Autoreactive B cells in the marginal zone that express dual receptors - PubMed (original) (raw)

Autoreactive B cells in the marginal zone that express dual receptors

Yijin Li et al. J Exp Med. 2002.

Abstract

Allotype and isotype exclusion is a property of most lymphocytes. The reason for this property is not known but it guarantees a high concentration of a single receptor, and threshold numbers of receptors may be required for efficient positive and negative selection. Receptor editing compromises exclusion by sustaining recombination even after a functional receptor is formed. Consequently, B cells expressing multiple receptors arise. We have studied such B cells in which one of the two receptors is anti-self, and find that these partially autoreactive B cells accumulate in the marginal zone. The restriction of these cells in this location may help to prevent them from undergoing diversification and developing into fully autoreactive B cells.

PubMed Disclaimer

Figures

Figure 1.

Figure 1.

FACS® analysis of mature B cells in anti-DNA H chain sd-tg mice. (A) Splenic and bone marrow cells were isolated from 3H9H, 3H9H/56R, and 3H9H/56R/76R sd-tg mice. Cells were stained with antibodies against B220, CD43, IgM, and IgD. All analyses were performed on the lymphocyte-gated population. IgM+IgD+ cells were plotted from the B220+CD43−-gated population. (B) Recognition of transgenic H chain–bearing cells by the anti-idiotypic Ab, 1.209, which recognizes the 3H9 H chain in combination with most L-chain. Cells were double-stained with anti-B220 and 1.209. Data were plotted and percentages calculated from the lymphocyte-gated population. These results are representative of four independent experiments.

Figure 2.

Figure 2.

κ and λ expression in spleen cells of mice with H chain of different DNA binding affinities. (A) Spleen cells from BALB/c, 3H9, 3H9H/56R, 3H9H/56R/76R sd-tg mice were stained with anti-κ and -λ. Cells were gated on a lymphoid gate, and percentages of κ+, λ+, and κ/λ+ are indicated. (B) 3H9H/56R mice have a large population of κ/λ double-positive B cells. Spleen cells from 3H9H/56R and the nontransgenic littermate were stained with anti-B220, κ, and λ. Dead cells were excluded by propidium iodide staining. Percentages of κ+, λ+, and κ/λ+ cells in a B220+ gate are indicated. Representative of experiments using three different mice of each kind. (C) Surface IgM expression of κ/λ double-positive B cells are higher than most of the κ single-positive B cells in 3H9H/56R mice. Histograms of IgM expression are shown for the κ/λ double-positive (R4 and thin line) and κ single-positive (R5 and bold line) population.

Figure 2.

Figure 2.

κ and λ expression in spleen cells of mice with H chain of different DNA binding affinities. (A) Spleen cells from BALB/c, 3H9, 3H9H/56R, 3H9H/56R/76R sd-tg mice were stained with anti-κ and -λ. Cells were gated on a lymphoid gate, and percentages of κ+, λ+, and κ/λ+ are indicated. (B) 3H9H/56R mice have a large population of κ/λ double-positive B cells. Spleen cells from 3H9H/56R and the nontransgenic littermate were stained with anti-B220, κ, and λ. Dead cells were excluded by propidium iodide staining. Percentages of κ+, λ+, and κ/λ+ cells in a B220+ gate are indicated. Representative of experiments using three different mice of each kind. (C) Surface IgM expression of κ/λ double-positive B cells are higher than most of the κ single-positive B cells in 3H9H/56R mice. Histograms of IgM expression are shown for the κ/λ double-positive (R4 and thin line) and κ single-positive (R5 and bold line) population.

Figure 2.

Figure 2.

κ and λ expression in spleen cells of mice with H chain of different DNA binding affinities. (A) Spleen cells from BALB/c, 3H9, 3H9H/56R, 3H9H/56R/76R sd-tg mice were stained with anti-κ and -λ. Cells were gated on a lymphoid gate, and percentages of κ+, λ+, and κ/λ+ are indicated. (B) 3H9H/56R mice have a large population of κ/λ double-positive B cells. Spleen cells from 3H9H/56R and the nontransgenic littermate were stained with anti-B220, κ, and λ. Dead cells were excluded by propidium iodide staining. Percentages of κ+, λ+, and κ/λ+ cells in a B220+ gate are indicated. Representative of experiments using three different mice of each kind. (C) Surface IgM expression of κ/λ double-positive B cells are higher than most of the κ single-positive B cells in 3H9H/56R mice. Histograms of IgM expression are shown for the κ/λ double-positive (R4 and thin line) and κ single-positive (R5 and bold line) population.

Figure 3.

Figure 3.

Increase of MZ B cells in the H chain transgenic mice. (A) Spleen cells from BALB/c, 3H9, 3H9H/56R, 3H9H/56R/76R sd-tg mice were stained with anti-B220, -CD21, and -CD23. Percentages of CD21high and CD23low populations in the B220+ gate are indicated. (B) Increase of MZ B cells in the H chain sd-tg mice shown by immunohistochemistry. Spleen sections of each mouse were stained with anti-IgM that stains both MZ and follicular B cell (blue) and MOMA-1 that detects metallophilic marginal macrophage (red).

