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To investigate the Mycoplasma pneumoniae-specific circulating antibody-secreting cell (ASC) response and Mycoplasma pneumoniae-specific interferon (INF)-γ-secreting T cell response, along with polymerase chain reaction (PCR) and serology, in a cohort of children with community-acquired pneumonia (CAP) and controls.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| CAP cohort |
|
| |
| Healthy control cohort |
|
| |
| Family control cohort | - Family members of index CAP patients. |
|
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Enzyme-linked immunospot (ELISpot) assay [Blood] | Diagnostic Test | The ASC ELISpot will be developed based on the improved methods recently described [Nat Protoc 2013;8:1073-87]. This protocol allows rapid (6-8 h) detection of specific ASCs in small volumes (1-2 ml) of blood. M. pneumoniae protein P1 (50 μl/ml) will be used as antigen. The optimal concentration of coating antigen will be assessed in advance in two-fold serial dilutions for clear spot definition. The M. pneumoniae-specific T cell ELISpot will be developed based on methods recently described [Nat Protoc 2009;4:461-9]. |
| Measure | Description | Time Frame |
|---|---|---|
| Change in numbers of M. pneumoniae-specific ASCs and M. pneumoniae-specific INF-γ-secreting T cells in blood from inclusion (day 0) to 1-month follow-up (day 28) | Enzyme-linked immunospot (ELISpot) assay and flow cytometry | At day 0 (inclusion, disease presentation) and at day 28 (follow-up, disease resolution) |
| Measure | Description | Time Frame |
|---|---|---|
| Change in M. pneumoniae DNA levels in respiratory samples from inclusion (day 0) to 1-month follow-up (day 28) | PCR | At day 0 (inclusion, disease presentation) and at day 28 (follow-up, disease resolution) |
| Change in total and M. pneumoniae-specific antibody levels (immunoglobulin (Ig)G, IgM, IgA) from inclusion (day 0) to 1-month follow-up (day 28) |
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Inclusion Criteria:
CAP cohort:
Healthy control cohort:
- Healthy asymptomatic children of age 3 to 18 years undergoing an elective surgical procedure;
Family control cohort:
- Family members of index CAP patients.
Exclusion Criteria:
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CAP cohort:
There will be a consecutive ongoing recruitment through the project leader or instructed physicians of the department of infectious diseases and emergency in daily clinical practice.
Healthy control cohort:
The project leader will be informed by the local surgeons about a planned elective surgical procedure, unrelated to respiratory tract disease, and will include children during the pre-operation discussion. Samples will be collected by the anaesthesist prior to the start of the surgical procedure, while the child is under general anaesthesia and has an intravenous line. In a subgroup of such children undergoing planned adenotomy, the removed adenoids will be collected after surgery.
Family control cohort:
Family members will be included simultaneously with the index CAP patients.
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| Name | Affiliation | Role |
|---|---|---|
| Patrick M. Meyer Sauteur, MD | University Children's Hospital, Zurich | Principal Investigator |
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Not yet defined.
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|
|
Enzyme-linked immunosorbent assay (ELISA) |
| At day 0 (inclusion, disease presentation) and at day 28 (follow-up, disease resolution) |
| Outcome of community-acquired pneumonia (CAP) assessed by clinical assessment of body temperature (°C) and respiratory rate (per minute) at 1-month follow-up (day 28) | Clinical assessment of body temperature (°C) and respiratory rate (per minute), with worse outcome defined as body temperature more than 38.5°C and respiratory rate according to age more than 40/min for 3 years, more than 34/min for 4-5 years, more than 30/min for 6-12 years, and more than 16/min for 13-18 years. | At day 28 (follow-up) |
| ID | Term |
|---|---|
| D011019 | Pneumonia, Mycoplasma |
| D004194 | Disease |
| ID | Term |
|---|---|
| D009175 | Mycoplasma Infections |
| D009180 | Mycoplasmatales Infections |
| D016905 | Gram-Negative Bacterial Infections |
| D001424 | Bacterial Infections |
| D001423 | Bacterial Infections and Mycoses |
| D007239 | Infections |
| D018410 | Pneumonia, Bacterial |
| D011014 | Pneumonia |
| D012141 | Respiratory Tract Infections |
| D008171 | Lung Diseases |
| D012140 | Respiratory Tract Diseases |
| D010335 | Pathologic Processes |
| D013568 | Pathological Conditions, Signs and Symptoms |
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| ID | Term |
|---|---|
| D058501 | Enzyme-Linked Immunospot Assay |
| D001800 | Blood Specimen Collection |
| ID | Term |
|---|---|
| D003584 | Cytological Techniques |
| D019411 | Clinical Laboratory Techniques |
| D019937 | Diagnostic Techniques and Procedures |
| D003933 | Diagnosis |
| D008919 | Investigative Techniques |
| D004797 | Enzyme-Linked Immunosorbent Assay |
| D007124 | Immunoenzyme Techniques |
| D007118 | Immunoassay |
| D007158 | Immunologic Techniques |
| D007163 | Immunosorbent Techniques |
| D007150 | Immunohistochemistry |
| D015336 | Molecular Probe Techniques |
| D013048 | Specimen Handling |
| D011677 | Punctures |
| D013514 | Surgical Procedures, Operative |
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