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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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Related in: MedlinePlus

Development of hyper IgE can be prevented by normal TCR α/β+CD4+ splenocytes. (A) Total (2.5 × 107) or fractionated spleen cells (CD4-depleted, CD8-depleted, B220-depleted: 2 × 107) from wild-type BALB/c mice or total spleen cells from TCR α−/− mice (2 × 107) were transferred into 17/9 DO11.10 RAG−/− recipients 1 d before immunization. Total IgE serum levels were determined 14 d after immunization. n = 6–8 mice per group. (B) Purified spleen cells populations obtained from wild-type BALB/c mice or BALB/c μMT−/− mice were transferred to 17/9 DO11.10 RAG−/− recipients one day before immunization. Serum IgE levels were determined 19 d after immunization. Exp 1: three cell populations were obtained from wild-type BALB/c donors and transferred: (1) purified CD4+CD25+ spleen cells (6 × 105); (2) spleen cells depleted of B220+, CD8+, and CD25+ lymphocytes (6 × 105 or 3 × 106 CD4+CD25− cells transferred); (3) spleen cells depleted of B220+ and CD8+ lymphocytes (6 × 105 or 3 × 106 total CD4+ T cells transferred). Exp 2: purified CD4+CD25+ spleen cells, and spleen cells depleted of CD25+ cells were obtained from BALB/c μMT−/− donors and transferred in doses of 5 × 105 or 105 CD4+ T cells. Exp 3: CD4+CD25+ and CD4+CD25− donor spleen cells were obtained from BALB/c μMT−/− donors. To purify CD4+CD25− cells, spleen cells were first depleted of CD25+ cells. Subsequently, CD4+ cells were positively sorted. (C) Typical FACS® profiles of purified donor populations are shown. Dot plots contain the profiles of cells in a broad FSC/SSC lymphocyte gate. The three plots on the left show donor cells from Exp 2. The plot on the right shows purified CD4+CD25− cells from Exp 3.
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fig3: Development of hyper IgE can be prevented by normal TCR α/β+CD4+ splenocytes. (A) Total (2.5 × 107) or fractionated spleen cells (CD4-depleted, CD8-depleted, B220-depleted: 2 × 107) from wild-type BALB/c mice or total spleen cells from TCR α−/− mice (2 × 107) were transferred into 17/9 DO11.10 RAG−/− recipients 1 d before immunization. Total IgE serum levels were determined 14 d after immunization. n = 6–8 mice per group. (B) Purified spleen cells populations obtained from wild-type BALB/c mice or BALB/c μMT−/− mice were transferred to 17/9 DO11.10 RAG−/− recipients one day before immunization. Serum IgE levels were determined 19 d after immunization. Exp 1: three cell populations were obtained from wild-type BALB/c donors and transferred: (1) purified CD4+CD25+ spleen cells (6 × 105); (2) spleen cells depleted of B220+, CD8+, and CD25+ lymphocytes (6 × 105 or 3 × 106 CD4+CD25− cells transferred); (3) spleen cells depleted of B220+ and CD8+ lymphocytes (6 × 105 or 3 × 106 total CD4+ T cells transferred). Exp 2: purified CD4+CD25+ spleen cells, and spleen cells depleted of CD25+ cells were obtained from BALB/c μMT−/− donors and transferred in doses of 5 × 105 or 105 CD4+ T cells. Exp 3: CD4+CD25+ and CD4+CD25− donor spleen cells were obtained from BALB/c μMT−/− donors. To purify CD4+CD25− cells, spleen cells were first depleted of CD25+ cells. Subsequently, CD4+ cells were positively sorted. (C) Typical FACS® profiles of purified donor populations are shown. Dot plots contain the profiles of cells in a broad FSC/SSC lymphocyte gate. The three plots on the left show donor cells from Exp 2. The plot on the right shows purified CD4+CD25− cells from Exp 3.

Mentions: The rearranged V(D)J genes of the 17/9 antibody were cloned from the 17/9 hybridoma (26) and used to construct targeting vectors for the heavy and κ chain Ig loci. Targeting constructs containing a neo-loxP cassette were transfected into the J1 embryonic stem (ES) cell line and G418 resistant colonies carrying homologous integrations were identified by Southern analysis. Probes A, B, C, and D in Fig. 1 were used to characterize the 5′ and 3′ targeting sites. The neo-loxP cassette was removed by cre-mediated recombination. Mice carrying both rearranged receptor genes were generated by crosses between single targeted mice, and are referred to as 17/9 mice. All mice were backcrossed onto a BALB/c background and were housed under specific pathogen-free conditions at the Skirball Institute, New York University School of Medicine. All 17/9 DO11.10 RAG−/− animals in experiments that involved transfer of CD4+CD25− splenocytes (Fig. 3 B), were given sulfamethoxazole (400 mg/liter)/trimethoprim (80 mg/liter) in the drinking water for the duration of the experiment. This treatment prevented wasting of mice transferred with CD4+CD25− cells.


