Experiment Overview
| Repository ID: | FR-FCM-Z24V | Experiment name: | MiniHA comparison; an assessment of B cell responses post immunization | MIFlowCyt score: | 100.00% |
| Primary researcher: | Joost Vaneman | PI/manager: | Joost Vaneman | Uploaded by: | Joost Vaneman |
| Experiment dates: | 2016-01-14 - 2016-01-14 | Dataset uploaded: | Jun 2019 | Last updated: | Jul 2019 |
| Keywords: | [B cells] [Influenza] [Germinal center] [vaccine candidate screening] | Manuscripts: | |||
| Organizations: |
Janssen Vaccines & Prevention B.V., Viral Vaccines, Leiden, (The Netherlands)
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| Purpose: | The main purpose of this experiment was to determine whether the magnitude of the GC B-cell frequencies present in the iliac lymph nodes of mice, measured 4 days post boost, reflects the protective efficacy associated with the administered universal flu vaccine candidate antigens. | ||||
| Conclusion: | Animals immunized with UFV#4157 or UFV#4650 expressed significantly higher frequencies of HA-specific Germinal center B cells (CD4- CD19+ CD95+ GL7+ 5070+) and HA specific non germinal center B cells (CD4- CD19+ CD95- GL7- 5070+) in comparison to the protective UFV#4900 antigen. These results support the hypothesis that poorly protecting HA-derived antigens recall more germinal center responses then protective candidates. | ||||
| Comments: | We compared (antigen specific) GC responses (CD4- CD19+ CD95+ GL7+ ) induced by structurally closely related antigens with known differences in the level of protective efficacy. For this, we immunized cohorts of mice using the same prime-boost scheme with three mini-HA antigens: monomeric UFV#4157, mostly dimeric UFV#4650 and trimeric UFV#4900 at 30 µg/dose. These mini-HA's are all derived from the same H1 A/Brisbane/59/07 HA stem and do not contain the immunodominant head of the HA molecule. In heterologous and heterosubtypic influenza challenges each candidate confers different levels of protection, ranging from minimal to partial to broad protection against heterologous group 1 strains for UFV#4157, UFV#4650 and UFV#4900 respectively. | ||||
| Funding: | N/A | ||||
| Quality control: | Before the experimental samples and compensation controls were acquired on the LSR Fortessa, a CST check was performed to verify the cytometers performance. CST Check Passed. | ||||
Experiment variables
| Timepoints | |
|---|---|
| · T=25 days (4 days post boost) | 2316 HD in 5070 detection_A5_A05_046.fcs · 2316 HD in 5070 detection_B5_B05_047.fcs · 2316 HD in 5070 detection_C5_C05_048.fcs · 2316 HD in 5070 detection_D5_D05_049.fcs · 2316 HD in 5070 detection_E5_E05_050.fcs · 2316 HD in 5070 detection_F5_F05_051.fcs · 2316 HD in 5070 detection_G5_G05_052.fcs · 2316 HD in 5070 detection_H5_H05_053.fcs · 4157 in 5070 detection_A2_A02_022.fcs · 4157 in 5070 detection_B2_B02_023.fcs · 4157 in 5070 detection_C2_C02_024.fcs · 4157 in 5070 detection_D2_D02_025.fcs · 4157 in 5070 detection_E2_E02_026.fcs · 4157 in 5070 detection_F2_F02_027.fcs · 4157 in 5070 detection_G2_G02_028.fcs · 4157 in 5070 detection_H2_H02_029.fcs · 4650 in 5070 detection_A3_A03_030.fcs · 4650 in 5070 detection_B3_B03_031.fcs · 4650 in 5070 detection_C3_C03_032.fcs · 4650 in 5070 detection_D3_D03_033.fcs · 4650 in 5070 detection_E3_E03_034.fcs · 4650 in 5070 detection_F3_F03_035.fcs · 4650 in 5070 detection_G3_G03_036.fcs · 4650 in 5070 detection_H3_H03_037.fcs · 4900 in 5070 detection_A4_A04_038.fcs · 4900 in 5070 detection_B4_B04_039.fcs · 4900 in 5070 detection_C4_C04_040.fcs · 4900 in 5070 detection_D4_D04_041.fcs · 4900 in 5070 detection_E4_E04_042.fcs · 4900 in 5070 detection_F4_F04_043.fcs · 4900 in 5070 detection_G4_G04_044.fcs · 4900 in 5070 detection_H4_H04_045.fcs · PBS in 5070 detection_A1_A01_014.fcs · PBS in 5070 detection_B1_B01_015.fcs · PBS in 5070 detection_C1_C01_016.fcs · PBS in 5070 detection_D1_D01_017.fcs · PBS in 5070 detection_E1_E01_018.fcs · PBS in 5070 detection_F1_F01_019.fcs · PBS in 5070 detection_G1_G01_020.fcs · PBS in 5070 detection_H1_H01_021.fcs |
