Ignite Creation Date:
2024-10-26 @ 3:41 PM
Last Modification Date:
2024-10-26 @ 3:41 PM
Study NCT ID:
NCT06612554
Status:
NOT_YET_RECRUITING
Last Update Posted:
None
First Post:
2024-09-15
Brief Title:
Zn Supplementation in HIV Immunological Non Responders
Sponsor:
None
Organization:
None
Study Overview
Official Title:
Role of Zinc Supplementation in Immunological Non-responders HIV Individuals Exploring Pathways for Persistent Inflammation
Status:
NOT_YET_RECRUITING
Status Verified Date:
2024-09
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
No
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
VIHZn
Brief Summary:
Zinc is an essential micronutrient crucial for the normal functioning of the human immune system Zinc deficiency impairs immune function and increases infection risk especially in vulnerable populations like people living with HIV This project aims to study the role of zinc supplementation in improving immune response in HIV-infected individuals who are Immunological Non-Responders INRs-people who have not restored Cluster of differentiation 4 CD4 T-cell counts despite receiving antiretroviral therapy ART INRs face a higher risk of opportunistic infections non-HIV comorbidities due to high inflammation and generally have a poorer prognosis Zinc supports immune function by aiding in immune cell development cytokine production and maintaining mucosal barrier integrity
Several studies have investigated zinc supplementation in HIV-infected individuals showing significant increases in CD4 counts and a reduction in opportunistic infections However the doses used vary and the results are sometimes contradictory Our objective is to study whether zinc supplementation in INRs improves immune function specifically by increasing CD4 counts decreasing inflammation and affecting other immune parameters
This project involves a clinical trial where INRs will be randomly assigned to receive 75mg of elemental zinc daily in the form of 3 zinc acetate tablets along with their usual treatment or to continue their treatment without zinc supplements We will assess whether zinc supplementation increases CD4 lymphocytes reduces inflammation in INRs and induces immune system changes We will also investigate immune system functionality by measuring the presence of the Torque Teno Virus TTV a harmless virus and see how zinc supplementation impacts its control by monitoring viral load changes
Recent research by our group has demonstrated that zinc has both antiviral and anti-inflammatory effects Zinc supplementation is generally safe and well-tolerated with few adverse effects Zinc is available in various forms including gluconate acetate and sulfate Although the recommended daily dose is 40mg studies have shown that doses exceeding 80mg have been safely administered for over 10 years without side effects
We have selected a 75mg daily dose of elemental zinc for several reasons Studies that showed no effect used lower doses 15-20mgday and our research found that 75mgday improved outcomes in acute viral infections like Severe Acute Respiratory Syndrome-Coronavirus-2 SARS-CoV2 We believe previous failures to show zincs efficacy were due to insufficient dosing After extensive literature review we concluded that 75mg daily is safe and likely to produce the desired effects
Our goal is to demonstrate the efficacy of zinc as an immunomodulatory agent If successful this could be a simple and cost-effective way to improve the quality of life for many people We would recommend its widespread use under medical supervision particularly at the doses being studied
Several studies have investigated zinc supplementation in HIV-infected individuals showing significant increases in CD4 counts and a reduction in opportunistic infections However the doses used vary and the results are sometimes contradictory Our objective is to study whether zinc supplementation in INRs improves immune function specifically by increasing CD4 counts decreasing inflammation and affecting other immune parameters
This project involves a clinical trial where INRs will be randomly assigned to receive 75mg of elemental zinc daily in the form of 3 zinc acetate tablets along with their usual treatment or to continue their treatment without zinc supplements We will assess whether zinc supplementation increases CD4 lymphocytes reduces inflammation in INRs and induces immune system changes We will also investigate immune system functionality by measuring the presence of the Torque Teno Virus TTV a harmless virus and see how zinc supplementation impacts its control by monitoring viral load changes
