Surrogate light chain expressing human peripheral B cells produce self-reactive antibodies - PubMed (original) (raw)

Surrogate light chain expressing human peripheral B cells produce self-reactive antibodies

Eric Meffre et al. J Exp Med. 2004.

Abstract

Human B cells that coexpress surrogate and conventional light chains (V-preB+L+) show an unusual heavy and light chain antibody repertoire that display evidence of receptor editing. However, it is unclear whether V-preB+L+ B cells have been silenced by receptor editing or still express autoreactive antibodies. Here we report that 68% of the antibodies expressed by V-preB+L+ B cells are autoreactive. A majority of these autoantibodies are true antinuclear antibodies (ANA), and 50% of the ANAs are also reactive with a diverse group of antigens that include dsDNA, ssDNA, immunoglobulin, insulin, and bacterial lipopolysaccharide. Such antibodies are rarely encountered among conventional B cells. We conclude that V-preB+L+ B cells are a unique subset of normal circulating human B cells that escape central tolerance mechanisms and express self-reactive antibodies including potentially harmful ANAs.

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Figures

Figure 1.

Figure 1.

V-preB+L+ B cell purification scheme. Dot plots show V-preB and Igκ+λ expression on B cells preenriched for V-preB expression using magnetic beads (left), and after the first sort for V-preB+L+ (top right) or conventional V-preB−L+ B cells (bottom right). These populations were subsequently sorted into 96-well plates during a second round of cell sorting.

Figure 2.

Figure 2.

Increased frequency of positively charged amino acids in IgH CDR3s from V-preB+L+ B cells. Pie charts show segment size proportional to the number of clones from V-preB−L+ (left) and V-preB+L+ B cells (right) displaying 0, 1, 2, 3, and 4 or more positively charged amino acids per IgH CDR3. The number of sequences analyzed in each group is indicated in the center.

Figure 3.

Figure 3.

V-preB+L+ B cells express self-reactive antibodies. (A) Antibodies from V-preB+L+ B cells react against HEp-2 cell lysates. ELISAs for anti-HEp-2 cell reactivity using recombinant antibodies from 21 V-preB−L+ (left) and 28 V-preB+L+ B cells (right). The percentage of autoreactive clones for each fraction is indicated. (B) V-preB+L+ antibodies express ANAs. Antibodies from V-preB+L+ B cells show various patterns of ANA including nucleolar (KR9), mitotic spindle apparatus (ED11), speckled (ED20, ED44), and other uncharacterized patterns (ED13, ED38, ED41, ED45), and cytoskeletal reactivity against stress fiber (ED16), vinculin (ED19), and vimentin (ED37). Antibodies isolated from conventional B cells such as EDV-40 do not show ANA staining.

Figure 4.

Figure 4.

V-preB+L+ antibodies are polyreactive. Recombinant antibodies from conventional V-preB−L+ (left) and V-preB+L+ B cells (right) were tested for anti–single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), IgM, insulin, and LPS reactivity by ELISA. Human M55 mAb (dotted line) was included as a positive control polyreactive antibody (22). The percentage of polyreactive clones for each fraction is indicated.

Figure 5.

Figure 5.

Polyreactive antibodies from V-preB+L+ B cells and M55 display similar IgH CDR3 features. Hydrophobic D reading frame and JH6 segments are in olive and brown boxes, respectively. Basic residues are in blue, and acidic amino acids are in red.

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