Clonal anergy: the universally anergic B lymphocyte.

Abstract

The clonal anergy theory of induction of immunological tolerance states that differentiating B lymphocytes that encounter multivalent antigen at the pre-B to B cell transition stage can receive and store a negative signal, which renders them anergic to later triggering stimuli. The theory was tested by using an anti-IL chain monoclonal antibody, E4, as a model tolerogen.The fluorescence-activated cell sorter was used to select B cellfree cell populations from adult murine bone marrow or newborn spleen, and later, to analyze B cell neogenesis in vitro. The presence of E4 at 21 jg/ml was required to impede the development of normal numbers of B cells with full receptor status. The subsequent capacity of these B cells to respond in vitro to mitogens was assessed in a filler-cell free microculture system that allows single B cells to proliferate and differentiate. Concentrations of E4 far below those required to affect B cell neogenesis had profound inhibitory effects on the subsequent functional capacity of the B cells. In fact, 10-3 jig/ml of E4 markedly impaired both proliferation and antibody formation, and 10' jig/ml, which had no effect on Ig receptor development, abrogated functional ca- It is now well established that immature cells of the B lymphocyte lineage can be rendered immunologically tolerant by certain multivalent antigens both in vitro and in vivo (1-8). Recently, we provided evidence that the stage at which the cell displayed its greatest sensitivity to negative signaling was that of first emergence of the surface membrane immunoglobulin (mIg) receptors, that is, during the pre-B to B cell transition (7). It is possible to introduce tolerogens into the tissues of the developing fetus by transplacental transfer (6) and thus ensure interaction between lymphoid cells and tolerogen at the first appearance of specific antigen binding receptors. When fluoresceinated human gamma globulin (FLU-HGG) was injected into mice at 14.5 days ofgestation, effective B cell tolerance was induced with doses far lower than those required to reduce the number of FLU-specific antigen-binding B cells in the offspring (8). In other words, B lymphocytes expressing a normal range of FLU-binding avidities appeared to emerge from the pre-B cell pool in normal numbers, but were incapable of giving rise to anti-FLU antibody-forming cell (AFC) clones when challenged with a B cell mitogen or a T lymphocyte-independent antigen. This suggested that interaction between the newly emerged anti-FLU receptors and antigen had neither impeded the subsequent development of a standard complement of mIg nor directly led to the death of the anti-FLU B cell. Rather, the cell, though still alive and capable ofbinding antigen, appeared to have received some signal rendering it incapable of responding to appropriate triggering stimuli. We termed this phenomenon clonal anergy.This conclusion was dependent on a technology that enumerated and characterized FLU-binding B cells, by counting cells adhering specifically to thin layers of hapten...

Balancing immunity and tolerance: deleting and tuning lymphocyte repertoires.

“…Self-tolerance can also be achieved by functionally altering self-reactive cells instead of physically eliminating them. In B cells, a period of exposure to a relatively weak, costimulator-deficient antigenic stimulus, such as soluble anti-immunoglobulin antibody (98), serum lysozyme (99, 100), or singlestranded DNA (37,101), renders the B cell much more difficult to activate into proliferation and antibody secretion by a subsequent immunogenic antigen challenge. Anergy in B cells reflects retuning of the cells' antigen receptors.…”

“…In the B-cell lineage, immature B cells do not express the costimulatory CD21 receptor for C3d (16,196), and antigen stimulation fails to trigger CD86/B7.2 (J. G. Cyster and C.C.G., unpublished observations). Moreover, immature B cells that have bound antigen lose responsiveness to LPS (16,98,197,198). In mature B cells, sudden antigen stimulation provokes display of CD86/B7.2, but after two days CD86/B7.2 expression returns to baseline despite sustained antigenic stimulation, and it is not elevated on B cells that have been chronically stimulated by antigen (28,61,192,193 (199).…”

Novel Roles for Lyn in B Cell Migration and Lipopolysaccharide Responsiveness Revealed Using Anti-Double-Stranded DNA Ig Transgenic Mice

“…Given that signals through Ig are reported to impair proliferation to LPS (35), as does the absence of Lyn, lyn Ϫ/Ϫ anti-dsDNA B cells might be expected to exhibit profound deficits in their response to LPS. However, remarkably, Lyn deficiency actually restores their ability to proliferate in the presence of LPS (Fig.…”

“…For example, B cells that have received an initial signal through the BCR appear hyporesponsive to B cell mitogens such as LPS (35). To see whether Lyn is involved in this process, we tested the response of lyn Ϫ/Ϫ anti-dsDNA B cells to LPS stimulation.…”

B Cells from Mice Prematurely Expressing Human Complement Receptor Type 2 Are Unresponsive to T-Dependent Antigens

“…Classical B cell anergy has been shown to produce B cells that are nonresponsive to LPS stimulation in culture (15,17,39,40). To address whether the B cells that express hCR2 resemble anergic cells in this respect, splenic B cells isolated from hCR2 high or hCR2 Ϫ littermates were incubated with LPS in vitro for 72 h. B cell numbers were measured, and expression of B cell activation markers was assessed.…”