Recent research by our group has demonstrated that zinc has both antiviral and anti-inflammatory effects Zinc supplementation is generally safe and well-tolerated with few adverse effects Zinc is available in various forms including gluconate acetate and sulfate Although the recommended daily dose is 40mg studies have shown that doses exceeding 80mg have been safely administered for over 10 years without side effects
We have selected a 75mg daily dose of elemental zinc for several reasons Studies that showed no effect used lower doses 15-20mgday and our research found that 75mgday improved outcomes in acute viral infections like Severe Acute Respiratory Syndrome-Coronavirus-2 SARS-CoV2 We believe previous failures to show zincs efficacy were due to insufficient dosing After extensive literature review we concluded that 75mg daily is safe and likely to produce the desired effects
Our goal is to demonstrate the efficacy of zinc as an immunomodulatory agent If successful this could be a simple and cost-effective way to improve the quality of life for many people We would recommend its widespread use under medical supervision particularly at the doses being studied
Detailed Description:
Studies of people living with HIV show that although antiretroviral treatments ART decreases inflammation a number of inflammatory markers remain persistently elevated and this chronic inflammation can play an important role in cardiovascular disease CVD Moreover some individuals with HIV initiating ART do not recover Cluster of differentiation 4 CD4 T-cells count as expected and are described as Immunological Non-Responders INRs INRs present with severely altered immunological functions including malfunction and diminished production of cells within lymphopoietic tissue perturbed frequencies of immune regulators such as regulatory T cells and Th17 cells and increased immune activation immunosenescence and apoptosis Importantly INRs have an increased risk of morbidity and mortality compared to HIV-infected patients with an optimal immune reconstitution Our group has demonstrated how this group responds poorly to messenger RNA mRNA vaccination against Severe Acute Respiratory Syndrome-Coronavirus-2 SARS-CoV2 showing an impaired immune function among these individuals
However the definition of Immunological Non-Responders INRs suffers from lack of consensus impeding the comparison of findings There seems to be agreement that an adequate immune response to antiretroviral treatment should include a CD4 cells count more than 500 cellsμL mainly because HIV-infected patients with this level of immune restoration have a morbidity and mortality rate approaching or comparable to those of HIV negative individuals Patients with CD4 cell counts 500 cellsμL are consequently classified as inadequate responders included in the INRs group And although they are a group poorly described their morbidity and mortality seem to be more similar to INRs than to adequate responders
The underlying mechanism that explains INRs phenomena is not well understood Several hypotheses have been raised as an older age a long duration of HIV infection prior to antiretroviral treatment co-infection with hepatitis C and a low CD4 nadir predispose to immunological nonresponse The CD4 nadir specifically appears to be critical for the recovery of CD4 cells However none of these factors provide a full explanation for the lack of immune reconstitution in INRs Probably this is the consequence of several factors and its interaction Interestingly low plasma zinc levels and inadequate zinc intake were found in up 50 of participants in several cohorts of HIV infected adults and were independently associated with faster HIV disease progression and increased mortality Zinc deficiency reduces generation of T-cells depresses humoral and cell-mediated immunity leads to lymphopenia and thymic atrophy and increases the frequency and number of infections Thus the zinc status could also have a role in the mechanisms involved in INRs
Zinc in viral infection Zinc is an essential trace element for both host and pathogens In order to grow pathogens require zinc Thus zinc is a structural cofactor of viral proteins and certain viruses have developed strategies to alter cellular zinc homeostasis on its benefit On the other hand there is extensive evidence that zinc can prevent viral replication and lower cellular invasion eg Rhinovirus Hepatitis CSARS-CoV2
Remarkably our team has shown that low zinc levels favor SARS-CoV2 expansion These results might indicate either a direct anti-viral action against SARS-CoV2 or that the virus benefits from stress conditions caused by low levels of cellular zinc Zinc supplementation has been shown to be beneficial when administering to Hepatitis B and Hepatitis C infected patients Moreover zinc-based nutritional immunity reduces diarrheal episodes and respiratory tract diseases Remarkably zinc supplementation for the common cold caused by rhino- and coronaviruses prevents it and reduces its duration if the administration starts within 24h after the onset of symptoms Thus zinc supplementation has been proven beneficial against some viral infections However whether this was caused by improved local immune response or viral inhibition remains uncertain
Zinc as immunomodulator Zinc deficiency ZD is known to be associated with proinflammatory responses at infection showing higher reactive oxygen species ROS production and inflammatory markers An imbalance in cytokine production by cells of both innate and adaptive immunity has also been reported
Concretely ZD is associated with higher levels of Interleukin-6 IL-6 Interleukin-1beta IL-1beta and Tumor Necrosis Factor-alpha TNF-a In this scenario zinc supplementation has been shown to decrease the incidence of infection inflammatory cytokines including IL-6 and oxidative stress markers in elderly individuals Moreover zinc supplementation has been shown to balance immune responses
Nutritional zinc status and IL-6 are highly interconnected Thus IL-6 can reduce zinc bioavailability promoting internalization in hepatocytes and the expression of zinc chelators On the other hand zinc decreases IL-6 production via inhibition of Signal Transducer and Activator of Transcription-3 STAT-3 pathway Our study with Coronavirus Disease-19 COVID-19 patients showed a negative correlation between serum zinc levels and IL-6 production
Zinc in T lymphocytes activation and exhaustion Zinc is known to affect T lymphocyte maturation differentiation and cytokine production Activated T cells are known to increase zinc content On the one hand zinc has been shown to be required for the correct T-cell Receptor TCR signaling by modulating Lck and Zap- 70 activity Our collaborators Vicente et al have demonstrated that zinc positively potentiates the three main signaling pathways Nuclear Factor-kB NF-kb and Nuclear Factor of Activated T-cells-1 The knock-down of Zip6 a zinc transporter up regulated early during cell activation alters zinc entry and dramatically impairs activation of Jurkat T cells Moreover recent studies on Cluster of differentiation 8 CD8 T-cell dysfunction during cancer and chronic infections have started to elucidate the transcriptional pathways involved in this phenomenon and zinc homeostasis has emerged again as a key element Thus metallothioneins 1 and 2 levels and zinc dependent transcription factors are differentially expressed in exhausted CD8 cells Nonetheless there are no studies investigating whether zinc supplementationchelation might directly affect the CD8 dysfunctional profile and how these learnings could be translated into novel therapeutic approaches for these diseases including HIV infection
Zinc in B lymphocyte function It has been shown that zinc transporters Zip7 and Zip10 are essential in B cell function B cells activation and plasma cell differentiation depends on zinc signaling Nutritional zinc status is an important factor determining antibody production given its action on both B-cells and CD4 T lymphocytes Thus ZD has been shown in mouse models to lower humoral immune response
Preliminary Results Our group has been working in the immunomodulatory effect of zinc in viral infections and in the immune response since 2020 Also we are actively working in the immune response in people with HIV PWH INRs We have several publications and an active research line putting these two research lines together
We previously carried out a study analyzing the association between zinc plasma levels at hospital admission and COVID-19 prognosis during the first wave of the pandemics We found a negative correlation between the zinc levels and IL-6 as well as with mortality We also found that zinc acted on viral replication in vero cell cultures We have recently conducted a clinical trial NCT05778383 whose preliminary results have been reported in CROI2023 542 In this trial we observed that high-dose Zinc supplementation in the acute phase of SARS-coV-2 infection is associated with a better prognosis of the disease We also shown an abnormal immune response to vaccination among INRs Therefore we have demonstrated in a viral infection context that zinc has an antiviral and anti-inflammatory effect which may have a benefit in people living with HIV especially in the INR situation
Hypothesis
This study has several hypotheses
1 Relative low zinc levels are frequent in our cohort of HIV patients and have a role in worse immune response and in higher inflammatory response
2 We hypothesize that zinc has an impact in the immune system restoring the lymphocyte count in INRs HIV individuals and improving immune function and inflammation associated with chronic viral infection
