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Constitutive expression of interleukin (IL)-4 in vivo causes autoimmune-type disorders in mice.

Erb KJ, Rüger B, von Brevern M, Ryffel B, Schimpl A, Rivett K - J. Exp. Med. (1997)

Bottom Line: The transgenic (tg) expression of interleukin (IL)-4 under the control of a major histocompatibility complex (MHC) class I promoter leads to B cell hyperactivity in mice, characterized by increased B cell surface MHC class II and CD23 expression, elevated responsiveness of the B cells to polyclonal ex vivo stimulation, and increased immunoglobulin (Ig)G1 and IgE serum levels.Therefore the most likely explanation for the increased production of autoantibodies and the autoimmunelike disorders is that IL-4 acts directly on autoreactive B cells by expanding them in a polyclonal manner.Taken together our results show that inappropriate multi-organ expression of IL-4 in vivo leads to autoimmune-type disease in mice.

View Article: PubMed Central - PubMed

Affiliation: The Malaghan Institute of Medical Research, Department of Medicine, Wellington School of Medicine, New Zealand.

ABSTRACT
The transgenic (tg) expression of interleukin (IL)-4 under the control of a major histocompatibility complex (MHC) class I promoter leads to B cell hyperactivity in mice, characterized by increased B cell surface MHC class II and CD23 expression, elevated responsiveness of the B cells to polyclonal ex vivo stimulation, and increased immunoglobulin (Ig)G1 and IgE serum levels. Tg mice develop anemia, glomerulonephritis with complement and immune deposition in the glomeruli, and show increased production of autoantibodies. Treatment of IL-4 tg mice with anti-IL-4 neutralizing antibodies protected the mice from disease development, showing that IL-4 was responsible for the observed disorders. Deletion of superantigen responsive autoreactive T cells in the IL-4 tg mice was normal and treatment of mutant mice with deleting anti-CD4 antibodies failed to ablate the onset of autoimmune-like disease, suggesting that CD4+ T cells were not the primary cause of the disorders. Furthermore, the deletion of B cells reacting against MHC class I molecules was also normal in the IL-4 tg mice. Therefore the most likely explanation for the increased production of autoantibodies and the autoimmunelike disorders is that IL-4 acts directly on autoreactive B cells by expanding them in a polyclonal manner. Taken together our results show that inappropriate multi-organ expression of IL-4 in vivo leads to autoimmune-type disease in mice.

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Hyperreactivity of B cells from the IL-4 tg mice. (A) Upregulation of surface MHC Cl II and CD23 expression and enlargement of tg  B cells. Spleen cells were isolated and then stained with antibodies against  B220, MHC CL II, and CD23. Experiments were repeated six times with  similar results. (B) [3H]Thymidine uptake from B cells from the spleen of  IL-4 tg and littermate control mice stimulated with LPS and anti-μ  F(ab′)2 antibodies. 2 × 105B cells per well were incubated with medium,  LPS (10 μg/ml) and anti-μ F(ab′)2 antibodies (10 μg/ml) in the absence  or presence of 10 μg/ml anti-IL-4–neutralizing antibody (11B11) for 40 h,  pulsed for the last 16 h of the culture period, and then harvested. Mean  [3H]thymidine uptake of triplicates and standard deviations are shown  representative of three separate experiments. (C) B cells from the bone  marrow of IL-4 tg and control mice stimulated with CD40 ligand transfected L cells. 1 × 105B cells from the bone marrow of IL-4 tg and control  mice were incubated with 3 × 104 CD40 ligand–transfected and –untransfected L929 fibroblasts (mitomycin D treated) in the absence or presence  of 10 μg/ml anti-IL-4 neutralizing antibody (11B11). The proliferation  assay was performed as described above.
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Figure 5: Hyperreactivity of B cells from the IL-4 tg mice. (A) Upregulation of surface MHC Cl II and CD23 expression and enlargement of tg B cells. Spleen cells were isolated and then stained with antibodies against B220, MHC CL II, and CD23. Experiments were repeated six times with similar results. (B) [3H]Thymidine uptake from B cells from the spleen of IL-4 tg and littermate control mice stimulated with LPS and anti-μ F(ab′)2 antibodies. 2 × 105B cells per well were incubated with medium, LPS (10 μg/ml) and anti-μ F(ab′)2 antibodies (10 μg/ml) in the absence or presence of 10 μg/ml anti-IL-4–neutralizing antibody (11B11) for 40 h, pulsed for the last 16 h of the culture period, and then harvested. Mean [3H]thymidine uptake of triplicates and standard deviations are shown representative of three separate experiments. (C) B cells from the bone marrow of IL-4 tg and control mice stimulated with CD40 ligand transfected L cells. 1 × 105B cells from the bone marrow of IL-4 tg and control mice were incubated with 3 × 104 CD40 ligand–transfected and –untransfected L929 fibroblasts (mitomycin D treated) in the absence or presence of 10 μg/ml anti-IL-4 neutralizing antibody (11B11). The proliferation assay was performed as described above.

