Positive selection of B10 cells is determined by BCR specificity and signaling strength.

B10 cells, a regulatory B cell subset, negatively regulate immune responses in an IL-10-dependent manner. However, the mechanism of B10 cell development is unclear. We found that B10 cells mainly identified self-antigens. TgVH3B4 transgenic mice, whose VH was derived from an actin-reactive natural antibody, exhibit elevated numbers of actin-binding B10 cells. Immunization of TgVH3B4 mice with actin induced elevated B10 cell numbers in an antigen-specific manner, indicating positive selection of B10 cells by self-antigens. Furthermore, higher BCR signaling strength facilitated B10 cell development. We also observed that actin-reactive IgG levels were unchanged in TgVH3B4 mice after immunization with actin in contrast to the elevated OVA-reactive IgG level after immunization with OVA, indicating that B10 cells acted in an antigen-specific manner to inhibit the immune response. Our data demonstrate for the first time that B10 cells are positively selected by self-reactivity and that higher BCR signaling strength promotes B10 cell development.