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. 2017 May 1;214(5):1259–1267. doi: 10.1084/jem.20161533

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© 2017 Kräutler et al.

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Figure 3. — Blocking antigen access, but not Tfh cell help, prevents initiation of PC differentiation in the GC. (A) Schematic of antigen engagement and reception of cognate CD4⁺ Tfh cell help by SW_HEL B cells in unmanipulated GCs (top), plus the mAb-based approaches used to specifically block Tfh cell help (bottom left) or block engagement of intact antigen by the SW_HEL BCR (bottom right). Both HyHEL10* and HyHEL9 bind to HEL^3X, but only HyHEL10* blocks access to the antigen by the SW_HEL BCR. (B) Experimental design for mAb-blocking experiments. SW_HEL.Blimp1^gfp/+ B cells were permitted to form GCs in response to HEL^3X-SRBCs and recipients and then given a single injection of mAbs 2 or 3 d before spleen harvest and analysis on day 9. (C and D) Impact of 3 d of mAb treatment on IgG1⁺ PC-lineage cells in GCs derived from SW_HEL.Blimp1^gfp/+ donor B cells. Representative flow cytometry profiles are shown (C), as well as enumeration of early (Blimp1-GFP^lo IgG1^hi) and late (Blimp1-GFP^hi IgG1^lo) PC-lineage populations in individual recipients (D). (E and F) Impact of 2 d of HyHEL10* treatment analyzed as for C and D. Data from each mAb treatment are representative of two to four independent experiments of five mice per group. Flow cytometry plots are concatenated data from five recipient mice. P-values were calculated using an unpaired Student’s t test. **, 0.001 ≤ P < 0.01.