Antigen presentation kinetics control T cell/dendritic cell interactions and follicular helper T cell generation in vivo
- University of Glasgow, United Kingdom
- University of Zurich, Switzerland
Figures
Figure 1
Immunisation with antigen loaded 200 nm particles induces elevated IgG and can provide protective immunity to influenza virus infection.
Mice were immunised s.c. in the scruff with 100 μg of ovalbumin (OVA) alone or conjugated to 20 nm, 40 nm, 100 nm, 200 nm or 1000 nm particles. 14 days later, serum was tested by ELISA for …
Figure 2
Immunisation with antigen loaded 200 nm particles enhances B cell expansion and germinal centre (GC) B cell responses.
To dissect the cellular responses in detail, MD4 BcR transgenic (Tg) B cells and OT-II TcR Tg T cells and were adoptively transferred into C57BL/6 recipients. Mice were then challenged with OVA-hen …
Figure 3
200 nm particles induce T follicular helper (Tfh) differentiation.
MD4 BcR Tg B cells and OT-II TcR Tg T cells and were adoptively transferred into C57BL/6 recipients. Mice were then challenged with OVA-HEL conjugated to 40 nm or 200 nm particles. Challenges with …
Figure 4
200 nm particulate antigen challenge promotes sustained presentation of peptide/MHCII by dendritic cells (DCs).
C57BL/6 mice were immunized s.c. in the footpad with 100 μg of the model antigen EαGFP alone or covalently conjugated to 40 nm or 200 nm particles. Challenge with PBS was included as a negative …
Figure 5
Peptide/MHCII presentation by B cells.
C57BL/6 mice were immunized s.c. in the footpad with 100 μg of the model antigen EαGFP alone or covalently conjugated to 40 nm or 200 nm particles. Challenge with PBS was included as a negative …
Figure 6
DC and T cell interactions persist after challenge with 200 nm particulate antigen.
DsRed OT-II T cells were adoptively transferred into CD11cYFP recipients and footpad challenged with 100 µg of OVA alone, conjugated to 40 nm or 200 nm particles. Popliteal LNs were imaged at 72 hr. …
Figure 7
Blockade of stage 3 DC/T cell interactions following 200 nm particulate challenge prevents Tfh differentiation.
Disruption of stage 3 DC/T cell interaction was demonstrated by imaging DsRed OT-II T cells adoptively transferred into CD11cYFP recipients. Mice were footpad challenged with 100 µg of OVA …
Figure 8
Cognate interactions between B and T cells are not required for early Tfh differentiation.
OT-II TcR Tg T cells and were adoptively transferred into C57BL/6 or intact MD4 BcR Tg recipients. Mice were then challenged with OVA conjugated to 200 nm particles and 500 µg isotype (mIgG2a) or …
Videos
Video 1
Imaging DC and T cell behaviour after challenge with 200 nm particulate antigen.
DsRed OT-II T cells were adoptively transferred into CD11cYFP recipients and footpad challenged with 100 µg of OVA conjugated to 40 nm or 200 nm particles. Popliteal LNs were imaged at 72 hr. 2 hr …
Video 2
Disruption of DC and T cell interactions using Y3P.
As for Video 1, DsRed OT-II T cells were adoptively transferred into CD11cYFP recipients and footpad challenged with 100 µg of OVA conjugated to 40 nm or 200 nm particles. Popliteal LNs were imaged …
Tables
Table 1
Comparison of particle conjugates
| Particle size | 40 nm | 200 nm |
|---|---|---|
| Antigen dose/mouse | 100 μg | 100 μg |
| Antigen molecules/mouse | 1.3 × 1015 | 1.3 × 1015 |
| Latex dose/mouse | 100 μg | 130 μg |
| Number of particles/mouse | 1.4 × 1012 | 2.9 × 1010 |
| Antigen molecules/particle | 1000 | 20,000 |
| Surface area/particle | 5024 nm2 | 125,600 nm2 |
| Antigen density | 1 per 5 nm2 | 1 per 6 nm2 |
-
Data shows an example of a typical carbodiimide-mediated conjugation of ovalbumin to either 40 or 200 nm. Various parameters relating to a typical s.c. immunisation with 100 μg of protein are shown.
