Primary Objective:Our primary objective is to generate in-depth immune profiles, which include phenotypic and functional analyses, as well as epigenetic and transcriptomic analyses, of several B cell, T cell, and monocyte subsets, which relate to…
ID
Source
Brief title
Condition
- Autoimmune disorders
- Respiratory tract infections
Synonym
Research involving
Sponsors and support
Intervention
Outcome measures
Primary outcome
Percentage of age-associated B cells (ABCs; CD21loCD11c+ DN B cells) in
peripheral blood mononuclear cells (PBMCs) from long-COVID patients, compared
to convalescent, phenotypic, and autoimmune controls.
Secondary outcome
Immunological profile
The quantification of ABCs as primary parameter will be part of the
immunological profiling of long-COVID patients. This will be accompanied by the
characterization and quantification of several other parameters of circulating
lymphoid (B and T cell) and myeloid (monocytes and dendritic cells)
populations. Frequencies as well as the activation status of different subsets
will be determined in PBMCs from both patients and controls. In addition, the
expression of several activation markers (including CD86, MHC-II, CD69, etc.)
and exhaustion markers (PD-1, CTLA-4, etc.) will be determined. Functional
experiments will be conducted on B and T cells to measure B and T cell receptor
(BCR and TCR, respectively) signaling responsiveness. In addition, the cytokine
production potential will be evaluated in lymphoid and myeloid subsets to
assess functionality. Differences in the potential of cytokine production in
specific cell subsets between groups will be translated to a systemic level, by
quantifying circulating cytokine concentrations in plasma/serum. Quantification
and phenotypic assessment of SARS-CoV-2-specific B and T cells will be
performed using in-house techniques developed at the Department of Viroscience
[26]. Cell populations of interest will be further investigated on a
transcriptional and epigenetic level, using RNA- and ATAC-seq, respectively.
Finally, immunological characteristics will be combined and correlated with
clinical parameters to form computationally-calculated immunological profiles.
Virological profile
A virological profile will be created based on several circulating virological
factors. For SARS-CoV-2, these include the quantification of circulating RNA
and spike protein in serum/plasma. For reactivation of latent viruses, these
include the quantification of viral load of EBV, CMV, and HHV6 in serum/plasma.
Concentrations and neutralization capacity of antibodies (IgM and IgG) against
both SARS-CoV-2 and other latent viruses (anti-EBV, anti-CMV, and anti-HHV6)
will be determined in serum/plasma. Based on previous findings, SARS-CoV-2
protein will also be detected and quantified in circulating monocyte
subpopulations. Finally, these virological characteristics will be combined and
correlated with clinical parameters to form a computationally-calculated
virological profile.
Patient reported outcome measures
1] Persisting symptoms and severity (corona symptom checklist)
2] Health-related quality of life (HRQoL, EQ5D)
3] Fatigue (Fatigue Assessment Scale, FAS)
4] Dyspnea (Modified Medical Research Council Dyspnea Scale, mMRC)
5] Cognitive failures in daily life (Cognitive failure questionnaire, CFQ)
6] Return to work (iMTA Productivity Cost Questionnaire, iPCQ)
7] Post-Exertional Malaise (modified sf-DSQ-PEM)
8] Postural orthostatic tachycardia syndrome (Malmo POTS symptom score)
9] Recovery status (Numeric scale and Likert scale)
Background summary
After recovery of the acute phase of COVID-19, a significant proportion of
patients suffer from persistent symptoms, known as long-COVID. This leads to
chronic disability and reduced quality of life. Because the underlying causes
of long-COVID are unclear, there are currently no treatments available. It is
found that several abnormalities, including a dysregulated immune system, an
abnormal anti-viral response and viral persistence, and reactivation of latent
human herpes viruses (HHVs) may contribute to the development of long-COVID.
However, how these abnormalities lead to disease pathogenesis is unknown.
Therefore, in this project we will investigate these abnormalities in more
detail, and link biological data to clinical symptoms, including fatigue,
shortness of breath, muscle aches, headaches, and loss of concentration.
Together, this will bring us closer to understanding the causes of long-COVID.
It will allow us to stratify patients based on these biological components and
will enable us to find possible treatment options for different patient groups.
Study objective
Primary Objective:
Our primary objective is to generate in-depth immune profiles, which include
phenotypic and functional analyses, as well as epigenetic and transcriptomic
analyses, of several B cell, T cell, and monocyte subsets, which relate to
immune activation/dysfunction in patients with long-COVID, and to link these to
clinical parameters. In the lymphoid compartment, B and T cell subsets,
including virus specific T cells, will be analyzed for shifts in subsets, as
well as alterations in activation and/or exhaustion phenotype. B and T cell
receptor signaling will be assessed, as well as cytokine production by
monocytes and lymphocytes. Observed alterations in specific circulating
lymphocyte or monocyte subsets can be further analyzed on transcriptomic and
epigenetic level, using RNA-sequencing (seq) and ATAC-seq, respectively. All
observed alterations will be combined to form unique immune profiles to
discover possible disease-underlying mechanisms, as well as possible targets
for therapy.