Therefore zinc supplementation in physiological concentrations and recommendations has a beneficial impact in immune function reducing immune activation and reducing chronic inflammation in INRs HIV individuals
3 Zinc supplementation protects gut mucosa of cytopathic effect of virus restoring partially the gut permeability induced by HIV
4 Zinc supplementation improves oxidative stress and mitochondrial function
Objectives
The main objective is to study the clinical and immune benefits of 16 weeks of supplementation with zinc acetate in HIV immunological non-responders individuals To address this objective a Randomized Clinical Trial RCT will be designed
Aim 1 To design an double blinded randomized multicentric 2-arm study The following parameters will be measured at baseline and at week-16 in zinc-supplemented individuals vs controls
Primary Change in CD4 T-cell count
Secondary Changes in Quality of life changes through EuroQol-5D-3L CD4 CD8 T-cell activation T-cell subsets Torque Teno Virus replication IL-6 C-RP Transcriptomics ROS
Aim 2 To measure persistence in changes quality of life immune function immune activation inflammatory biomarkers and mitochondrial function and ROS at week 24 respect to week-16 and baseline In terms of Changes in Quality of life changes through EuroQol-5D-3L CD4 CD8 T-cell activation T-cell subsets Torque Teno Virus replication IL-6 C-RP Transcriptomics ROS
The methodology is divided into tasks
Task 1 Randomized Clinical Trial An open label randomized multicentric 2-arm study will be designed Participants will be prospectively enrolled at two sites Hospital del Mar and Hospital Parc Taulí In the Zinc Supplementation group Zn participants will received the Standard of Care SoC and separated 12h just in case the ART is based in Integrase Inhibitors INSTI will be supplemented during 16 weeks with 75mg of elemental Zn 3 tablets 83mg Zn acetate 249mg of Zinc Acetate Zinc-NM Laboratorios Cantabria Spanish National Code 1562524 We have selected 75mg of elemental Znd as oral supplementation because it has been used safely in non-HIV studies with 1 year duration at this concentration with no serious adverse effects 15 End of Treatment EoT will be at 16 weeks but a follow up up to 24-weeks will be done to assess the persistence of the effects of Zn
The study involves multiple tasks across different visits and sample collections aiming to assess the effects of the intervention on immune response inflammation and oxidative stress in participants
Task 11 Visits and Sample Collection
Participants will undergo three visits
Visit 1 ScreeningRandomizationBaseline In-person visit where randomization is performed using REDCap After informed consent medical history is recorded vital signs are taken and blood samples are collected for various tests including CD4 T-cell count HIV viral load C-reactive protein CRP ferritin D-dimer vitamin D and zinc Participants also complete the EuroQol-5D-3L quality of life questionnaire Blood samples are collected in serum ethylenediaminetetraacetic acid EDTA and RNA tubes for peripheral blood mononuclear cells PBMCs
Visit 2 Week 16 Similar assessments as Visit 1 are performed with the addition of tracking adverse events AEs and concomitant therapies
Visit 3 Week 24 The same tests are repeated to extend the study with sample collection focusing on the same biomarkers as in prior visits
Task 2 T-cell Subset Assessment PBMCs from the enrolled participants will be isolated using density gradient centrifugation Samples are preserved at -80C and sent to IMIM for processing Flow cytometry will be used to analyze T-cell subsets including markers like CD45 CD4 CD8 FOXP3 and PD-1 with data analyzed using FlowJo software
Task 3 Multiplex ELISA for Inflammatory Biomarkers At baseline Week 16 and Week 24 fasting blood sera will be collected to measure inflammatory markers such as sCD14 sCD163 CRP sTNFR-I sTNFR-II and gut integrity markers like IFABP and Lipopolysaccharide-binding protein LBP using ELISA
Task 4 Torque Teno Virus TTV Viral Load TTV viral load will be quantified using the TTV R-GENE kit with DNA extracted and analyzed via TaqMan real-time Polymerase Chain Reaction rt-PCR at baseline Week 16 and Week 24
Task 5 ROS and Mitochondrial Function Reactive Oxygen Species ROS are chemically reactive molecules containing oxygen such as free radicals and peroxides They are byproducts of cellular metabolism and play essential roles in cell signaling and homeostasis However excessive ROS can cause oxidative stress damaging DNA proteins and lipids which can lead to various diseases including cardiovascular diseases neurodegeneration and cancer
