining the vulnerability or otherwise of the plaque in such a way as to define the two groups of patients A vulnerable plaque B non-vulnerable plaque to be analysed and compare
Western-Blot assays and immunohistochemical analyzes will also be performed on the same samples to evaluate the quantitative and qualitative expression of some molecules V-CAM I-CAM E-Selectins and receptors that identify scavenger macrophage cells CD36 and Lectins -Like Ox-LDL-Receptor 1
Finally the same samples will be subjected to ELISA analysis for the quantitative research of the same cytokineschemokines evaluated on the blood sample via RT-PCR The possible presence of viral RNA will be evaluated on the atherosclerotic plaques by one step RT-PCR as well as the T-specific cell reactivity to the IV A virus after the preparation of appropriate cell cultures which will be stimulated with the same commercial kit used to stimulate the PMBC of peripheral blood according to the method described by Keller et al Collaterally the possible presence of viral or bacterial DNA in the plaques will be checked In particular the presence of DNA from Human Polyomaviruses and Human Herpesviruses herpes simplex virus 7 and 2 varicella zoster virus Epstein-Barr virus human cytomegalovirus human herpesvirus 6 and Chlamydia will be analyzed through Real Time PCR
Evaluation of specific T-cell reactivity to virus IV A Taking as reference the method described by Keller et al the stimulation index SI will be evaluated both on the blood sample and on the atherosclerotic plaque as the average count per minute cpm of cultures with the virus divided by the mean cpm of parallel cultures without viruses
Subsequently the reactivity will be calculated based on the ratio between the SI of the plaque T cells and the SI of the peripheral blood T cells for each patient defining responsive patients if the ratio is 2 patients with SI of the T cells T of the plaque significantly higher than those of the peripheral blood and not responsive if the ratio is 2 in the latter case patients with SI of the T cells of the peripheral blood greater than those of the plaque or with SI of the T cells Plaque T not significantly higher than those of peripheral blood
personal details age sex
comorbidity presence of cardiovascular risk factors arterial hypertension smoking dyslipidemia ischemic heart disease COPD diabetes mellitus chronic renal failure history of cerebrovascular ischemic events
history of flu vaccination
clinical weight height As per clinical practice patients will undergo carotid endarterectomy surgery Upon entry into the operating room patients will be taken 4 ml of blood in 4 tubes with a specific anticoagulant EDTA from a peripheral venous access already positioned for the surgical procedure regardless of the study These blood samples will be stored and processed appropriately for individual analyses
Samples of atherosclerotic plaque removed during surgery will also be stored appropriately for subsequent investigations
The data collected will be entered into the dedicated e-CRF and appropriately analyzed
Use of blood samples Peripheral blood mononuclear cells PBMC will be separated and isolated from the venous blood sample using a Percoll gradient These cells will be appropriately stimulated with influenza A virus antigens using specific commercial kits This process will allow the evaluation of immune reactivity to the virus both in the group of patients with vulnerable carotid plaque group A and in those with non-vulnerable plaque group B On the serum of the same samples the quantitative expression of some cytokineschemokines involved in inflammation and the atherosclerotic process IL-6 IL-1β TNF-α IFN-γ MCP-1 will be evaluated using the ELISA method
The appropriate antibody evaluation will also be performed to test for any acute IV A infections
Use of vascular samples Histology investigations will be carried out on atherosclerotic plaque samples which will allow determining the vulnerability or otherwise of the plaque in such a way as to define the two groups of patients A vulnerable plaque B non-vulnerable plaque to be analysed and compare
Western-Blot assays and immunohistochemical analyzes will also be performed on the same samples to evaluate the quantitative and qualitative expression of some molecules V-CAM I-CAM E-Selectins and receptors that identify scavenger macrophage cells CD36 and Lectins -Like Ox-LDL-Receptor 1
Finally the same samples will be subjected to ELISA analysis for the quantitative research of the same cytokineschemokines evaluated on the blood sample via RT-PCR The possible presence of viral RNA will be evaluated on the atherosclerotic plaques by one step RT-PCR as well as the T-specific cell reactivity to the IV A virus after the preparation of appropriate cell cultures which will be stimulated with the same commercial kit used to stimulate the PMBC of peripheral blood according to the method described by Keller et al Collaterally the possible presence of viral or bacterial DNA in the plaques will be checked In particular the presence of DNA from Human Polyomaviruses and Human Herpesviruses herpes simplex virus 7 and 2 varicella zoster virus Epstein-Barr virus human cytomegalovirus human herpesvirus 6 and Chlamydia will be analyzed through Real Time PCR
Evaluation of specific T-cell reactivity to virus IV A Taking as reference the method described by Keller et al the stimulation index SI will be evaluated both on the blood sample and on the atherosclerotic plaque as the average count per minute cpm of cultures with the virus divided by the mean cpm of parallel cultures without viruses
Subsequently the reactivity will be calculated based on the ratio between the SI of the plaque T cells and the SI of the peripheral blood T cells for each patient defining responsive patients if the ratio is 2 patients with SI of the T cells T of the plaque significantly higher than those of the peripheral blood and not responsive if the ratio is 2 in the latter case patients with SI of the T cells of the peripheral blood greater than those of the plaque or with SI of the T cells Plaque T not significantly higher than those of peripheral blood
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None