Primary objective: Dissection of the cellular and serological immune response against SARS-CoV2 with special attention for SARS-CoV2 specific T-cell & B-cell responses and antibody formation against viral proteins as well as for innate…
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Brief title
Condition
- Viral infectious disorders
- Respiratory tract infections
Synonym
Research involving
Sponsors and support
Intervention
Outcome measures
Primary outcome
Absolute counts (relative counts) with comparison to well-defined age-matched
reference values. Based on the first results of the BEAT-COVID-1 *sister
study*, and the ongoing clinical trials of the Innovative Medicines Initiative
(IMI) PERISCOPE program, we expect clear kinetic differences as compared to
age-matched reference values. Comparison between the BEAT-COVID-1 results and
the newly obtained results from this study will potentially explain the reason
of the delays in the antigen-specific immune response in the COVID patients.
Secondary outcome
High throughput sequencing of IG genes (both IGH and IGK-IGL) at the single
cell level of expanded plasma cells and antigen-specific memory B-cells at
consecutive time points in blood. In parallel, the B-cell compartment in
selected nasal scrapes will be evaluated. The transcribed and expressed IGH
isotype usage will be assessed in combination with the full sequence of the
V(D)J exons of both IGH and IGK-IGL genes.
The obtained IG sequences will be mapped against our new *population-matched
IG* (pmIG) data base, which we have developed over the last two years. This is
the first IG database that consists of true genuine germline genes only. This
database is essential to understand how many SHM-induced mutations might have
occurred in the anti-SARS-CoV2 antibodies and to what extend the number and
their position differ within and between the subjects of this study and in
comparison with the COVID patients of the BEAT-COVID-1 project. These
comparative studies will most likely unravel how *fast and easy* antibodies can
be raised in healthy subject compared to COVID patients, where antibody
development seems to be seriously delayed and impaired, suggesting more complex
SHM and selection mechanisms.
In parallel, the TR genes of blood T-lymphocytes and nasal scrapes will be
sequenced to evaluate whether the same or distinct antigen-specific T-cell
responses can be detected in blood and nasal scrapes. This may unravel which
T-cell immune response is most beneficial for eliminating the SARS-CoV2 virus.
Background summary
The ongoing SARS-CoV2 pandemic is rapidly spreading with significant
differences between affected individuals, which are not yet well understood.
Infected individuals show a wide array of clinical symptoms ranging from
asymptomatic or a mild disease course to severe clinical disease requiring
mechanical ventilation in 2 to 3% of cases. Differences in viral dosage and
circumstances of exposure might play a role, but this does not explain the
higher vulnerability of older people and the very low frequency of COVID-19 at
young age. Little is known about the underlying risk factors for severe disease
other than older age, although an association with cardiovascular disease,
diabetes mellitus and obesity has been reported. The ongoing BEAT-COVID-1 study
(ABR no. NL73740.058.20) is focusing on in-depth immune and inflammatory
responses in hospitalized patients, both at COVID departments and at Intensive
Care Units (ICU).
The study presented here focusses on the dissection of the stepwise cellular
and serological immune response in infected individuals with a subclinical or
mild disease course, not requiring hospitalization. The results of this study
will be compared with the results obtained from the BEAT-COVID-1 study.
Understanding differences in immune response against SARS-CoV2 will elucidate
why the disease course can be so heterogeneous.
Study objective
Primary objective: Dissection of the cellular and serological immune response
against SARS-CoV2 with special attention for SARS-CoV2 specific T-cell & B-cell
responses and antibody formation against viral proteins as well as for innate
inflammatory mediators.
- Secondary objective 1: Single cell sequencing of immunoglobulin (IG) genes of
SARS-CoV2-specific plasma cells and memory B-cells with special attention to
anti-Spike antibodies and their capability to block viral binding to human ACE2
proteins.
- Secondary objective 2: Cloning and expression of the selected IG genes for
in-depth studies on antibody reactivity, particularly epitope mapping and
affinity studies.