To assess ROS levels indirect markers such as glutathione GSHGSSG ratio 4-hydroxynonenal 4-HNE and malondialdehyde MDA are commonly measured Glutathione helps neutralize ROS with its GSHGSSG ratio indicating oxidative stress 4-HNE and MDA are byproducts of lipid peroxidation providing insights into cellular membrane damage due to ROS These markers are typically measured using techniques like enzymatic assays ELISA and TBARS assay respectively Understanding ROS levels and their effects is critical for studying oxidative stress and related diseases
Task 6 Transcriptomic Analysis RNA will be extracted from blood samples at baseline and Week 16 for transcriptomic analysis using expression microarrays Bioinformatics analysis will be conducted at the MARGenomics platform to study gene expression changes related to the intervention
B-Secondary outcomes
1-Change in the transcriptome profile between baseline and week 16 respect to week 24 in whole blood samples
C-Safety endpoints
-Incidence of adverse events AEs
However the definition of Immunological Non-Responders INRs suffers from lack of consensus impeding the comparison of findings There seems to be agreement that an adequate immune response to antiretroviral treatment should include a CD4 cells count more than 500 cellsμL mainly because HIV-infected patients with this level of immune restoration have a morbidity and mortality rate approaching or comparable to those of HIV negative individuals Patients with CD4 cell counts 500 cellsμL are consequently classified as inadequate responders included in the INRs group And although they are a group poorly described their morbidity and mortality seem to be more similar to INRs than to adequate responders
The underlying mechanism that explains INRs phenomena is not well understood Several hypotheses have been raised as an older age a long duration of HIV infection prior to antiretroviral treatment co-infection with hepatitis C and a low CD4 nadir predispose to immunological nonresponse The CD4 nadir specifically appears to be critical for the recovery of CD4 cells However none of these factors provide a full explanation for the lack of immune reconstitution in INRs Probably this is the consequence of several factors and its interaction Interestingly low plasma zinc levels and inadequate zinc intake were found in up 50 of participants in several cohorts of HIV infected adults and were independently associated with faster HIV disease progression and increased mortality Zinc deficiency reduces generation of T-cells depresses humoral and cell-mediated immunity leads to lymphopenia and thymic atrophy and increases the frequency and number of infections Thus the zinc status could also have a role in the mechanisms involved in INRs
Zinc in viral infection Zinc is an essential trace element for both host and pathogens In order to grow pathogens require zinc Thus zinc is a structural cofactor of viral proteins and certain viruses have developed strategies to alter cellular zinc homeostasis on its benefit On the other hand there is extensive evidence that zinc can prevent viral replication and lower cellular invasion eg Rhinovirus Hepatitis CSARS-CoV2
Remarkably our team has shown that low zinc levels favor SARS-CoV2 expansion These results might indicate either a direct anti-viral action against SARS-CoV2 or that the virus benefits from stress conditions caused by low levels of cellular zinc Zinc supplementation has been shown to be beneficial when administering to Hepatitis B and Hepatitis C infected patients Moreover zinc-based nutritional immunity reduces diarrheal episodes and respiratory tract diseases Remarkably zinc supplementation for the common cold caused by rhino- and coronaviruses prevents it and reduces its duration if the administration starts within 24h after the onset of symptoms Thus zinc supplementation has been proven beneficial against some viral infections However whether this was caused by improved local immune response or viral inhibition remains uncertain
Zinc as immunomodulator Zinc deficiency ZD is known to be associated with proinflammatory responses at infection showing higher reactive oxygen species ROS production and inflammatory markers An imbalance in cytokine production by cells of both innate and adaptive immunity has also been reported
Concretely ZD is associated with higher levels of Interleukin-6 IL-6 Interleukin-1beta IL-1beta and Tumor Necrosis Factor-alpha TNF-a In this scenario zinc supplementation has been shown to decrease the incidence of infection inflammatory cytokines including IL-6 and oxidative stress markers in elderly individuals Moreover zinc supplementation has been shown to balance immune responses