Mentions: Autoantibody production is often associated with polyclonal B cell activation (22, 23). B cells from the spleen, lymph node and bone marrow of IL-4 tg mice express high levels of MHC class II and increased levels of CD23, indicating polyclonal in vivo activation (Fig. 5 A). To investigate whether the observed phenotype also correlates with a greater responsiveness to B cell activators, we stimulated B cells from lymph nodes with LPS and antiμF(ab′)2 antibodies. Fig. 5 B shows that B cells from IL-4 tg mice proliferated more strongly in response to LPS and anti-μF(ab′)2stimulationthan did B cells from littermate controls. Furthermore, splenic B cells from tg mice still showed proliferation to anti-μF(ab′)2at concentrations where normal B cells were no longer reactive (16). To assess whether IL-4 also had a costimulatory effect on B cells activated through the interaction of CD40 with CD40 ligand, we isolated B cells from the bone marrow and activated them with CD40 ligand–transfected L929 fibroblasts (20). Again, B cells from IL-4 tg mice responded more vigorously than did B cells from controls (Fig. 5 C). The addition of anti-IL-4 neutralizing antibodies to the B cell cultures (Fig. 5, B and C) from the IL-4 tg mice had no negative effect on the proliferation of the B cells. This shows that the B cells from the IL-4 tg mice have been activated in vivo and not by tg-derived IL-4 in vitro. Taken together these results, and previously published data (16), support the costimulatory effect of tg IL-4 expression on B cells in vivo leading to a constitutive activation of B cells in the IL-4 tg mice. Interestingly, this polyclonal B cell activation did not lead to a strong increase in total Ig levels in the serum of tg mice, where the average amount of total Ig was only about two- to threefold increased (data not shown). However, IgG1 and especially IgE levels were elevated (16).


Constitutive expression of interleukin (IL)-4 in vivo causes autoimmune-type disorders in mice.

Erb KJ, Rüger B, von Brevern M, Ryffel B, Schimpl A, Rivett K - J. Exp. Med. (1997)

Hyperreactivity of B cells from the IL-4 tg mice. (A) Upregulation of surface MHC Cl II and CD23 expression and enlargement of tg  B cells. Spleen cells were isolated and then stained with antibodies against  B220, MHC CL II, and CD23. Experiments were repeated six times with  similar results. (B) [3H]Thymidine uptake from B cells from the spleen of  IL-4 tg and littermate control mice stimulated with LPS and anti-μ  F(ab′)2 antibodies. 2 × 105B cells per well were incubated with medium,  LPS (10 μg/ml) and anti-μ F(ab′)2 antibodies (10 μg/ml) in the absence  or presence of 10 μg/ml anti-IL-4–neutralizing antibody (11B11) for 40 h,  pulsed for the last 16 h of the culture period, and then harvested. Mean  [3H]thymidine uptake of triplicates and standard deviations are shown  representative of three separate experiments. (C) B cells from the bone  marrow of IL-4 tg and control mice stimulated with CD40 ligand transfected L cells. 1 × 105B cells from the bone marrow of IL-4 tg and control  mice were incubated with 3 × 104 CD40 ligand–transfected and –untransfected L929 fibroblasts (mitomycin D treated) in the absence or presence  of 10 μg/ml anti-IL-4 neutralizing antibody (11B11). The proliferation  assay was performed as described above.
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Related In: Results  -  Collection