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)

Development of hyper IgE can be prevented by normal TCR α/β+CD4+ splenocytes. (A) Total (2.5 × 107) or fractionated spleen cells (CD4-depleted, CD8-depleted, B220-depleted: 2 × 107) from wild-type BALB/c mice or total spleen cells from TCR α−/− mice (2 × 107) were transferred into 17/9 DO11.10 RAG−/− recipients 1 d before immunization. Total IgE serum levels were determined 14 d after immunization. n = 6–8 mice per group. (B) Purified spleen cells populations obtained from wild-type BALB/c mice or BALB/c μMT−/− mice were transferred to 17/9 DO11.10 RAG−/− recipients one day before immunization. Serum IgE levels were determined 19 d after immunization. Exp 1: three cell populations were obtained from wild-type BALB/c donors and transferred: (1) purified CD4+CD25+ spleen cells (6 × 105); (2) spleen cells depleted of B220+, CD8+, and CD25+ lymphocytes (6 × 105 or 3 × 106 CD4+CD25− cells transferred); (3) spleen cells depleted of B220+ and CD8+ lymphocytes (6 × 105 or 3 × 106 total CD4+ T cells transferred). Exp 2: purified CD4+CD25+ spleen cells, and spleen cells depleted of CD25+ cells were obtained from BALB/c μMT−/− donors and transferred in doses of 5 × 105 or 105 CD4+ T cells. Exp 3: CD4+CD25+ and CD4+CD25− donor spleen cells were obtained from BALB/c μMT−/− donors. To purify CD4+CD25− cells, spleen cells were first depleted of CD25+ cells. Subsequently, CD4+ cells were positively sorted. (C) Typical FACS® profiles of purified donor populations are shown. Dot plots contain the profiles of cells in a broad FSC/SSC lymphocyte gate. The three plots on the left show donor cells from Exp 2. The plot on the right shows purified CD4+CD25− cells from Exp 3.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2195981&req=5

fig3: Development of hyper IgE can be prevented by normal TCR α/β+CD4+ splenocytes. (A) Total (2.5 × 107) or fractionated spleen cells (CD4-depleted, CD8-depleted, B220-depleted: 2 × 107) from wild-type BALB/c mice or total spleen cells from TCR α−/− mice (2 × 107) were transferred into 17/9 DO11.10 RAG−/− recipients 1 d before immunization. Total IgE serum levels were determined 14 d after immunization. n = 6–8 mice per group. (B) Purified spleen cells populations obtained from wild-type BALB/c mice or BALB/c μMT−/− mice were transferred to 17/9 DO11.10 RAG−/− recipients one day before immunization. Serum IgE levels were determined 19 d after immunization. Exp 1: three cell populations were obtained from wild-type BALB/c donors and transferred: (1) purified CD4+CD25+ spleen cells (6 × 105); (2) spleen cells depleted of B220+, CD8+, and CD25+ lymphocytes (6 × 105 or 3 × 106 CD4+CD25− cells transferred); (3) spleen cells depleted of B220+ and CD8+ lymphocytes (6 × 105 or 3 × 106 total CD4+ T cells transferred). Exp 2: purified CD4+CD25+ spleen cells, and spleen cells depleted of CD25+ cells were obtained from BALB/c μMT−/− donors and transferred in doses of 5 × 105 or 105 CD4+ T cells. Exp 3: CD4+CD25+ and CD4+CD25− donor spleen cells were obtained from BALB/c μMT−/− donors. To purify CD4+CD25− cells, spleen cells were first depleted of CD25+ cells. Subsequently, CD4+ cells were positively sorted. (C) Typical FACS® profiles of purified donor populations are shown. Dot plots contain the profiles of cells in a broad FSC/SSC lymphocyte gate. The three plots on the left show donor cells from Exp 2. The plot on the right shows purified CD4+CD25− cells from Exp 3.
Mentions: The rearranged V(D)J genes of the 17/9 antibody were cloned from the 17/9 hybridoma (26) and used to construct targeting vectors for the heavy and κ chain Ig loci. Targeting constructs containing a neo-loxP cassette were transfected into the J1 embryonic stem (ES) cell line and G418 resistant colonies carrying homologous integrations were identified by Southern analysis. Probes A, B, C, and D in Fig. 1 were used to characterize the 5′ and 3′ targeting sites. The neo-loxP cassette was removed by cre-mediated recombination. Mice carrying both rearranged receptor genes were generated by crosses between single targeted mice, and are referred to as 17/9 mice. All mice were backcrossed onto a BALB/c background and were housed under specific pathogen-free conditions at the Skirball Institute, New York University School of Medicine. All 17/9 DO11.10 RAG−/− animals in experiments that involved transfer of CD4+CD25− splenocytes (Fig. 3 B), were given sulfamethoxazole (400 mg/liter)/trimethoprim (80 mg/liter) in the drinking water for the duration of the experiment. This treatment prevented wasting of mice transferred with CD4+CD25− cells.

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