- Secondary objective 3: Immune profiling of nasal mucosal immune responses.
- Secondary objective 4: Single cell T-cell receptor (TR) gene sequencing of
SARS-CoV2 specific CD4+ and CD8+ T-cells and characterization of dominant
T-cell epitopes.
- Secondary Objective 5: Sequencing of the viral Spike protein, being a major
target of neutralizing antibodies, for mutant analysis, in relation to antibody
responses (1).
- Secondary objective 6: Assessment of activation of the coagulation system in
parallel to the immune response.
- Secondary objective 7: Sequencing of ACE2 and immune-related genes of the
same individuals to understand the potential presence and meaning of
polymorphisms.
Study design
In order to achieve the primary objective and the secondary objectives, a
prospective observational cohort study is proposed in otherwise healthy
individuals (n=10), who have become infected with the SARS-CoV2 virus (= tested
positive by SARS-CoV2 PCR testing of nasal swab, throat swab and/or sputum).
They might have minor or mild clinical symptoms, not requiring hospitalization.
See figure 1 in the study protocol for an extensive description of the study
design.
Study burden and risks
The burden of the study is limited and related to a maximum of 14 donations of
small volumes of blood, nasal swabs, nasal lining fluid (nasosorption filter)
and (optionally) 5 times a nasal inferior turbinate scrape.
The risk of harm is very low. Risks related to venous puncture are slight pain
at puncture site and a hematoma. The amount of blood drawn during the first
week is maximally 90.5 mL, during the first 6 weeks maximally 242 mL, and
during the full period of follow-up (6 months) maximally 294 mL , based on a
maximum duration of SARS-CoV2 positivity of 2 weeks. It is expected that most
subjects will be symptom-free and SARS-CoV2 negative within 2 weeks after
inclusion, implying that generally a total of less than 294 mL will be
collected over a period of 6 months.
Albeit that nasal scrapes have not been studied in patients with airway
infections, no epistaxis was reported in earlier trials with in total
approximately 1,000 healthy donors (incidence < 0.1% based on experience; and
personal communication with S. Jochems). We anticipate that the nasal mucosa
during infection could be more vulnerable and will not perform this in patients
with a history of epistaxis.
Albinusdreef 2
Leiden 2333 ZA
NL
Albinusdreef 2
Leiden 2333 ZA
NL
Listed location countries
Age
Inclusion criteria
• Subject (male or female) tested positive for SARS-CoV2 by PCR (nasal swab,
throat swab and/or sputum);
• Subject is an adult of 18-65 years old;
• Subject is able to understand the PIF document, written in Dutch;
• Subject is able to communicate well with the investigator in the Dutch
language;
• Subject signed informed consent prior to any study-mandated procedure;
• Subject is living in Leiden or direct environment (e.g. within 10 km from
LUMC), allowing for traveling by car, bike or by walking to LUMC (no public
transportation).
Exclusion criteria
• Subject tested negative upon SARS-CoV2 PCR re-analysis of nasal swab at
intake;
• Subject previously fainted before, during or after a medical procedure with
needles;
• Subject received plasma or other blood products during the previous 3 months;
• Subject received a vaccination during the previous 3 months;
• Subject is obese with BMI >=30, based on provide weight and length information;
• Subject has a pre-existing coagulopathy;
• Subject uses medication for a coagulopathy;
• Subject uses medication that suppresses the immune system;
• Subject has an auto-immune disease or another immune disease;
• Subject has another infection in addition to the SARS-CoV2 infection;
• Subject participated in a clinical trial within 6 months prior to this study;
• For women: subject is pregnant or is providing breast-feeding.
Design
Recruitment
Medical products/devices used
metc-ldd@lumc.nl
Followed up by the following (possibly more current) registration
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Other (possibly less up-to-date) registrations in this register
In other registers
| Register | ID |
|---|---|
| CCMO | NL74062.058.20 |