| Doses | |
|---|---|
| · PBS (Mock Control) | PBS in 5070 detection_A1_A01_014.fcs · PBS in 5070 detection_B1_B01_015.fcs · PBS in 5070 detection_C1_C01_016.fcs · PBS in 5070 detection_D1_D01_017.fcs · PBS in 5070 detection_E1_E01_018.fcs · PBS in 5070 detection_F1_F01_019.fcs · PBS in 5070 detection_G1_G01_020.fcs · PBS in 5070 detection_H1_H01_021.fcs |
| · 60µg FL H1#2316 | 2316 HD in 5070 detection_A5_A05_046.fcs · 2316 HD in 5070 detection_B5_B05_047.fcs · 2316 HD in 5070 detection_C5_C05_048.fcs · 2316 HD in 5070 detection_D5_D05_049.fcs · 2316 HD in 5070 detection_E5_E05_050.fcs · 2316 HD in 5070 detection_F5_F05_051.fcs · 2316 HD in 5070 detection_G5_G05_052.fcs · 2316 HD in 5070 detection_H5_H05_053.fcs |
| · 30µg UFV#4157 | 4157 in 5070 detection_A2_A02_022.fcs · 4157 in 5070 detection_B2_B02_023.fcs · 4157 in 5070 detection_C2_C02_024.fcs · 4157 in 5070 detection_D2_D02_025.fcs · 4157 in 5070 detection_E2_E02_026.fcs · 4157 in 5070 detection_F2_F02_027.fcs · 4157 in 5070 detection_G2_G02_028.fcs · 4157 in 5070 detection_H2_H02_029.fcs |
| · 30µg UFV#4650 | 4650 in 5070 detection_A3_A03_030.fcs · 4650 in 5070 detection_B3_B03_031.fcs · 4650 in 5070 detection_C3_C03_032.fcs · 4650 in 5070 detection_D3_D03_033.fcs · 4650 in 5070 detection_E3_E03_034.fcs · 4650 in 5070 detection_F3_F03_035.fcs · 4650 in 5070 detection_G3_G03_036.fcs · 4650 in 5070 detection_H3_H03_037.fcs |
| · 30µg UFV#4900 | 4900 in 5070 detection_A4_A04_038.fcs · 4900 in 5070 detection_B4_B04_039.fcs · 4900 in 5070 detection_C4_C04_040.fcs · 4900 in 5070 detection_D4_D04_041.fcs · 4900 in 5070 detection_E4_E04_042.fcs · 4900 in 5070 detection_F4_F04_043.fcs · 4900 in 5070 detection_G4_G04_044.fcs · 4900 in 5070 detection_H4_H04_045.fcs |
| Sample Type | |
|---|---|
| · iliac lymph nodes | 2316 HD in 5070 detection_A5_A05_046.fcs · 2316 HD in 5070 detection_B5_B05_047.fcs · 2316 HD in 5070 detection_C5_C05_048.fcs · 2316 HD in 5070 detection_D5_D05_049.fcs · 2316 HD in 5070 detection_E5_E05_050.fcs · 2316 HD in 5070 detection_F5_F05_051.fcs · 2316 HD in 5070 detection_G5_G05_052.fcs · 2316 HD in 5070 detection_H5_H05_053.fcs · 4157 in 5070 detection_A2_A02_022.fcs · 4157 in 5070 detection_B2_B02_023.fcs · 4157 in 5070 detection_C2_C02_024.fcs · 4157 in 5070 detection_D2_D02_025.fcs · 4157 in 5070 detection_E2_E02_026.fcs · 4157 in 5070 detection_F2_F02_027.fcs · 4157 in 5070 detection_G2_G02_028.fcs · 4157 in 5070 detection_H2_H02_029.fcs · 4650 in 5070 detection_A3_A03_030.fcs · 4650 in 5070 detection_B3_B03_031.fcs · 4650 in 5070 detection_C3_C03_032.fcs · 4650 in 5070 detection_D3_D03_033.fcs · 4650 in 5070 detection_E3_E03_034.fcs · 4650 in 5070 detection_F3_F03_035.fcs · 4650 in 5070 detection_G3_G03_036.fcs · 4650 in 5070 detection_H3_H03_037.fcs · 4900 in 5070 detection_A4_A04_038.fcs · 4900 in 5070 detection_B4_B04_039.fcs · 4900 in 5070 detection_C4_C04_040.fcs · 4900 in 5070 detection_D4_D04_041.fcs · 4900 in 5070 detection_E4_E04_042.fcs · 4900 in 5070 detection_F4_F04_043.fcs · 4900 in 5070 detection_G4_G04_044.fcs · 4900 in 5070 detection_H4_H04_045.fcs · PBS in 5070 detection_A1_A01_014.fcs · PBS in 5070 detection_B1_B01_015.fcs · PBS in 5070 detection_C1_C01_016.fcs · PBS in 5070 detection_D1_D01_017.fcs · PBS in 5070 detection_E1_E01_018.fcs · PBS in 5070 detection_F1_F01_019.fcs · PBS in 5070 detection_G1_G01_020.fcs · PBS in 5070 detection_H1_H01_021.fcs |