Nutritional zinc status and IL-6 are highly interconnected Thus IL-6 can reduce zinc bioavailability promoting internalization in hepatocytes and the expression of zinc chelators On the other hand zinc decreases IL-6 production via inhibition of Signal Transducer and Activator of Transcription-3 STAT-3 pathway Our study with Coronavirus Disease-19 COVID-19 patients showed a negative correlation between serum zinc levels and IL-6 production
Zinc in T lymphocytes activation and exhaustion Zinc is known to affect T lymphocyte maturation differentiation and cytokine production Activated T cells are known to increase zinc content On the one hand zinc has been shown to be required for the correct T-cell Receptor TCR signaling by modulating Lck and Zap- 70 activity Our collaborators Vicente et al have demonstrated that zinc positively potentiates the three main signaling pathways Nuclear Factor-kB NF-kb and Nuclear Factor of Activated T-cells-1 The knock-down of Zip6 a zinc transporter up regulated early during cell activation alters zinc entry and dramatically impairs activation of Jurkat T cells Moreover recent studies on Cluster of differentiation 8 CD8 T-cell dysfunction during cancer and chronic infections have started to elucidate the transcriptional pathways involved in this phenomenon and zinc homeostasis has emerged again as a key element Thus metallothioneins 1 and 2 levels and zinc dependent transcription factors are differentially expressed in exhausted CD8 cells Nonetheless there are no studies investigating whether zinc supplementationchelation might directly affect the CD8 dysfunctional profile and how these learnings could be translated into novel therapeutic approaches for these diseases including HIV infection
Zinc in B lymphocyte function It has been shown that zinc transporters Zip7 and Zip10 are essential in B cell function B cells activation and plasma cell differentiation depends on zinc signaling Nutritional zinc status is an important factor determining antibody production given its action on both B-cells and CD4 T lymphocytes Thus ZD has been shown in mouse models to lower humoral immune response
Preliminary Results Our group has been working in the immunomodulatory effect of zinc in viral infections and in the immune response since 2020 Also we are actively working in the immune response in people with HIV PWH INRs We have several publications and an active research line putting these two research lines together
We previously carried out a study analyzing the association between zinc plasma levels at hospital admission and COVID-19 prognosis during the first wave of the pandemics We found a negative correlation between the zinc levels and IL-6 as well as with mortality We also found that zinc acted on viral replication in vero cell cultures We have recently conducted a clinical trial NCT05778383 whose preliminary results have been reported in CROI2023 542 In this trial we observed that high-dose Zinc supplementation in the acute phase of SARS-coV-2 infection is associated with a better prognosis of the disease We also shown an abnormal immune response to vaccination among INRs Therefore we have demonstrated in a viral infection context that zinc has an antiviral and anti-inflammatory effect which may have a benefit in people living with HIV especially in the INR situation
Hypothesis
This study has several hypotheses
1 Relative low zinc levels are frequent in our cohort of HIV patients and have a role in worse immune response and in higher inflammatory response
2 We hypothesize that zinc has an impact in the immune system restoring the lymphocyte count in INRs HIV individuals and improving immune function and inflammation associated with chronic viral infection
Therefore zinc supplementation in physiological concentrations and recommendations has a beneficial impact in immune function reducing immune activation and reducing chronic inflammation in INRs HIV individuals
3 Zinc supplementation protects gut mucosa of cytopathic effect of virus restoring partially the gut permeability induced by HIV
4 Zinc supplementation improves oxidative stress and mitochondrial function
Objectives
The main objective is to study the clinical and immune benefits of 16 weeks of supplementation with zinc acetate in HIV immunological non-responders individuals To address this objective a Randomized Clinical Trial RCT will be designed
Aim 1 To design an double blinded randomized multicentric 2-arm study The following parameters will be measured at baseline and at week-16 in zinc-supplemented individuals vs controls
Primary Change in CD4 T-cell count
Secondary Changes in Quality of life changes through EuroQol-5D-3L CD4 CD8 T-cell activation T-cell subsets Torque Teno Virus replication IL-6 C-RP Transcriptomics ROS
