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Regulation of anti-double-stranded DNA B cells in nonautoimmune mice: localization to the T-B interface of the splenic follicle.

Mandik-Nayak L, Bui A, Noorchashm H, Eaton A, Erikson J - J. Exp. Med. (1997)

Bottom Line: Because the VH3H9 H chain can pair with endogenous L chains to generate anti-single-stranded DNA, anti-dsDNA, and non-DNA B cells, this allowed us to study the regulation of anti-dsDNA B cells in the context of a diverse B cell repertoire.We have identified anti-dsDNA B cells that are located at the T-B interface in the splenic follicle where they have an increased in vivo turnover rate.These anti-dsDNA B cells exhibit a unique surface phenotype suggesting developmental arrest due to antigen exposure.

View Article: PubMed Central - PubMed

Affiliation: The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.

ABSTRACT
Systemic lupus erythematosus (SLE) and the MRL-lpr/lpr murine model for SLE are characterized by the presence of serum anti-double-stranded (ds)DNA antibodies (Abs), whereas nonautoimmune individuals have negligible levels of these Abs. To increase the frequency of anti-DNA B cells and identify the mechanisms involved in their regulation in nonautoimmune mice, we have used Ig transgenes (tgs). In the present study, we used the VH3H9 heavy (H) chain tg which expresses an H chain that was repeatedly isolated from anti-dsDNA Abs from MRL-lpr/lpr mice. Because the VH3H9 H chain can pair with endogenous L chains to generate anti-single-stranded DNA, anti-dsDNA, and non-DNA B cells, this allowed us to study the regulation of anti-dsDNA B cells in the context of a diverse B cell repertoire. We have identified anti-dsDNA B cells that are located at the T-B interface in the splenic follicle where they have an increased in vivo turnover rate. These anti-dsDNA B cells exhibit a unique surface phenotype suggesting developmental arrest due to antigen exposure.

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Increased frequency  of λ+ B cells in VH3H9 tg mice.  Bone marrow (BM; left), spleen  (middle), and lymph node (right)  cells from Tg(−) (open bars) and  VH3H9 (shaded bars) mice were  stained with anti-B220-biotin/ streptavidin-Red670 and anti-λ-FITC or -PE. The percentage of  B220+λ+ cells of total B cells for  spleen and lymph node and percentage of B220+λ+ cells out of total B220+Ig+ cells in the bone marrow  was determined by flow cytometry. The graph shows the mean percentage of B220+λ+ B cells. There is an increased frequency of λ+ B cells in  the VH3H9 spleen (10.1 ± 5.5% versus 5.0 ± 0.8%; P = 0.0011). There  is also a greater frequency of λ+ B cells in the VH3H9 bone marrow  (16.8 ± 7.0% versus 8.3 ± 3.1%; P = 0.0010), but not in the lymph  node (5.4 ± 2.4% versus 4.4 ± 0.7%; P = 0.1139). Interestingly, when  we compare the frequency of λ-expressing B cells within a mouse, the  frequency is highest in the bone marrow and then decreases in the spleen  (VH3H9: 16.8 ± 7.0% versus 10.1 ± 5.5%; P = 0.0087; tg(−): 8.3 ±  3.1% versus 5.0 ± 0.8%; P = 0.0017). The frequency of λs changes only  slightly between the spleen and lymph node in tg(−) mice (5.0 ± 0.8%  versus 4.4 ± 0.7%; P = 0.0251); however, it decreases drastically from  the VH3H9 spleen to the lymph node (10.1 ± 5.5% versus 5.4 ± 2.4%;  P = 0.0034). n = 14 tg(−) mice and n = 13 VH3H9 tg mice.
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Figure 5: Increased frequency of λ+ B cells in VH3H9 tg mice. Bone marrow (BM; left), spleen (middle), and lymph node (right) cells from Tg(−) (open bars) and VH3H9 (shaded bars) mice were stained with anti-B220-biotin/ streptavidin-Red670 and anti-λ-FITC or -PE. The percentage of B220+λ+ cells of total B cells for spleen and lymph node and percentage of B220+λ+ cells out of total B220+Ig+ cells in the bone marrow was determined by flow cytometry. The graph shows the mean percentage of B220+λ+ B cells. There is an increased frequency of λ+ B cells in the VH3H9 spleen (10.1 ± 5.5% versus 5.0 ± 0.8%; P = 0.0011). There is also a greater frequency of λ+ B cells in the VH3H9 bone marrow (16.8 ± 7.0% versus 8.3 ± 3.1%; P = 0.0010), but not in the lymph node (5.4 ± 2.4% versus 4.4 ± 0.7%; P = 0.1139). Interestingly, when we compare the frequency of λ-expressing B cells within a mouse, the frequency is highest in the bone marrow and then decreases in the spleen (VH3H9: 16.8 ± 7.0% versus 10.1 ± 5.5%; P = 0.0087; tg(−): 8.3 ± 3.1% versus 5.0 ± 0.8%; P = 0.0017). The frequency of λs changes only slightly between the spleen and lymph node in tg(−) mice (5.0 ± 0.8% versus 4.4 ± 0.7%; P = 0.0251); however, it decreases drastically from the VH3H9 spleen to the lymph node (10.1 ± 5.5% versus 5.4 ± 2.4%; P = 0.0034). n = 14 tg(−) mice and n = 13 VH3H9 tg mice.

