This study will try to objectify if C. Burnetii is able to induce epigenetic changes in monocytes and macrophages, ultimately resulting in a changed cytokine profile. Subsequently, this study will try to link these epigenetic changes to mRNA…
ID
Source
Brief title
Condition
- Ancillary infectious topics
Synonym
Research involving
Sponsors and support
Intervention
Outcome measures
Primary outcome
Primary outcome measures:
•Cytokine concentrations (IL-6, IL-8, IL-10, IFN-γ and TNF-α)
•Epigenetic changes in the different groups of patients (histone modification
H3K4me3).
•Transcriptome analysis
Cytokine concentrations will be measured with ELISA kits while epigenetic
changes will be measured with chromatine immunoprecipitation. Transcriptome
analysis will be conducted through RNA sequencing and biostatic pathway
analysis.
Secondary outcome
Not applicable.
Background summary
Q Fever Fatigue Syndrome (QFS) is a well documented state of prolonged fatigue,
following acute Q fever. Up to 20% of patients that are diagnosed with acute Q
fever will develop QFS, leading to a substantial burden for the affected
patients. This burden constitutes the morbidity, socio-economic weight and
consequences of the disease. Current research focuses mainly on new methods
for diagnosing and treating QFS, while the questions as to why people with QFS
stay fatigued remain unanswered. We would like to investigate the hypothesis
that Coxiella Burnetii (the bacteria that causes Q fever) is able to elicit
epigenetic changes in the monocytes and macrophages that try to clear it. These
epigenetic changes could trigger the cells in such a way that they will secrete
higher levels of pro-inflammatory cytokines, ultimately resulting in a state of
prolonged fatigue (QFS).
Study objective
This study will try to objectify if C. Burnetii is able to induce epigenetic
changes in monocytes and macrophages, ultimately resulting in a changed
cytokine profile. Subsequently, this study will try to link these epigenetic
changes to mRNA expression in the blood of patients and controls.
Study design
An observational case-control study will be performed to determine whether C.
Burnetii is able to induce epigenetic changes in monocytes and macrophages,
ultimately altering their cytokine production. This study will consist of two
main approaches:
•In-vitro experiments will be conducted on Peripheral Blood Mononuclear Cells
(PBMCs) derived from buffy coats of healthy volunteers. PBMCs will be
pre-incubated with C. Burnetii, after which they will be stimulated with
different Pathway Recognition Receptor (PRR) ligands, C. Burnetii or other
bacteria. Once PBMCs are re-stimulated, concentrations of IL-6, IL-8, IL-10 and
TNF-α (cytokines) will be measured in the supernatants and the histone
modification H3K4me3 (related to epigenetic changes) will be investigated
through chromatine immunoprecipitation and qPCR.
•In-vitro experiments will be conducted on blood derived from QFS patients,
Chronic Fatigue Syndrome (CFS) patients, patients with a past C. Burnetii
infection without QFS and healthy controls. PBMC*s will be isolated and
subsequently stimulated with C. burnetii, other bacteria and PRR ligands. After
stimulation, pro-inflammatory cytokine responses (IL-6, TNF- α and IFN-γ) will
be measured. This will give us a preliminary idea of the amount of trained
monocytes in different groups. As soon as this data is analyzed, transcriptome
analysis of leucocytes will be performed in order to link the histone
modifications that might have been found in the former experiment, to mRNA
expression in-vivo.
After informed consent has been obtained, blood will be drawn in 4 EDTA tubes
of 10 ml. RNA of set leucocytes will be stabilized and stored in a freezer
until the RNA can be isolated. RNA sequencing will be done after RNA isolation
and library preparation has been performed according to standard protocols of
the BLUEPRINT-epigenome project and illumine library preparation protocol.
The duration of this study is 2 year. Patients and controls will be recruited
from the Radboudumc and collaborating hospitals.
Study burden and risks
Burden:
•For patients: collection of extra blood, if possible during regular blood
sampling (for CFS and QFS patients), in the form of 4 EDTA tubes of 10 ml
•For controls: the same as for patients, blood samples will only be used for
the research purposes that are described in this study
Risk:
•No risks other than local hematoma are related to venous puncture
Rijnstraat 4
s-Hertogenbosch 5215 EK
NL
Rijnstraat 4
s-Hertogenbosch 5215 EK
NL
Listed location countries
Age
Inclusion criteria
Q Fever Fatigue Syndrome (QFS):
•Diagnosis of QFS according to national LCI-guideline Q fever fatigue syndrome (QFS)
•Score >=40 on the subschale fatigue of the Checklist Individual Strength (CIS)
•Severe functional impairment on Sickness Impact Profile-8 (SIP-8), defined as a SIP total score >=700
•Age >=18;Chronic Fatigue Syndrome (CFS):
•Diagnosed with CFS according to CDC-criteria (www.cdc.gov/cfs)
•Score >=40 on the subschale fatigue of the CIS
•Severe functional impairment on Sickness Impact Profile-8 (SIP-8), defined as a SIP total score >=700
•Age >=18;Cleared acute Q fever without residual symptoms:
•Cleared acute Q fever (without residual symptoms)
•Age >=18;Healthy serologic negative volunteers:
•Negative Q fever serology as tested by immunofluorescence assay (IFA)
•Age >=18
Exclusion criteria
Q Fever Fatigue Syndrome (QFS):
•Use of immunosuppressant drugs during an acute Q fever infection or in the past 3 months
•Pregnancy
•Use of antibiotics that are potentially active against C. Burnetii for at least 4 weeks, after the diagnosis acute Q fever was made;Chronic Fatigue Syndrome (CFS):
•Use of immunosuppressant drugs in the past 3 months
•History of Q fever
•Chronic Q fever patients, according to the Dutch consensus *Chronic Q fever*
•Vaccinated for Q fever
•Pregnancy;Cleared acute Q fever without residual symptoms:
•Use of immunosuppressant drugs during an acute Q fever infection or in the past 3 months
•Chronic Q fever patients, according to the national consensus *Chronic Q fever* [RIVM, Q-koortsvermoeidheidssyndroom]
•Vaccinated for Q fever
•QFS or CFS
•Evident somatic or psychiatric morbidity
•Pregnancy ;Healthy serologic negative volunteers:
•Use of immunosuppressant drugs in the past 3 months
•History of Q fever
•Chronic Q fever patients, according to the national consensus *Chronische Q-koorts*
•Vaccinated for Q fever
•QFS or CFS
•Evident somatic or psychiatric morbidity
•Pregnancy
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 | NL52893.091.15 |