Figure 4.

Figure 4.

The κ/λ double-positive B cells in 3H9H/56R are MZ B cells. Histograms of CD23, CD21, and IgD expression are shown for the κ/λ double-positive (R4 and thin line) and κ single-positive (R5 and bold line) population. Representative of experiments using three different mice.

Figure 5.

Figure 5.

Allelic exclusion by the 3H9H/56R H chain transgene. Spleen cells from 3H9H/56R mice and the nontransgenic littermate were stained with anti-CD19, -IgMa, and -IgMb. Percentages of IgMa+ (the transgenic allele) and IgMb+ (the endogenous allele) cells in the CD19+ gate are indicated. Representative of two independent experiments using a different mouse of each kind.

Figure 6.

Figure 6.

Detection of Id-positive B cells in 3H9H/56R mice. (A) The majority of Id-positive B cells in 3H9H/56R mice are in the MZ. Spleen cells from 3H9H/56R mice and the nontransgenic littermate were stained with anti-B220, -CD21, and 1.209 anti-Id antibodies. Percentages of CD21high and Id-positive cells in the B220+ gate are indicated. Representative of experiments using three different mice of each kind. (B) The κ/λ double-positive B cells in 3H9H/56R are Id positive. Histogram of 1.209 anti-Id staining are shown for the κ/λ double-positive (R4 and thin line) and κ single-positive (R5 and bold line) populations. Representative of experiments using three different mice.

Figure 6.

Figure 6.

Detection of Id-positive B cells in 3H9H/56R mice. (A) The majority of Id-positive B cells in 3H9H/56R mice are in the MZ. Spleen cells from 3H9H/56R mice and the nontransgenic littermate were stained with anti-B220, -CD21, and 1.209 anti-Id antibodies. Percentages of CD21high and Id-positive cells in the B220+ gate are indicated. Representative of experiments using three different mice of each kind. (B) The κ/λ double-positive B cells in 3H9H/56R are Id positive. Histogram of 1.209 anti-Id staining are shown for the κ/λ double-positive (R4 and thin line) and κ single-positive (R5 and bold line) populations. Representative of experiments using three different mice.

Figure 7.

Figure 7.

Loss of immature and mature/recirculating B cells and absence of κ/λ and Id-positive B cells in the bone marrow of 3H9H/56R mice. (A and B) Bone marrow cells from 3H9H/56R mice and the nontransgenic littermate were stained with anti-B220, -IgM, and -IgD. Percentages of B cells in the subsets of IgMlowB220low, IgMhighB220low, IgM+B220high, IgDlowB220low, and IgDhighB220high are indicated. (C) Bone marrow cells gated from IgDlowB220low and IgDhighB220high are shown for their light chain expression. Percentages of κ+, λ+, and κ/λ+ cells are indicated. (D) Bone marrow cells are stained with anti-B220, -IgM, and 1.209. Percentages of Id+, IgM+, and Id/IgM+ cells in a B220+ gate are indicated.

Figure 8.

Figure 8.

Increase of κ/λ double-positive, MZ, and Id-positive B cells in the spleen of 3H9H/56R mice with age. Spleen cells from 3H9H/56R mice of age 5, 7.5, and 11 wk were stained with anti-B220, -κ, -λ, -CD21, -CD23, and 1.209 anti-Id antibodies. Percentages of κ/λ double-positive B cell, CD21high and CD23low MZ B cells, and Id-positive cells in the B220+ gate are indicated.

Similar articles

Cited by

References

    1. Alt, F.W., V. Enea, A.L. Bothwell, and D. Baltimore. 1980. Activity of multiple light chain genes in murine myeloma cells producing a single, functional light chain. Cell. 21:1–12. - PubMed
    1. Coleclough, C., R.P. Perry, K. Karjalainen, and M. Weigert. 1981. Aberrant rearrangements contribute significantly to the allelic exclusion of immunoglobulin gene expression. Nature. 290:372–378. - PubMed
    1. Ramsden, D.A., and G.E. Wu. 1991. Mouse kappa light-chain recombination signal sequences mediate recombination more frequently than do those of lambda light chain. Proc. Natl. Acad. Sci. USA. 88:10721–10725. - PMC - PubMed
    1. Yamagami, T., E. ten Boekel, C. Schaniel, J. Andersson, A. Rolink, and F. Melchers. 1999. Four of five RAG-expressing JCkappa−/− small pre-BII cells have no L chain gene rearrangements: detection by high-efficiency single cell PCR. Immunity. 11:309–316. - PubMed
    1. Retter, M.W., and D. Nemazee. 1998. Receptor editing occurs frequently during normal B cell development. J. Exp. Med. 188:1231–1238. - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources