Evidence for regulation of naturally activated autoreactive B cells

Abstract

A large fraction of naturally activated B cells in the neonate displays degenerate specificity, including reactivity with autoantigens. Transgenic mouse models of autoreactive B cells are mainly concerned with monospecific B cells of high avidity, and the fate of naturally activated autoreactive B cells is still a matter of debate. To pursue this question further, we chose an IgM autoantibody with a recurrent idiotype (Id), i.e. Sp6, because transgenic mice expressing this IgM also were available. In a first approach monoclonal antibodies (mAb) derived from untreated, antigenically stimulated and transgenic mice were used to test whether there were indications for deletion or for Id regulation of naturally activated autoreactive B cells. Over 90% of thymus and spleen cell derived hybridomas from 6-day-old Sp6-transgenic mice were trinitrophenyl (TNP) reactive, carried the Sp6-Id and bound to a panel of self antigens, including mouse albumin. We failed to obtain B cell hybridomas from the thymus of 28-day-old Sp6-transgenic mice. Furthermore, we could not detect any mAb carrying an anti-Sp6 Id, but Sp6 did weakly bind to itself. About 25% of mAb derived from control mice displayed degenerate specificity, the majority of them also were TNP reactive. The Sp6 Id was found at a low frequency and a comparable number of mAb carried an anti-Sp6 Id. Prenatal manipulation at the antigen level (trinitrobenzenesulfonic acid treatment) led to a transient expansion of TNP- and autoreactive mAb. The number of mAb carrying the Sp6 Id was not increased, but mAb carrying an anti-Sp6 Id were observed at high frequency. Those mAb also displayed degenerate specificity. Since Sp6-transgenic mice were perfectly healthy, it is concluded that this particular autoreactive antibody of degenerate specificity cannot be harmful for the developing organism, which may possibly be due to its self-binding capacity. Furthermore, some process of down-regulation was indicated by the absence of B cells expressing the transgene in the thymus of young adult mice. Autoreactivity of untreated and prenatally antigen-treated mice was, in addition, regulated at the Id level. In particular, mAb recognizing the Id of Sp6 were significantly expanded in antigenically stimulated mice. The data were interpreted in the sense that autoreactive B cells appearing early during ontogeny were rather strictly controlled either by (functional) clonal deletion or by idiotypic connectivity.