Aim 2 To measure persistence in changes quality of life immune function immune activation inflammatory biomarkers and mitochondrial function and ROS at week 24 respect to week-16 and baseline In terms of Changes in Quality of life changes through EuroQol-5D-3L CD4 CD8 T-cell activation T-cell subsets Torque Teno Virus replication IL-6 C-RP Transcriptomics ROS
The methodology is divided into tasks
Task 1 Randomized Clinical Trial An open label randomized multicentric 2-arm study will be designed Participants will be prospectively enrolled at two sites Hospital del Mar and Hospital Parc Taulí In the Zinc Supplementation group Zn participants will received the Standard of Care SoC and separated 12h just in case the ART is based in Integrase Inhibitors INSTI will be supplemented during 16 weeks with 75mg of elemental Zn 3 tablets 83mg Zn acetate 249mg of Zinc Acetate Zinc-NM Laboratorios Cantabria Spanish National Code 1562524 We have selected 75mg of elemental Znd as oral supplementation because it has been used safely in non-HIV studies with 1 year duration at this concentration with no serious adverse effects 15 End of Treatment EoT will be at 16 weeks but a follow up up to 24-weeks will be done to assess the persistence of the effects of Zn
The study involves multiple tasks across different visits and sample collections aiming to assess the effects of the intervention on immune response inflammation and oxidative stress in participants
Task 11 Visits and Sample Collection
Participants will undergo three visits
Visit 1 ScreeningRandomizationBaseline In-person visit where randomization is performed using REDCap After informed consent medical history is recorded vital signs are taken and blood samples are collected for various tests including CD4 T-cell count HIV viral load C-reactive protein CRP ferritin D-dimer vitamin D and zinc Participants also complete the EuroQol-5D-3L quality of life questionnaire Blood samples are collected in serum ethylenediaminetetraacetic acid EDTA and RNA tubes for peripheral blood mononuclear cells PBMCs
Visit 2 Week 16 Similar assessments as Visit 1 are performed with the addition of tracking adverse events AEs and concomitant therapies
Visit 3 Week 24 The same tests are repeated to extend the study with sample collection focusing on the same biomarkers as in prior visits
Task 2 T-cell Subset Assessment PBMCs from the enrolled participants will be isolated using density gradient centrifugation Samples are preserved at -80C and sent to IMIM for processing Flow cytometry will be used to analyze T-cell subsets including markers like CD45 CD4 CD8 FOXP3 and PD-1 with data analyzed using FlowJo software
Task 3 Multiplex ELISA for Inflammatory Biomarkers At baseline Week 16 and Week 24 fasting blood sera will be collected to measure inflammatory markers such as sCD14 sCD163 CRP sTNFR-I sTNFR-II and gut integrity markers like IFABP and Lipopolysaccharide-binding protein LBP using ELISA
Task 4 Torque Teno Virus TTV Viral Load TTV viral load will be quantified using the TTV R-GENE kit with DNA extracted and analyzed via TaqMan real-time Polymerase Chain Reaction rt-PCR at baseline Week 16 and Week 24
Task 5 ROS and Mitochondrial Function Reactive Oxygen Species ROS are chemically reactive molecules containing oxygen such as free radicals and peroxides They are byproducts of cellular metabolism and play essential roles in cell signaling and homeostasis However excessive ROS can cause oxidative stress damaging DNA proteins and lipids which can lead to various diseases including cardiovascular diseases neurodegeneration and cancer
To assess ROS levels indirect markers such as glutathione GSHGSSG ratio 4-hydroxynonenal 4-HNE and malondialdehyde MDA are commonly measured Glutathione helps neutralize ROS with its GSHGSSG ratio indicating oxidative stress 4-HNE and MDA are byproducts of lipid peroxidation providing insights into cellular membrane damage due to ROS These markers are typically measured using techniques like enzymatic assays ELISA and TBARS assay respectively Understanding ROS levels and their effects is critical for studying oxidative stress and related diseases
Task 6 Transcriptomic Analysis RNA will be extracted from blood samples at baseline and Week 16 for transcriptomic analysis using expression microarrays Bioinformatics analysis will be conducted at the MARGenomics platform to study gene expression changes related to the intervention
B-Secondary outcomes
1-Change in the transcriptome profile between baseline and week 16 respect to week 24 in whole blood samples
C-Safety endpoints
-Incidence of adverse events AEs
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
None
Is a FDA Regulated Device?:
None
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None