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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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VH3H9/λ B cells accumulate at the T–B  interface. Spleen sections from Tg(−) (left) and  VH3H9 (right) mice were stained with anti-λ-AP and  either anti-B220-biotin (top) or anti-CD4-biotin (bottom). A similar staining pattern was observed when  anti-CD8-biotin was included with anti-CD4-biotin  (data not shown). Avidin-HRP was used as a secondary step for the biotinylated reagents. HRP and AP  were developed using the substrates, 3-amino-9-ethyl-carbazole (red) and Fast-Blue BB base (blue), respectively. These are representative sections from n = 15  Tg(−) and n = 19 VH3H9 mice. Original magnification: 100.
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Figure 4: VH3H9/λ B cells accumulate at the T–B interface. Spleen sections from Tg(−) (left) and VH3H9 (right) mice were stained with anti-λ-AP and either anti-B220-biotin (top) or anti-CD4-biotin (bottom). A similar staining pattern was observed when anti-CD8-biotin was included with anti-CD4-biotin (data not shown). Avidin-HRP was used as a secondary step for the biotinylated reagents. HRP and AP were developed using the substrates, 3-amino-9-ethyl-carbazole (red) and Fast-Blue BB base (blue), respectively. These are representative sections from n = 15 Tg(−) and n = 19 VH3H9 mice. Original magnification: 100.

Mentions: Lymphocytes are organized into discrete structures within the splenic architecture. Resting T and B cells segregate into the inner periarteriolar lymphoid sheath (inner PALS or T cell zone) and outer PALS (B cell follicle) within the spleen. To address where the short-lived anti-dsDNA B cells are located within the splenic architecture, spleen sections from tg(−) and VH3H9 tg mice were stained with anti-λ, anti-B220, and anti-CD4 to mark B and T zones, respectively (Fig. 4). In tg(−) mice, λ+ B cells are scattered throughout the B cell area in the follicle as well as in the red pulp, but are not detected within the T zone. In contrast, the λ+ anti-dsDNA B cells in the VH3H9 tg mouse are found clustered at the T–B interface in the B cell follicle, as well as in the T cell zone (Fig. 4).


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)

VH3H9/λ B cells accumulate at the T–B  interface. Spleen sections from Tg(−) (left) and  VH3H9 (right) mice were stained with anti-λ-AP and  either anti-B220-biotin (top) or anti-CD4-biotin (bottom). A similar staining pattern was observed when  anti-CD8-biotin was included with anti-CD4-biotin  (data not shown). Avidin-HRP was used as a secondary step for the biotinylated reagents. HRP and AP  were developed using the substrates, 3-amino-9-ethyl-carbazole (red) and Fast-Blue BB base (blue), respectively. These are representative sections from n = 15  Tg(−) and n = 19 VH3H9 mice. Original magnification: 100.
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Related In: Results  -  Collection

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Figure 4: VH3H9/λ B cells accumulate at the T–B interface. Spleen sections from Tg(−) (left) and VH3H9 (right) mice were stained with anti-λ-AP and either anti-B220-biotin (top) or anti-CD4-biotin (bottom). A similar staining pattern was observed when anti-CD8-biotin was included with anti-CD4-biotin (data not shown). Avidin-HRP was used as a secondary step for the biotinylated reagents. HRP and AP were developed using the substrates, 3-amino-9-ethyl-carbazole (red) and Fast-Blue BB base (blue), respectively. These are representative sections from n = 15 Tg(−) and n = 19 VH3H9 mice. Original magnification: 100.
Mentions: Lymphocytes are organized into discrete structures within the splenic architecture. Resting T and B cells segregate into the inner periarteriolar lymphoid sheath (inner PALS or T cell zone) and outer PALS (B cell follicle) within the spleen. To address where the short-lived anti-dsDNA B cells are located within the splenic architecture, spleen sections from tg(−) and VH3H9 tg mice were stained with anti-λ, anti-B220, and anti-CD4 to mark B and T zones, respectively (Fig. 4). In tg(−) mice, λ+ B cells are scattered throughout the B cell area in the follicle as well as in the red pulp, but are not detected within the T zone. In contrast, the λ+ anti-dsDNA B cells in the VH3H9 tg mouse are found clustered at the T–B interface in the B cell follicle, as well as in the T cell zone (Fig. 4).

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