Mentions: λ+ B cells are not only present in the periphery of VH3H9 tg mice, but they are present at a twofold higher frequency than in the tg(−) controls, despite the fact that VH3H9/λ1 B cells are autoreactive and have an increased turnover rate (Fig. 5). What could account for this seemingly surprising result? Pulsing mice with BrdU showed that the increased number of VH3H9/λ B cells is not simply due to their proliferation in the periphery (data not shown). Another possibility is that the VH3H9/λ B cells are positively selected (on some unidentified ligand) in the bone marrow. In support of this, there is a twofold increase in λ+ B cells in the VH3H9 tg bone marrow over tg(−) bone marrow (Fig. 5). Alternatively, the increased λ frequency may be the consequence of receptor editing where λs represent the end result of multiple L chain rearrangement attempts (19, 33). The scenario we favor for an autoreactive Ig in VH3H9 tg mice is that after rearrangement at the κ loci is exhausted, λ rearrangement occurs, completing receptor editing. The λ1+ B cells that are generated are not deleted; rather, they persist in a compromised state.


Regulation of anti-double-stranded DNA B cells in nonautoimmune mice: localization to the T-B interface of the splenic follicle.

Mandik-Nayak L, Bui A, Noorchashm H, Eaton A, Erikson J - J. Exp. Med. (1997)

