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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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Effector/memory Th2 development and germinal center formation are inhibited by regulatory lymphocytes. (A) Cytokine production by OVA-specific T cells after immunization. Spleen cells from 17/9 DO11.10 RAG−/− immunized mice (No transfer) or 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.) were analyzed for the production of IL-4 and IFN-γ by intracellular staining. Cells were stimulated in vitro with OVA peptide and processed as described in Materials and Methods. The graphic shows anti–IL-4 and anti–IFN-γ plots of KJ1–26+CD4+ gated spleen cells. Unimmunized mice display less than 0.1% of KJ1–26+CD4+ positive cells with either anti–IL-4 or anti–IFN-γ antibodies and no detectable population of IL-4+IFN-γ+ double-positive cells. (B) CD44/CD45RB staining of KJ1–26+CD4+ gated spleen cells from 17/9 DO11.10 RAG−/− immunized mice (No transfer) or 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.). Unimmunized controls analyzed simultaneously as day 14 samples are also shown. (C) Decreased number of PNA+ cells in 17/9D011.10 RAG−/− mice transferred with normal splenocytes. PNA/HA staining is shown for B220+ gated splenocytes from 17/9 DO11.10 RAG−/− mice (No transfer) and from 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells (Tr. Normal Spl.). Unimmunized controls analyzed simultaneously with the 10-d immunization samples are also shown. As expected, germinal center B cells showed downregulation of the B cell receptor.
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fig5: Effector/memory Th2 development and germinal center formation are inhibited by regulatory lymphocytes. (A) Cytokine production by OVA-specific T cells after immunization. Spleen cells from 17/9 DO11.10 RAG−/− immunized mice (No transfer) or 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.) were analyzed for the production of IL-4 and IFN-γ by intracellular staining. Cells were stimulated in vitro with OVA peptide and processed as described in Materials and Methods. The graphic shows anti–IL-4 and anti–IFN-γ plots of KJ1–26+CD4+ gated spleen cells. Unimmunized mice display less than 0.1% of KJ1–26+CD4+ positive cells with either anti–IL-4 or anti–IFN-γ antibodies and no detectable population of IL-4+IFN-γ+ double-positive cells. (B) CD44/CD45RB staining of KJ1–26+CD4+ gated spleen cells from 17/9 DO11.10 RAG−/− immunized mice (No transfer) or 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.). Unimmunized controls analyzed simultaneously as day 14 samples are also shown. (C) Decreased number of PNA+ cells in 17/9D011.10 RAG−/− mice transferred with normal splenocytes. PNA/HA staining is shown for B220+ gated splenocytes from 17/9 DO11.10 RAG−/− mice (No transfer) and from 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells (Tr. Normal Spl.). Unimmunized controls analyzed simultaneously with the 10-d immunization samples are also shown. As expected, germinal center B cells showed downregulation of the B cell receptor.

Mentions: We next investigated whether the formation of effector/memory T helper cells was affected by regulatory lymphocytes. As effector/memory T helper cells are characterized by a fast and intense production of cytokines after stimulation (37), we analyzed the production of IL-4 and IFN-γ by OVA-specific T cells from immunized 17/9 DO11.10 mice ex vivo by intracellular staining. As shown in Fig. 5 A, IL-4+IFN-γ− T cells were virtually undetectable in 17/9 DO11.10 RAG−/− unimmunized mice, were present in low numbers 3 d after immunization and reached up to 10% of the OVA-specific (KJ1–26+) CD4+ splenic cells by day 10 after immunization. IFN-γ+IL-4− cells also increased but to a lesser degree (Fig. 5 A). Strikingly, cells from 17/9 DO11.10 RAG−/− mice transferred with normal splenocytes never developed into high cytokine producers, indicating that the mechanism of regulation does not involve immune deviation to a Th1 phenotype. The intracellular staining data was confirmed by ELISA as well as quantitative competitive RT-PCR (online supplemental Figure S6).