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Figure 5: Hyperreactivity of B cells from the IL-4 tg mice. (A) Upregulation of surface MHC Cl II and CD23 expression and enlargement of tg B cells. Spleen cells were isolated and then stained with antibodies against B220, MHC CL II, and CD23. Experiments were repeated six times with similar results. (B) [3H]Thymidine uptake from B cells from the spleen of IL-4 tg and littermate control mice stimulated with LPS and anti-μ F(ab′)2 antibodies. 2 × 105B cells per well were incubated with medium, LPS (10 μg/ml) and anti-μ F(ab′)2 antibodies (10 μg/ml) in the absence or presence of 10 μg/ml anti-IL-4–neutralizing antibody (11B11) for 40 h, pulsed for the last 16 h of the culture period, and then harvested. Mean [3H]thymidine uptake of triplicates and standard deviations are shown representative of three separate experiments. (C) B cells from the bone marrow of IL-4 tg and control mice stimulated with CD40 ligand transfected L cells. 1 × 105B cells from the bone marrow of IL-4 tg and control mice were incubated with 3 × 104 CD40 ligand–transfected and –untransfected L929 fibroblasts (mitomycin D treated) in the absence or presence of 10 μg/ml anti-IL-4 neutralizing antibody (11B11). The proliferation assay was performed as described above.
Mentions: Autoantibody production is often associated with polyclonal B cell activation (22, 23). B cells from the spleen, lymph node and bone marrow of IL-4 tg mice express high levels of MHC class II and increased levels of CD23, indicating polyclonal in vivo activation (Fig. 5 A). To investigate whether the observed phenotype also correlates with a greater responsiveness to B cell activators, we stimulated B cells from lymph nodes with LPS and antiμF(ab′)2 antibodies. Fig. 5 B shows that B cells from IL-4 tg mice proliferated more strongly in response to LPS and anti-μF(ab′)2stimulationthan did B cells from littermate controls. Furthermore, splenic B cells from tg mice still showed proliferation to anti-μF(ab′)2at concentrations where normal B cells were no longer reactive (16). To assess whether IL-4 also had a costimulatory effect on B cells activated through the interaction of CD40 with CD40 ligand, we isolated B cells from the bone marrow and activated them with CD40 ligand–transfected L929 fibroblasts (20). Again, B cells from IL-4 tg mice responded more vigorously than did B cells from controls (Fig. 5 C). The addition of anti-IL-4 neutralizing antibodies to the B cell cultures (Fig. 5, B and C) from the IL-4 tg mice had no negative effect on the proliferation of the B cells. This shows that the B cells from the IL-4 tg mice have been activated in vivo and not by tg-derived IL-4 in vitro. Taken together these results, and previously published data (16), support the costimulatory effect of tg IL-4 expression on B cells in vivo leading to a constitutive activation of B cells in the IL-4 tg mice. Interestingly, this polyclonal B cell activation did not lead to a strong increase in total Ig levels in the serum of tg mice, where the average amount of total Ig was only about two- to threefold increased (data not shown). However, IgG1 and especially IgE levels were elevated (16).

Bottom Line: The transgenic (tg) expression of interleukin (IL)-4 under the control of a major histocompatibility complex (MHC) class I promoter leads to B cell hyperactivity in mice, characterized by increased B cell surface MHC class II and CD23 expression, elevated responsiveness of the B cells to polyclonal ex vivo stimulation, and increased immunoglobulin (Ig)G1 and IgE serum levels.Therefore the most likely explanation for the increased production of autoantibodies and the autoimmunelike disorders is that IL-4 acts directly on autoreactive B cells by expanding them in a polyclonal manner.Taken together our results show that inappropriate multi-organ expression of IL-4 in vivo leads to autoimmune-type disease in mice.

View Article: PubMed Central - PubMed

Affiliation: The Malaghan Institute of Medical Research, Department of Medicine, Wellington School of Medicine, New Zealand.

ABSTRACT
The transgenic (tg) expression of interleukin (IL)-4 under the control of a major histocompatibility complex (MHC) class I promoter leads to B cell hyperactivity in mice, characterized by increased B cell surface MHC class II and CD23 expression, elevated responsiveness of the B cells to polyclonal ex vivo stimulation, and increased immunoglobulin (Ig)G1 and IgE serum levels. Tg mice develop anemia, glomerulonephritis with complement and immune deposition in the glomeruli, and show increased production of autoantibodies. Treatment of IL-4 tg mice with anti-IL-4 neutralizing antibodies protected the mice from disease development, showing that IL-4 was responsible for the observed disorders. Deletion of superantigen responsive autoreactive T cells in the IL-4 tg mice was normal and treatment of mutant mice with deleting anti-CD4 antibodies failed to ablate the onset of autoimmune-like disease, suggesting that CD4+ T cells were not the primary cause of the disorders. Furthermore, the deletion of B cells reacting against MHC class I molecules was also normal in the IL-4 tg mice. Therefore the most likely explanation for the increased production of autoantibodies and the autoimmunelike disorders is that IL-4 acts directly on autoreactive B cells by expanding them in a polyclonal manner. Taken together our results show that inappropriate multi-organ expression of IL-4 in vivo leads to autoimmune-type disease in mice.

Show MeSH
Related in: MedlinePlus