Secondary Objective:
Our secondary objective is to generate in-depth virological profiles, to link
aberrations in viral responses or reactivation to clinical parameters. To
explore the possibility that chronic immune activation and dysregulation are
caused or supported by viral persistence, we will measure presence of
SARS-CoV-2 spike protein in serum and specific monocyte populations [22]. Also,
we will determine the profile of SARS-CoV-2-specific B and T cells, and
antibodies present in the circulation of included subjects. We will determine
the breadth of the antibody response by measuring total antibody concentrations
and virus-neutralizing capacity against SARS-CoV-2. To explore whether chronic
immune activation may be caused by a reactivation of latent viruses, we will
determine virus titers of EBV, CMV, and HHV6 in serum/plasma. To link virus
reactivation to the presence of circulating virus-specific antibodies, we will
also determine antibody titers against these latent viruses.
Study design
This study will be a comparative, non-randomized, observational study. Recently
doctor-diagnosed long-COVID patients (<1 year) and long-term ill long-COVID
patients (> 2 years), after PCR or antigen test confirmed initial COVID
infection (WHO long-COVID definition) will be compared with convalescent
controls (CCs).
Study burden and risks
For this study, blood samples will be drawn, of which the risk is minimal.
However, it can cause mild discomfort. This study aims to unravel the
pathophysiology of long-COVID and thus contributes to improve treatment in the
future.
Dr. Molewaterplein 40
Rotterdam 3015 GD
NL
Dr. Molewaterplein 40
Rotterdam 3015 GD
NL
Listed location countries
Age
Inclusion criteria
Long-COVID patients
• Age >= 18 years, <65 years
• Past COVID-19 diagnosis, based on
o Positive PCR
o Positive Sars-Cov2 serology
o Positive rapid antigen test
o Typical clinical syndrome during the first pandemic wave, when testing was
not possible
• Long-COVID-19 diagnosis based on World Health Organisation consensus
diagnosis (*Post COVID-19 condition occurs in individuals with a history of
probable or confirmed SARS CoV-2 infection, usually 3 months from the onset of
COVID-19 with symptoms and that last for at least 2 months and cannot be
explained by an alternative diagnosis. Symptoms may be new onset following
initial recovery from an acute COVID-19 episode or persist from the initial
illness. Symptoms may also fluctuate or relapse over time)*.
o Ref
https://www.who.int/publications/i/item/WHO-2019-nCoV-Post_COVID-19_condition-Cl
inical_case_definition-2021.1
• Overall functioning <70% compared to functioning prior to onset of
Long-COVID/ COVID-19 infection
• Presence of post-exertional malaise
• Provided written informed consent
CC
• Age >= 18 years, <65 years
• Past COVID-19 diagnosis, based on
o Positive PCR
o Positive Sars-Cov2 serology
o Positive rapid antigen test
o Typical clinical syndrome during the first pandemic wave, when testing was
not possible
• No diagnosis of long-COVID, good recovery. Overall functioning >95% compared
to functioning prior COVID-19 infection
• Self-reported general good wellbeing
• Provided written informed consent
Exclusion criteria
Long-COVID patients
• Unable or not willing to provide written informed consent
• Unable to complete written questionnaires in Dutch
• Unable to draw blood for study purposes
• Diagnosis of dementia
• Active treatment with hyperbaric oxygen treatment during study start
• Alternative diagnosis that may explain clinical symptoms
• No re-infection with COVID-19 in the past 3 months
• Suffering from any pre-existing immune-driven disease or use of
anti-inflammatory therapy of any kind (including NSAIDs and steroids) during
the last 3 months
•
CC
• Unable or not willing to provide written informed consent
• Unable to complete written questionnaires in Dutch
• Unable to draw blood for study purposes
• Diagnosis of dementia
• Genetically related to participating patients (e.g. brother/sister/parent)
• Suffering from any immune-driven disease or use of anti-inflammatory therapy
of any kind (including NSAIDs and steroids), including during the last 3 months
• Re-infection with SARS-CoV-2 in the past 3 months.
• Unable or not willing to provide written informed consent
• Unable to complete written questionnaires in Dutch
Design
Recruitment
Followed up by the following (possibly more current) registration
No registrations found.
Other (possibly less up-to-date) registrations in this register
No registrations found.
In other registers
| Register | ID |
|---|---|
| CCMO | NL85275.078.23 |