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)

Effector/memory Th2 development and germinal center formation are inhibited by regulatory lymphocytes. (A) Cytokine production by OVA-specific T cells after immunization. Spleen cells from 17/9 DO11.10 RAG−/− immunized mice (No transfer) or 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.) were analyzed for the production of IL-4 and IFN-γ by intracellular staining. Cells were stimulated in vitro with OVA peptide and processed as described in Materials and Methods. The graphic shows anti–IL-4 and anti–IFN-γ plots of KJ1–26+CD4+ gated spleen cells. Unimmunized mice display less than 0.1% of KJ1–26+CD4+ positive cells with either anti–IL-4 or anti–IFN-γ antibodies and no detectable population of IL-4+IFN-γ+ double-positive cells. (B) CD44/CD45RB staining of KJ1–26+CD4+ gated spleen cells from 17/9 DO11.10 RAG−/− immunized mice (No transfer) or 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.). Unimmunized controls analyzed simultaneously as day 14 samples are also shown. (C) Decreased number of PNA+ cells in 17/9D011.10 RAG−/− mice transferred with normal splenocytes. PNA/HA staining is shown for B220+ gated splenocytes from 17/9 DO11.10 RAG−/− mice (No transfer) and from 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells (Tr. Normal Spl.). Unimmunized controls analyzed simultaneously with the 10-d immunization samples are also shown. As expected, germinal center B cells showed downregulation of the B cell receptor.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195981&req=5

fig5: Effector/memory Th2 development and germinal center formation are inhibited by regulatory lymphocytes. (A) Cytokine production by OVA-specific T cells after immunization. Spleen cells from 17/9 DO11.10 RAG−/− immunized mice (No transfer) or 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.) were analyzed for the production of IL-4 and IFN-γ by intracellular staining. Cells were stimulated in vitro with OVA peptide and processed as described in Materials and Methods. The graphic shows anti–IL-4 and anti–IFN-γ plots of KJ1–26+CD4+ gated spleen cells. Unimmunized mice display less than 0.1% of KJ1–26+CD4+ positive cells with either anti–IL-4 or anti–IFN-γ antibodies and no detectable population of IL-4+IFN-γ+ double-positive cells. (B) CD44/CD45RB staining of KJ1–26+CD4+ gated spleen cells from 17/9 DO11.10 RAG−/− immunized mice (No transfer) or 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.). Unimmunized controls analyzed simultaneously as day 14 samples are also shown. (C) Decreased number of PNA+ cells in 17/9D011.10 RAG−/− mice transferred with normal splenocytes. PNA/HA staining is shown for B220+ gated splenocytes from 17/9 DO11.10 RAG−/− mice (No transfer) and from 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells (Tr. Normal Spl.). Unimmunized controls analyzed simultaneously with the 10-d immunization samples are also shown. As expected, germinal center B cells showed downregulation of the B cell receptor.
Mentions: We next investigated whether the formation of effector/memory T helper cells was affected by regulatory lymphocytes. As effector/memory T helper cells are characterized by a fast and intense production of cytokines after stimulation (37), we analyzed the production of IL-4 and IFN-γ by OVA-specific T cells from immunized 17/9 DO11.10 mice ex vivo by intracellular staining. As shown in Fig. 5 A, IL-4+IFN-γ− T cells were virtually undetectable in 17/9 DO11.10 RAG−/− unimmunized mice, were present in low numbers 3 d after immunization and reached up to 10% of the OVA-specific (KJ1–26+) CD4+ splenic cells by day 10 after immunization. IFN-γ+IL-4− cells also increased but to a lesser degree (Fig. 5 A). Strikingly, cells from 17/9 DO11.10 RAG−/− mice transferred with normal splenocytes never developed into high cytokine producers, indicating that the mechanism of regulation does not involve immune deviation to a Th1 phenotype. The intracellular staining data was confirmed by ELISA as well as quantitative competitive RT-PCR (online supplemental Figure S6).

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