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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

Analysis of activation of OVA-specific T cells. (A) Variation in KJ1–26+CD4+ lymphocyte number in spleens of 17/9 DO11.10 immunized mice. Mean and SD of groups of three mice per time-point are shown. Filled diamonds: 17/9 DO11.10 RAG−/− mice; filled triangles: 17/9 DO11.10 RAG−/− mice transferred with 2 × 107 normal splenocytes; filled squares: 17/9 DO11.10 RAG+ mice. (B) CD25 expression in KJ1–26+CD4+ lymphocytes is not affected by regulatory lymphocytes. FACS® histogram profiles of KJ1–26+CD4+ gated spleen cells from 17/9 DO11.10 RAG−/− mice (No transfer) and 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.). (C) Analysis of cell division of OVA-specific CD4+ lymphocytes. Spleen cells from DO11.10 RAG−/− mice (which contain monoclonal anti-OVA T cells) were labeled with CFSE and transferred (106 OVA-specific T cells) alone or together with unlabeled 2 × 107 normal splenocytes, into 17/9 DO11.10 RAG−/− mice. Recipient mice were immunized 1 d after transfer. Cell division of transferred cells in the spleen was analyzed at several times after immunization. The top three panels illustrate the gating of KJ1–26+CD4+CFSE+ cells and the determination of CFSE intensity intervals defined by CFSE fluorescence peaks (1 to 6, with peak 6 representing T cells that did not divide). The bottom three panels show the distribution of KJ1–26+CD4+ CFSE+ cells (as percentage of total KJ1–26+CD4+ CFSE+) into CFSE fluorescence intervals (numbers 1 to 6 in top panel). Filled diamonds: 17/9 DO11.10 RAG−/− mice; filled triangles: 17/9 DO11.10 RAG−/− mice transferred with 2 × 107 normal splenocytes.
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fig4: Analysis of activation of OVA-specific T cells. (A) Variation in KJ1–26+CD4+ lymphocyte number in spleens of 17/9 DO11.10 immunized mice. Mean and SD of groups of three mice per time-point are shown. Filled diamonds: 17/9 DO11.10 RAG−/− mice; filled triangles: 17/9 DO11.10 RAG−/− mice transferred with 2 × 107 normal splenocytes; filled squares: 17/9 DO11.10 RAG+ mice. (B) CD25 expression in KJ1–26+CD4+ lymphocytes is not affected by regulatory lymphocytes. FACS® histogram profiles of KJ1–26+CD4+ gated spleen cells from 17/9 DO11.10 RAG−/− mice (No transfer) and 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.). (C) Analysis of cell division of OVA-specific CD4+ lymphocytes. Spleen cells from DO11.10 RAG−/− mice (which contain monoclonal anti-OVA T cells) were labeled with CFSE and transferred (106 OVA-specific T cells) alone or together with unlabeled 2 × 107 normal splenocytes, into 17/9 DO11.10 RAG−/− mice. Recipient mice were immunized 1 d after transfer. Cell division of transferred cells in the spleen was analyzed at several times after immunization. The top three panels illustrate the gating of KJ1–26+CD4+CFSE+ cells and the determination of CFSE intensity intervals defined by CFSE fluorescence peaks (1 to 6, with peak 6 representing T cells that did not divide). The bottom three panels show the distribution of KJ1–26+CD4+ CFSE+ cells (as percentage of total KJ1–26+CD4+ CFSE+) into CFSE fluorescence intervals (numbers 1 to 6 in top panel). Filled diamonds: 17/9 DO11.10 RAG−/− mice; filled triangles: 17/9 DO11.10 RAG−/− mice transferred with 2 × 107 normal splenocytes.

Mentions: Early T cell activation and proliferation was assessed by the counting of OVA-specific T cells, the determination of cell size, the expression of early activation markers such as CD25 and CD69, and the rate of cell division. Fig. 4 A shows a kinetic analysis of the numbers of splenic OVA-specific (KJ1–26+CD4+) T cells in immunized 17/9 DO11.10 RAG−/− mice, 17/9 DO11.10 RAG+ and 17/9 DO11.10 RAG−/− mice transferred with normal splenocytes (cell numbers of unimmunized mice shown at time point zero). Expansion of the anti-OVA T cells was similar in the three groups, with cell numbers peaking around days 2 and 3 d after immunization and declining thereafter. Likewise, the kinetics of upregulation and downregulation of CD25 and CD69 expression in spleen cells from 17/9 DO11.10 RAG−/− mice and 17/9 DO11.10 RAG−/− mice transferred with normal splenocytes showed no differences. Most OVA-specific T cells upregulated CD25 and CD69 expression levels at 15 h after immunization in both groups (Fig. 4 B, and online supplemental Figure S4). Similar kinetics of expansion/contraction in the number of OVA-specific T cells and expression of activation markers were observed after analysis of lymph node cells (online supplemental Figure S5).


