To examine how B cell receptor affinity affects clonal selection in thymus-independent type 2 (TI-2) immune responses, we produced mice with antibodies that showed a 40-fold difference in affinity for the hapten (4-hydroxy-3-nitrophenyl)acetyl (NP). The difference in the responses of high- and low-affinity B cells to NP-Ficoll was only twofold. However, in competition experiments only the high-affinity B cells responded to antigen. CD19 deficiency increased the affinity threshold of TI-2 responses, whereas Lyn deficiency enhanced clonal expansion but abrogated B cell terminal differentiation. Thus, in TI-2 immune responses, large differences in affinity produce only small differences in the intrinsic ability of B cells to respond to antigen, and selection for high-affinity clones is due to clonal competition during the earliest stages of the response.
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We thank E. Besmer and members of the Nussenzweig lab for helpful comments on the manuscript, Nai-Ying Zheng for histology and Frank Isdell and Michelle Genova for FACS. We also thank C. Lowell, M. Neuberger, R. Pelanda, R. Rickert and T. Tedder for constructs, reagents and mice. Supported by NIH MSTP grant GM07739 (T. Y. S.), HHMI and grants from the Leukemia Society and NIH to M.C.N.
The authors declare no competing financial interests.
Flow cytometric analysis of immunized (d5) (a) CD22-/- B1-8hi and (b) CD22-/- B1-8lo mice and unimmunized controls (d0). The first row shows B220/Igλ staining with λ + B cells indicated within the gate. Plots in the second row are B220+-gated and show GC cells within the gate (Fas+GL7+ cells). Plasmacyte differentiation is shown in the third row by B220/Syndecan-1 staining. All plots are lymphocyte-gated with additional gating as specified; numbers indicate percentages of cells within the designated gate. (GIF 42 kb)
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Shih, T., Roederer, M. & Nussenzweig, M. Role of antigen receptor affinity in T cell–independent antibody responses in vivo. Nat Immunol 3, 399–406 (2002). https://doi.org/10.1038/ni776
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