Negative selection of multireactive B cell clones in normal adult mice (original) (raw)
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Normal serum immunoglobulins participate in the selection of peripheral B-cell repertoires
Proceedings of the National Academy of Sciences, 1991
In B-cell development, expression of immunoglobulin heavy-chain variable-region (VH) gene repertoires is determined by genetic mechanisms that favor rearrangement of the most D-proximal genes, resulting in overutilization of the VH7183 gene family early in ontogeny and in differentiating B cells of the adult bone marrow. Maturation of the immune system is accompanied by a decreased expression of VH7183 genes in the peripheral immunocompetent B-cell pool of adult animals. By comparing VH gene family expression in the bone marrow (emergent) and peripheral (available and actual) B-cell repertoires of germ-free and conventionally raised BALB/c mice, we found that peripheral selection of VH gene family utilization does not occur in germ-free animals. Reconstitution of germ-free mice with normal serum immunoglobulins purified from syngeneic donors reestablishes selection of VH7183-expressing B cells. Our results indicate that preimmune B-cell repertoires are selected in normal animals by ...
European Journal of Immunology, 2000
The presence of B cells expressing two B cell receptors (BCR), described in BCRtransgenic, gene-targeted and normal mice, may represent an autoimmune hazard. We generated RAG-2-deficient mice bearing two complete rearranged immunoglobulin transgenes. In these mice most mature resting B cells express chains from the two transgenes. We studied selection of these dual receptor B cells in the presence of self antigens. In spite of the reduced surface density of the anti-self receptor, self-reactive B cells are deleted in the presence of membrane-bound self antigens. In contrast, the presence of soluble self antigen positively selects single receptor B cells expressing the self-reactive receptor. At the periphery these positively selected B cells down-regulate surface IgM expression and become unresponsive. A few dual receptor cells, however, escape tolerance induction. We examined the peripheral fate of the dual receptor B cells and showed that they are poorly selected into the activated B cell compartment and show a poor competitive capacity when in presence of populations of single receptor B cells. These results indicate that peripheral selection contributes to the very low frequencies of dual receptor B cells in normal mice and that multiple safeguard mechanisms operate to minimize the autoimmune hazard that allelically included B cells could represent.
Most peripheral B cells in mice are ligand selected
The Journal of experimental medicine, 1991
Using amplified cDNA and genomic libraries, we have analyzed the VH gene repertoire of pre-B cells and various B cell subsets of conventional mice at the level of VH genes belonging to the J558 VH gene family. The sequence data were evaluated on the basis of a newly established list of 67 J558 VH genes that comprise approximately two-thirds of the J558 VH genes of the murine IgHb haplotype. The results of the analysis demonstrate that VH gene utilization in pre-B cells, although biased to some extent by B cell autonomous VH gene selection, scatters over the whole range of J558 VH genes present in the germline. In contrast, in mature, peripheral B cells comprising long-lived mu + delta high B cells as well as Ly-1 B cells, small overlapping sets of germline VH genes are dominantly expressed. The data indicate that the recruitment of newly generated B cells into the long-lived peripheral B cell pool is mediated through positive selection by internal and/or external antigens. Because o...
B cell repertoire in adult antigen-free and conventional neonatal balb/c mice
European Journal of Immunology, 1989
Early in ontogeny B cells preferentially use VH gene families which are most adjacent to the genes coding for the constant part of the immunoglobulin molecule. In conventional adult mice, however, a random usage of VH gene families has been found. We investigated the role of exogenous antigenic stimulation on this normalization of VH gene usage by B cells. Therefore, we made use of adult germ-free BALB/c mice fed a chemically defined ultrafiltered antigen-free diet (GF-CD) and neonatal conventional BALB/c mice. Both the adult GF-CD and the newborn conventional mice represent situations with minimal exogenous antigenic stimulation. The results obtained with RNA dot blot hybridization with probes specific for the different VH gene families showed in hybridomas from adult GF-CD BALB/c mice a preferential usage of the CH-proximal VH gene family PC7183. In hybridomas from 5-day-old conventional BALB/c mice a less frequent usage of the 5558 VH gene family was found and an increased usage of the PC7183 VH gene family than what would be expected from random usage. Evidence is presented that the RNA giving a positive signal with the PC7183 probe represents functional messages for IgM production.
Differences in the composition of the human antibody repertoire by B cell subsets in the blood
2014
The vast initial diversity of the antibody repertoire is generated centrally by means of a complex series of V(D)J gene rearrangement events, variation in the site of gene segment joining, and TdT catalyzed N-region addition. Although the diversity is great, close inspection has revealed distinct and unique characteristics in the antibody repertoires expressed by different B cell developmental subsets. In order to illustrate our approach to repertoire analysis, we present an in-depth comparison of V(D)J gene usage, hydrophobicity, length, D H reading frame, and amino acid usage between heavy chain repertoires expressed by immature, transitional, mature, memory IgD + , memory IgD − , and plasmacytes isolated from the blood of a single individual. Our results support the view that in both human and mouse, the H chain repertoires expressed by individual, developmental B cell subsets appear to differ in sequence content. Sequencing of unsorted B cells from the blood is thus likely to yield an incomplete or compressed view of what is actually happening in the immune response of the individual. Our findings support the view that studies designed to correlate repertoire expression with diseases of immune function will likely require deep sequencing of B cells sorted by subset.