Increased frequency  of λ+ B cells in VH3H9 tg mice.  Bone marrow (BM; left), spleen  (middle), and lymph node (right)  cells from Tg(−) (open bars) and  VH3H9 (shaded bars) mice were  stained with anti-B220-biotin/ streptavidin-Red670 and anti-λ-FITC or -PE. The percentage of  B220+λ+ cells of total B cells for  spleen and lymph node and percentage of B220+λ+ cells out of total B220+Ig+ cells in the bone marrow  was determined by flow cytometry. The graph shows the mean percentage of B220+λ+ B cells. There is an increased frequency of λ+ B cells in  the VH3H9 spleen (10.1 ± 5.5% versus 5.0 ± 0.8%; P = 0.0011). There  is also a greater frequency of λ+ B cells in the VH3H9 bone marrow  (16.8 ± 7.0% versus 8.3 ± 3.1%; P = 0.0010), but not in the lymph  node (5.4 ± 2.4% versus 4.4 ± 0.7%; P = 0.1139). Interestingly, when  we compare the frequency of λ-expressing B cells within a mouse, the  frequency is highest in the bone marrow and then decreases in the spleen  (VH3H9: 16.8 ± 7.0% versus 10.1 ± 5.5%; P = 0.0087; tg(−): 8.3 ±  3.1% versus 5.0 ± 0.8%; P = 0.0017). The frequency of λs changes only  slightly between the spleen and lymph node in tg(−) mice (5.0 ± 0.8%  versus 4.4 ± 0.7%; P = 0.0251); however, it decreases drastically from  the VH3H9 spleen to the lymph node (10.1 ± 5.5% versus 5.4 ± 2.4%;  P = 0.0034). n = 14 tg(−) mice and n = 13 VH3H9 tg mice.
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Figure 5: Increased frequency of λ+ B cells in VH3H9 tg mice. Bone marrow (BM; left), spleen (middle), and lymph node (right) cells from Tg(−) (open bars) and VH3H9 (shaded bars) mice were stained with anti-B220-biotin/ streptavidin-Red670 and anti-λ-FITC or -PE. The percentage of B220+λ+ cells of total B cells for spleen and lymph node and percentage of B220+λ+ cells out of total B220+Ig+ cells in the bone marrow was determined by flow cytometry. The graph shows the mean percentage of B220+λ+ B cells. There is an increased frequency of λ+ B cells in the VH3H9 spleen (10.1 ± 5.5% versus 5.0 ± 0.8%; P = 0.0011). There is also a greater frequency of λ+ B cells in the VH3H9 bone marrow (16.8 ± 7.0% versus 8.3 ± 3.1%; P = 0.0010), but not in the lymph node (5.4 ± 2.4% versus 4.4 ± 0.7%; P = 0.1139). Interestingly, when we compare the frequency of λ-expressing B cells within a mouse, the frequency is highest in the bone marrow and then decreases in the spleen (VH3H9: 16.8 ± 7.0% versus 10.1 ± 5.5%; P = 0.0087; tg(−): 8.3 ± 3.1% versus 5.0 ± 0.8%; P = 0.0017). The frequency of λs changes only slightly between the spleen and lymph node in tg(−) mice (5.0 ± 0.8% versus 4.4 ± 0.7%; P = 0.0251); however, it decreases drastically from the VH3H9 spleen to the lymph node (10.1 ± 5.5% versus 5.4 ± 2.4%; P = 0.0034). n = 14 tg(−) mice and n = 13 VH3H9 tg mice.
Mentions: λ+ B cells are not only present in the periphery of VH3H9 tg mice, but they are present at a twofold higher frequency than in the tg(−) controls, despite the fact that VH3H9/λ1 B cells are autoreactive and have an increased turnover rate (Fig. 5). What could account for this seemingly surprising result? Pulsing mice with BrdU showed that the increased number of VH3H9/λ B cells is not simply due to their proliferation in the periphery (data not shown). Another possibility is that the VH3H9/λ B cells are positively selected (on some unidentified ligand) in the bone marrow. In support of this, there is a twofold increase in λ+ B cells in the VH3H9 tg bone marrow over tg(−) bone marrow (Fig. 5). Alternatively, the increased λ frequency may be the consequence of receptor editing where λs represent the end result of multiple L chain rearrangement attempts (19, 33). The scenario we favor for an autoreactive Ig in VH3H9 tg mice is that after rearrangement at the κ loci is exhausted, λ rearrangement occurs, completing receptor editing. The λ1+ B cells that are generated are not deleted; rather, they persist in a compromised state.

Bottom Line: Because the VH3H9 H chain can pair with endogenous L chains to generate anti-single-stranded DNA, anti-dsDNA, and non-DNA B cells, this allowed us to study the regulation of anti-dsDNA B cells in the context of a diverse B cell repertoire.We have identified anti-dsDNA B cells that are located at the T-B interface in the splenic follicle where they have an increased in vivo turnover rate.These anti-dsDNA B cells exhibit a unique surface phenotype suggesting developmental arrest due to antigen exposure.

View Article: PubMed Central - PubMed

Affiliation: The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.

ABSTRACT
Systemic lupus erythematosus (SLE) and the MRL-lpr/lpr murine model for SLE are characterized by the presence of serum anti-double-stranded (ds)DNA antibodies (Abs), whereas nonautoimmune individuals have negligible levels of these Abs. To increase the frequency of anti-DNA B cells and identify the mechanisms involved in their regulation in nonautoimmune mice, we have used Ig transgenes (tgs). In the present study, we used the VH3H9 heavy (H) chain tg which expresses an H chain that was repeatedly isolated from anti-dsDNA Abs from MRL-lpr/lpr mice. Because the VH3H9 H chain can pair with endogenous L chains to generate anti-single-stranded DNA, anti-dsDNA, and non-DNA B cells, this allowed us to study the regulation of anti-dsDNA B cells in the context of a diverse B cell repertoire. We have identified anti-dsDNA B cells that are located at the T-B interface in the splenic follicle where they have an increased in vivo turnover rate. These anti-dsDNA B cells exhibit a unique surface phenotype suggesting developmental arrest due to antigen exposure.

Show MeSH
Related in: MedlinePlus