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)

Analysis of activation of OVA-specific T cells. (A) Variation in KJ1–26+CD4+ lymphocyte number in spleens of 17/9 DO11.10 immunized mice. Mean and SD of groups of three mice per time-point are shown. Filled diamonds: 17/9 DO11.10 RAG−/− mice; filled triangles: 17/9 DO11.10 RAG−/− mice transferred with 2 × 107 normal splenocytes; filled squares: 17/9 DO11.10 RAG+ mice. (B) CD25 expression in KJ1–26+CD4+ lymphocytes is not affected by regulatory lymphocytes. FACS® histogram profiles of KJ1–26+CD4+ gated spleen cells from 17/9 DO11.10 RAG−/− mice (No transfer) and 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.). (C) Analysis of cell division of OVA-specific CD4+ lymphocytes. Spleen cells from DO11.10 RAG−/− mice (which contain monoclonal anti-OVA T cells) were labeled with CFSE and transferred (106 OVA-specific T cells) alone or together with unlabeled 2 × 107 normal splenocytes, into 17/9 DO11.10 RAG−/− mice. Recipient mice were immunized 1 d after transfer. Cell division of transferred cells in the spleen was analyzed at several times after immunization. The top three panels illustrate the gating of KJ1–26+CD4+CFSE+ cells and the determination of CFSE intensity intervals defined by CFSE fluorescence peaks (1 to 6, with peak 6 representing T cells that did not divide). The bottom three panels show the distribution of KJ1–26+CD4+ CFSE+ cells (as percentage of total KJ1–26+CD4+ CFSE+) into CFSE fluorescence intervals (numbers 1 to 6 in top panel). Filled diamonds: 17/9 DO11.10 RAG−/− mice; filled triangles: 17/9 DO11.10 RAG−/− mice transferred with 2 × 107 normal splenocytes.
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Related In: Results  -  Collection

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fig4: Analysis of activation of OVA-specific T cells. (A) Variation in KJ1–26+CD4+ lymphocyte number in spleens of 17/9 DO11.10 immunized mice. Mean and SD of groups of three mice per time-point are shown. Filled diamonds: 17/9 DO11.10 RAG−/− mice; filled triangles: 17/9 DO11.10 RAG−/− mice transferred with 2 × 107 normal splenocytes; filled squares: 17/9 DO11.10 RAG+ mice. (B) CD25 expression in KJ1–26+CD4+ lymphocytes is not affected by regulatory lymphocytes. FACS® histogram profiles of KJ1–26+CD4+ gated spleen cells from 17/9 DO11.10 RAG−/− mice (No transfer) and 17/9 DO11.10 RAG−/− mice that were transferred with 2 × 107 normal spleen cells one day before immunization (Tr. Normal Spl.). (C) Analysis of cell division of OVA-specific CD4+ lymphocytes. Spleen cells from DO11.10 RAG−/− mice (which contain monoclonal anti-OVA T cells) were labeled with CFSE and transferred (106 OVA-specific T cells) alone or together with unlabeled 2 × 107 normal splenocytes, into 17/9 DO11.10 RAG−/− mice. Recipient mice were immunized 1 d after transfer. Cell division of transferred cells in the spleen was analyzed at several times after immunization. The top three panels illustrate the gating of KJ1–26+CD4+CFSE+ cells and the determination of CFSE intensity intervals defined by CFSE fluorescence peaks (1 to 6, with peak 6 representing T cells that did not divide). The bottom three panels show the distribution of KJ1–26+CD4+ CFSE+ cells (as percentage of total KJ1–26+CD4+ CFSE+) into CFSE fluorescence intervals (numbers 1 to 6 in top panel). Filled diamonds: 17/9 DO11.10 RAG−/− mice; filled triangles: 17/9 DO11.10 RAG−/− mice transferred with 2 × 107 normal splenocytes.
Mentions: Early T cell activation and proliferation was assessed by the counting of OVA-specific T cells, the determination of cell size, the expression of early activation markers such as CD25 and CD69, and the rate of cell division. Fig. 4 A shows a kinetic analysis of the numbers of splenic OVA-specific (KJ1–26+CD4+) T cells in immunized 17/9 DO11.10 RAG−/− mice, 17/9 DO11.10 RAG+ and 17/9 DO11.10 RAG−/− mice transferred with normal splenocytes (cell numbers of unimmunized mice shown at time point zero). Expansion of the anti-OVA T cells was similar in the three groups, with cell numbers peaking around days 2 and 3 d after immunization and declining thereafter. Likewise, the kinetics of upregulation and downregulation of CD25 and CD69 expression in spleen cells from 17/9 DO11.10 RAG−/− mice and 17/9 DO11.10 RAG−/− mice transferred with normal splenocytes showed no differences. Most OVA-specific T cells upregulated CD25 and CD69 expression levels at 15 h after immunization in both groups (Fig. 4 B, and online supplemental Figure S4). Similar kinetics of expansion/contraction in the number of OVA-specific T cells and expression of activation markers were observed after analysis of lymph node cells (online supplemental Figure S5).

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