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Hyper immunoglobulin E response in mice with monoclonal populations of B and T lymphocytes.

Curotto de Lafaille MA, Muriglan S, Sunshine MJ, Lei Y, Kutchukhidze N, Furtado GC, Wensky AK, Olivares-Villagómez D, Lafaille JJ - J. Exp. Med. (2001)

Bottom Line: This unusually high IgE response was prevented by the infusion of regulatory alpha/beta CD4(+) T cells belonging to both CD25(+) and CD25(-) subpopulations.The regulation by the infused T cells impeded the development of fully competent OVA-specific effector/memory Th2 lymphocytes without inhibiting the initial proliferative response of T cells or promoting activation-induced cell death.Our results indicate that hyper IgE responses do not occur in normal individuals due to the presence of regulatory T cells, and imply that the induction of regulatory CD4(+) T cells could be used for the prevention of atopy.

View Article: PubMed Central - PubMed

Affiliation: Program of Molecular Pathogenesis, Skirball Institute for Biomolecular Medicine, New York University School of Medicine, New York, NY 10016, USA.

ABSTRACT
A key event in the pathogenesis of allergies is the production of antibodies of the immunoglobulin (Ig)E class. In normal individuals the levels of IgE are tightly regulated, as illustrated by the low serum IgE concentration. In addition, multiple immunizations are usually required to generate detectable IgE responses in normal experimental animals. To define the parameters that regulate IgE production in vivo, we generated mice bearing monoclonal populations of B and T lymphocytes specific for influenza virus hemagglutinin (HA) and chicken ovalbumin (OVA), respectively. A single immunization of the monoclonal mice with the cross-linked OVA-HA antigen led to serum IgE levels that reached 30-200 microg/ml. This unusually high IgE response was prevented by the infusion of regulatory alpha/beta CD4(+) T cells belonging to both CD25(+) and CD25(-) subpopulations. The regulation by the infused T cells impeded the development of fully competent OVA-specific effector/memory Th2 lymphocytes without inhibiting the initial proliferative response of T cells or promoting activation-induced cell death. Our results indicate that hyper IgE responses do not occur in normal individuals due to the presence of regulatory T cells, and imply that the induction of regulatory CD4(+) T cells could be used for the prevention of atopy.

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Hyper IgE production in mice with monoclonal populations of B and T lymphocytes. (A) HA-specific B cells and OVA-specific (KJ1–26+) T cells in 17/9 DO11.10 RAG−/− mice (plots on left) and 17/9 DO11.10 RAG+ mice (plots on right). FACS® analysis of peripheral blood lymphocytes from 4-wk-old mice. Only 10% of the CD3+ KJ1–26− T cells are CD8+, comprising less than 2% of the FSC × SSC lymphocyte gate. (B) Total serum IgE levels 14 d after single immunization. n = 7 mice per group. (C) Ratios of HA-specific serum immunoglobulin titers in sera of 17/9 DO11.10 RAG−/− and 17/9 DO11.10 RAG+ mice 14 d after single immunization. n = 7 mice per group. (D) T cells determine the hyper IgE phenotype. 5 × 106 spleen cells from 17/9 RAG−/− mice (containing monoclonal anti-HA B cells) were transferred into DO11.10 RAG−/− mice (left bar), DO11.10 μMT−/− mice (middle bar), and DO11.10 RAG+μMT+ mice (right bar) 3 d after single immunization. The graphic shows total IgE levels in sera of mice on day 14 after immunization. n = 3 to 4 mice per group.
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fig2: Hyper IgE production in mice with monoclonal populations of B and T lymphocytes. (A) HA-specific B cells and OVA-specific (KJ1–26+) T cells in 17/9 DO11.10 RAG−/− mice (plots on left) and 17/9 DO11.10 RAG+ mice (plots on right). FACS® analysis of peripheral blood lymphocytes from 4-wk-old mice. Only 10% of the CD3+ KJ1–26− T cells are CD8+, comprising less than 2% of the FSC × SSC lymphocyte gate. (B) Total serum IgE levels 14 d after single immunization. n = 7 mice per group. (C) Ratios of HA-specific serum immunoglobulin titers in sera of 17/9 DO11.10 RAG−/− and 17/9 DO11.10 RAG+ mice 14 d after single immunization. n = 7 mice per group. (D) T cells determine the hyper IgE phenotype. 5 × 106 spleen cells from 17/9 RAG−/− mice (containing monoclonal anti-HA B cells) were transferred into DO11.10 RAG−/− mice (left bar), DO11.10 μMT−/− mice (middle bar), and DO11.10 RAG+μMT+ mice (right bar) 3 d after single immunization. The graphic shows total IgE levels in sera of mice on day 14 after immunization. n = 3 to 4 mice per group.

