Unisec

CELL MEDIATED IMMUNITY (CMI) UNISEC TRIAL RESULTS

Access requests to the databases can be made through the Project Manager Ed Schmidt [e.d.l.schmidt@umcg.nl]

All results from these clinical studies will also be made available in scientific publications with the acknowledgement of the EU FP7 contributions to UNISEC.

This database has been made possible through the combined efforts of our UNISEC partners in Work Package 5 and Work Package 6 (BiondVax, Seek, NIPH, NCE, RUG and RKI). We would like to specifically acknowledge the contributions from Stephen Norley (RKI Germany) and Fredrik Oftung (NIPH Norway).

BiondVax Trial Interactive Database 

This Excel file contains a summary of the data obtained from the immunological analyses of samples collected during the course of the BiondVax M-001 vaccine trial. This includes the results of the intracellular cytokine FACS analyses of PBMC after stimulation with peptide pools corresponding to the vaccine (performed by the laboratory of Dr. Stephen Norley, RKI, Berlin) and the results of serological assays (HAI and SRH) performed by the laboratory of Dr. Fredrik Oftung, NIPH, Oslo.

The complete dataset is contained in the sheet named "Main". Each column shows the data associated with a single blood sample. Column headers give the following information: 

  • Subject ID - The unblinded unique subject ID for the participant in the clinical trial
  • Visit No. - The number of the visit to the clinic
  • Date - Date of sampling
  • Treatment -  Recipient of high dose vaccine, low dose vaccine or placebo
  • Rand. Code - Blinded randomisation code used during the analyses
  • Assay ID - Sample ID used during the analyses

Intracellular cytokine FACS analyses.

Cultures were stimulated with the different peptide pools (or controls) before FACS staining. The results are given as the number of cells in the corresponding gate:

  • Events - Total acquired events
  • Lymphocytes - Cells in the lymphocyte gate
  • Viable - Cells in the lymphocyte/viable gate
  • Total CD3+ - Cells in the lymphocyte/viable/CD3+ gate
  • Total CD4+ - Cells in the lymphocyte/viable/CD3+/CD4+ gate
  • Total CD8+ - Cells in the lymphocyte/viable/CD3+/CD8+ gate

For both the CD4+ and CD8+ populations, the number of cells staining positive for the following cytokines/markers are given:

  • IFNg+ - Intracellular interferon gamma
  • IL2+ - Intracellular Interleukin 2
  • TNFa+ - Intracellular tumor necrosis factor alpha
  • CD107a+ - Cell-surface CD107a
  • IL4+ - Intracellular Interleukin 4
  • IL17+ - Intracellular Interleukin 17

Serology

The results (titres) of haemagglutination inhibition (HAI) and single radial haemolysis (SRH) assays using sera from the corresponding blood sampling and four different isolates of influenza are given:  NIBRG14; A/Cal/07/09(H1N1); A/HK/4801/2014(H3N2); B/Brisbane/60/08

Interactive Data Selection

In the worksheet "Selector", one can freely filter the subjects according to visit and treatment. For the ICS FACS analyses, one can filter on the basis of stimulants, markers and cell subsets, and for the serological assays on the basis of assay and virus isolate.

Once the criteria have been selected, pressing the "Extract Selected" button will place a copy of the selected data in the worksheet "Selected" from which it can be copied and analysed further.

The figure shows a snapshot of the interactove BiondVax immunological database containing all the results. It allows different criteria to be selected in different combinations and the selected data to be extracted for further analysis.

 

The figure shows a snapshot of the interactive FACS ICS visualiser for the SEEK data. Each of the little boxes shows the significance of the response to each stimulant (with regard to different cytokine expressions) as a colour block where dark is highly significant and lighter colours less so. The table uses the FDR values calculated by MOMOSA. The subjects are grouped according their patient ID and visit (V2, V4 or V5) and those deemed 'responders' according to the selected criteria (in this case, responses to at least two cytokines to at least two pools in at least one of the post vaccination samples but not in the V2 sample) are highlighted in green. The data to be included in the analysis (cytokine, stimulant) and the criteria for responders can be selected and the table updates automatically. It also calculates the p-value for the Chi2 comparison of the vaccine vs placebo groups automatically.

Seek Trial Interactive Database

This Excel file contains a summary of the data obtained from the immunological analyses of samples collected during the course of the SEEK Flu-V vaccine trial. This includes the results of (i) the intracellular cytokine FACS analyses and (ii) the levels of secreted IFNg using PBMC stimulated with peptide pools corresponding to the vaccine (performed by the laboratory of Dr. Stephen Norley, RKI, Berlin), (iii) the results of assays to measure the serum levels of IgG and IgM and (iv) the results of HAI assays (performed by the laboratory of Dr. Fredrik Oftung, NIPH, Oslo).

The complete dataset is contained in the sheet named "Main". Each column shows the data associated with a single blood sample. Column headers give the following information: 

  • Subject ID - The unblinded unique subject ID for the participant in the clinical trial
  • Visit No. - The number of the visit to the clinic
  • Date - Date of sampling
  • Treatment Group - Treatment group number
  • Treatment - Nature of the vaccine or placebo administered
  • Rand. Code - Blinded randomisation code used during the analyses
  • Assay ID - Sample ID used during the analyses

Intracellular cytokine FACS analyses.

Cultures were stimulated with the different peptide pools (or controls) before FACS staining. The results are given as the number of cells in the corresponding gate:

  • Events - Total acquired events
  • Lymphocytes - Cells in the lymphocyte gate
  • Viable - Cells in the lymphocyte/viable gate
  • Total CD3+ - Cells in the lymphocyte/viable/CD3+ gate
  • Total CD4+ - Cells in the lymphocyte/viable/CD3+/CD4+ gate
  • Total CD8+ - Cells in the lymphocyte/viable/CD3+/CD8+ gate

For both the CD4+ and CD8+ populations, the number of cells staining positive for the following cytokines/markers are given:

  • IFNg+ - Intracellular interferon gamma
  • IL2+ - Intracellular Interleukin 2
  • TNFa+ - Intracellular tumor necrosis factor alpha
  • CD107a+ - Cell-surface CD107a
  • IL4+ - Intracellular Interleukin 4
  • IL17+ - Intracellular Interleukin 17
  •  

IFNg assay

Supernatants from replicate cultures stimulated with the same peptides used for the FACS analysis were assayed for secreted IFNg using a commercial ELISA

Serology

Concentrations of IgG and IgM in the individual serum samples were measured. The results (titres) of haemagglutination inhibition (HAI) assays using four different isolates of influenza are also given:  NIBRG14; A/Cal/07/09(H1N1); A/HK/4801/2014(H3N2); B/Brisbane/60/08

Interactive Data Selection

In the worksheet "Selector", one can freely filter the subjects according to visit and treatment. For the ICS FACS analyses, one can filter on the basis of stimulants, markers (including secreted IFNg) and cell subsets, and for the serological assays on the basis of assay and virus isolate.

Once the criteria have been selected, pressing the "Extract Selected" button will place a copy of the selected data in the worksheet "Selected" from which it can be copied and analysed further.