Mentions: To monitor the induction and regulation of the IgE response we used mice that produce a high frequency of T and B lymphocytes specific for defined antigens. A new mouse strain which carries V(D)J replacements for the heavy and light chain genes of the monoclonal antibody 17/9 was generated by gene targeting (Fig. 1). This mouse strain, referred to as 17/9, harbors a large number of mature B lymphocytes specific for HA of the influenza virus. 17/9 DO11.10 RAG−/− mice were generated by crossing the 17/9 mice with the DO11.10 anti-OVA T cell receptor transgenic mice (27), and subsequently incorporating a RAG1 mutation (28). 17/9 DO11.10 RAG−/− mice produce monoclonal populations of anti-HA B lymphocytes and anti-OVA CD4+ T lymphocytes. In contrast, 17/9 DO11.10 RAG+ mice harbor, in addition to the HA and OVA-specific lymphocytes, other B and T lymphocytes expressing antigen receptors encoded by the endogenous loci (Fig. 2 A). Immunization of the 17/9 DO11.10 RAG−/− and 17/9 DO11.10 RAG+ mice with a cross-linked OVA-HA antigen induced a T cell–dependent antibody response in both groups of mice. However, the outcome of the immunization was very different in both groups, resulting in a two order of magnitude increase in total IgE levels in the 17/9 DO11.10 RAG−/− but not in the 17/9 DO11.10 RAG+ mice (Fig. 2 B). The possibility that the observed differences reflected an impaired clearance of serum Igs in the RAG−/− strain rather than increased class switching was ruled out by the measurement of Ig half-life (online supplemental Figure S1). The production of HA-specific antibodies of all IgG isotypes was stimulated in mice of both strains by the immunization (online supplemental Figure S2). However, serum titers for HA-specific IgGs were higher in the 17/9 DO11.10 RAG−/− strain than in the RAG+ strain. Among HA-specific IgGs, the largest difference was in IgG1, which reached titers ∼20 fold higher in 17/9 DO11.10 RAG−/− mice than in their RAG+ littermates (Fig. 2 C).


Hyper immunoglobulin E response in mice with monoclonal populations of B and T lymphocytes.

Curotto de Lafaille MA, Muriglan S, Sunshine MJ, Lei Y, Kutchukhidze N, Furtado GC, Wensky AK, Olivares-Villagómez D, Lafaille JJ - J. Exp. Med. (2001)

Hyper IgE production in mice with monoclonal populations of B and T lymphocytes. (A) HA-specific B cells and OVA-specific (KJ1–26+) T cells in 17/9 DO11.10 RAG−/− mice (plots on left) and 17/9 DO11.10 RAG+ mice (plots on right). FACS® analysis of peripheral blood lymphocytes from 4-wk-old mice. Only 10% of the CD3+ KJ1–26− T cells are CD8+, comprising less than 2% of the FSC × SSC lymphocyte gate. (B) Total serum IgE levels 14 d after single immunization. n = 7 mice per group. (C) Ratios of HA-specific serum immunoglobulin titers in sera of 17/9 DO11.10 RAG−/− and 17/9 DO11.10 RAG+ mice 14 d after single immunization. n = 7 mice per group. (D) T cells determine the hyper IgE phenotype. 5 × 106 spleen cells from 17/9 RAG−/− mice (containing monoclonal anti-HA B cells) were transferred into DO11.10 RAG−/− mice (left bar), DO11.10 μMT−/− mice (middle bar), and DO11.10 RAG+μMT+ mice (right bar) 3 d after single immunization. The graphic shows total IgE levels in sera of mice on day 14 after immunization. n = 3 to 4 mice per group.
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Related In: Results  -  Collection

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fig2: Hyper IgE production in mice with monoclonal populations of B and T lymphocytes. (A) HA-specific B cells and OVA-specific (KJ1–26+) T cells in 17/9 DO11.10 RAG−/− mice (plots on left) and 17/9 DO11.10 RAG+ mice (plots on right). FACS® analysis of peripheral blood lymphocytes from 4-wk-old mice. Only 10% of the CD3+ KJ1–26− T cells are CD8+, comprising less than 2% of the FSC × SSC lymphocyte gate. (B) Total serum IgE levels 14 d after single immunization. n = 7 mice per group. (C) Ratios of HA-specific serum immunoglobulin titers in sera of 17/9 DO11.10 RAG−/− and 17/9 DO11.10 RAG+ mice 14 d after single immunization. n = 7 mice per group. (D) T cells determine the hyper IgE phenotype. 5 × 106 spleen cells from 17/9 RAG−/− mice (containing monoclonal anti-HA B cells) were transferred into DO11.10 RAG−/− mice (left bar), DO11.10 μMT−/− mice (middle bar), and DO11.10 RAG+μMT+ mice (right bar) 3 d after single immunization. The graphic shows total IgE levels in sera of mice on day 14 after immunization. n = 3 to 4 mice per group.
Mentions: To monitor the induction and regulation of the IgE response we used mice that produce a high frequency of T and B lymphocytes specific for defined antigens. A new mouse strain which carries V(D)J replacements for the heavy and light chain genes of the monoclonal antibody 17/9 was generated by gene targeting (Fig. 1). This mouse strain, referred to as 17/9, harbors a large number of mature B lymphocytes specific for HA of the influenza virus. 17/9 DO11.10 RAG−/− mice were generated by crossing the 17/9 mice with the DO11.10 anti-OVA T cell receptor transgenic mice (27), and subsequently incorporating a RAG1 mutation (28). 17/9 DO11.10 RAG−/− mice produce monoclonal populations of anti-HA B lymphocytes and anti-OVA CD4+ T lymphocytes. In contrast, 17/9 DO11.10 RAG+ mice harbor, in addition to the HA and OVA-specific lymphocytes, other B and T lymphocytes expressing antigen receptors encoded by the endogenous loci (Fig. 2 A). Immunization of the 17/9 DO11.10 RAG−/− and 17/9 DO11.10 RAG+ mice with a cross-linked OVA-HA antigen induced a T cell–dependent antibody response in both groups of mice. However, the outcome of the immunization was very different in both groups, resulting in a two order of magnitude increase in total IgE levels in the 17/9 DO11.10 RAG−/− but not in the 17/9 DO11.10 RAG+ mice (Fig. 2 B). The possibility that the observed differences reflected an impaired clearance of serum Igs in the RAG−/− strain rather than increased class switching was ruled out by the measurement of Ig half-life (online supplemental Figure S1). The production of HA-specific antibodies of all IgG isotypes was stimulated in mice of both strains by the immunization (online supplemental Figure S2). However, serum titers for HA-specific IgGs were higher in the 17/9 DO11.10 RAG−/− strain than in the RAG+ strain. Among HA-specific IgGs, the largest difference was in IgG1, which reached titers ∼20 fold higher in 17/9 DO11.10 RAG−/− mice than in their RAG+ littermates (Fig. 2 C).

Bottom Line: This unusually high IgE response was prevented by the infusion of regulatory alpha/beta CD4(+) T cells belonging to both CD25(+) and CD25(-) subpopulations.The regulation by the infused T cells impeded the development of fully competent OVA-specific effector/memory Th2 lymphocytes without inhibiting the initial proliferative response of T cells or promoting activation-induced cell death.Our results indicate that hyper IgE responses do not occur in normal individuals due to the presence of regulatory T cells, and imply that the induction of regulatory CD4(+) T cells could be used for the prevention of atopy.

View Article: PubMed Central - PubMed

Affiliation: Program of Molecular Pathogenesis, Skirball Institute for Biomolecular Medicine, New York University School of Medicine, New York, NY 10016, USA.

ABSTRACT
A key event in the pathogenesis of allergies is the production of antibodies of the immunoglobulin (Ig)E class. In normal individuals the levels of IgE are tightly regulated, as illustrated by the low serum IgE concentration. In addition, multiple immunizations are usually required to generate detectable IgE responses in normal experimental animals. To define the parameters that regulate IgE production in vivo, we generated mice bearing monoclonal populations of B and T lymphocytes specific for influenza virus hemagglutinin (HA) and chicken ovalbumin (OVA), respectively. A single immunization of the monoclonal mice with the cross-linked OVA-HA antigen led to serum IgE levels that reached 30-200 microg/ml. This unusually high IgE response was prevented by the infusion of regulatory alpha/beta CD4(+) T cells belonging to both CD25(+) and CD25(-) subpopulations. The regulation by the infused T cells impeded the development of fully competent OVA-specific effector/memory Th2 lymphocytes without inhibiting the initial proliferative response of T cells or promoting activation-induced cell death. Our results indicate that hyper IgE responses do not occur in normal individuals due to the presence of regulatory T cells, and imply that the induction of regulatory CD4(+) T cells could be used for the prevention of atopy.

Show MeSH
Related in: MedlinePlus