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Interferon (IFN) consensus sequence-binding protein, a transcription factor of the IFN regulatory factor family, regulates immune responses in vivo through control of interleukin 12 expression.

Giese NA, Gabriele L, Doherty TM, Klinman DM, Tadesse-Heath L, Contursi C, Epstein SL, Morse HC - J. Exp. Med. (1997)

Bottom Line: In sera of normal mice, immunoglobulin (Ig)G2a antibodies were dominant over IgG1 antibodies, a pattern indicative of a T helper cell type 1 (Th1)-driven response.Infected ICSBP-deficient mice developed fulminant, disseminated leishmaniasis as a result of failure to mount a Th1-mediated curative response, although T cells remained capable of secreting IFN-gamma and macrophages of producing nitric oxide.This indicates that ICSBP is a deciding factor in Th responses governing humoral and cellular immunity through its role in regulating IL-12 expression.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunopathology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-0760, USA. ngiese@atlas.niaid.nih.gov

ABSTRACT
Mice with a mutation of the gene encoding interferon consensus sequence-binding protein (ICSBP) develop a chronic myelogenous leukemia-like syndrome and mount impaired responses to certain viral and bacterial infections. To gain a mechanistic understanding of the contributions of ICSBP to humoral and cellular immunity, we characterized the responses of control and ICSBP-/- mice to infection with influenza A (flu) and Leishmania major (L. major). Mice of both genotypes survived infections with flu, but differed markedly in the isotype distribution of antiflu antibodies. In sera of normal mice, immunoglobulin (Ig)G2a antibodies were dominant over IgG1 antibodies, a pattern indicative of a T helper cell type 1 (Th1)-driven response. In sera of ICSBP-/- mice, however, IgG1 antibodies dominated over IgG2a antibodies, a pattern indicative of a Th2-driven response. The dominance of IgG1 and IgE over IgG2a was detected in the sera of uninfected mice as well. A seeming Th2 bias of ICSBP-deficient mice was also uncovered in their inability to control infection with L. major, where resistance is known to be dependent on IL-12 and IFN-gamma as components of a Th1 response. Infected ICSBP-deficient mice developed fulminant, disseminated leishmaniasis as a result of failure to mount a Th1-mediated curative response, although T cells remained capable of secreting IFN-gamma and macrophages of producing nitric oxide. Compromised Th1 differentiation in ICSBP-/- mice could not be attributed to hyporesponsiveness of CD4(+) T cells to interleukin (IL)-12; however, the ability of uninfected and infected ICSBP-deficient mice to produce IL-12 was markedly impaired. This indicates that ICSBP is a deciding factor in Th responses governing humoral and cellular immunity through its role in regulating IL-12 expression.

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Impaired IL-12 expression in ICSBP−/− mice. (A) Concentration of IL-12p40 in supernatants obtained from indicated cell populations after 18–24 h of incubation with 200 U/ml IFN-γ in combination  with 100 ng/ml LPS (left) or 1:1,000 diluted preparation of CD40L (right).  Spleen (spl) and bone marrow (bm) cells were plated at 2 × 106/well; normal peritoneal washout (PC) or thioglycollate-elicited exudate (PEX)  cells were used at 6 × 105/well. In a number of experiments, serial dilution analysis was performed. Data indicate the mean ± SEM of IL-12p40  concentration in supernatants from the indicated cell populations as well  as from macrophages (M∅ ) enriched by adherence. The results summarize data from two to six experiments with two to three mice in each experiment. (B) ELISpot analysis of the ability of ODN or E. coli DNA to  trigger IL-12p40 secretion by cells from spleen (left) or lymph nodes (right)  of ICSBP−/− (KO) and ICSBP+/+ (WT) mice. The frequency of IL-12– producing cells (mean ± SEM) for three individual mice is shown. (C)  RT-PCR analysis of IL-12p40 RNA expression after 6 h of stimulation  of 107 hematopoietic cells with 100 ng/ml LPS ± 100 U/ml IFN-γ. Data  shown are representative of three independent studies. ns, no sample.
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Figure 2: Impaired IL-12 expression in ICSBP−/− mice. (A) Concentration of IL-12p40 in supernatants obtained from indicated cell populations after 18–24 h of incubation with 200 U/ml IFN-γ in combination with 100 ng/ml LPS (left) or 1:1,000 diluted preparation of CD40L (right). Spleen (spl) and bone marrow (bm) cells were plated at 2 × 106/well; normal peritoneal washout (PC) or thioglycollate-elicited exudate (PEX) cells were used at 6 × 105/well. In a number of experiments, serial dilution analysis was performed. Data indicate the mean ± SEM of IL-12p40 concentration in supernatants from the indicated cell populations as well as from macrophages (M∅ ) enriched by adherence. The results summarize data from two to six experiments with two to three mice in each experiment. (B) ELISpot analysis of the ability of ODN or E. coli DNA to trigger IL-12p40 secretion by cells from spleen (left) or lymph nodes (right) of ICSBP−/− (KO) and ICSBP+/+ (WT) mice. The frequency of IL-12– producing cells (mean ± SEM) for three individual mice is shown. (C) RT-PCR analysis of IL-12p40 RNA expression after 6 h of stimulation of 107 hematopoietic cells with 100 ng/ml LPS ± 100 U/ml IFN-γ. Data shown are representative of three independent studies. ns, no sample.

Mentions: Although transcripts of IL-12p35 were comparable in ICSBP−/− and ICSBP+/+ mice, transcripts for IL-12p40 were markedly lower in ICSBP-deficient mice (Fig. 1 B; reference 21). IL-12p35 expression is constitutive in many cell types, whereas IL-12p40 is produced after activation of monocytes, macrophages, neutrophils, dendritic cells, and B cells by various stimuli (for review see reference 41). In the first set of experiments, we examined the ability of cells from spleen, peritoneum, or bone marrow, either unseparated or enriched by adherence for macrophages, to secrete IL-12. The cultures were treated with IFN-γ and either LPS or CD40L (Fig. 2 A). Cells from wild-type mice produced consistently higher levels of IL-12p40 than did cells from ICSBP-deficient mice, with the responses from the latter often being at or below the limits of detection.


Interferon (IFN) consensus sequence-binding protein, a transcription factor of the IFN regulatory factor family, regulates immune responses in vivo through control of interleukin 12 expression.

Giese NA, Gabriele L, Doherty TM, Klinman DM, Tadesse-Heath L, Contursi C, Epstein SL, Morse HC - J. Exp. Med. (1997)

Impaired IL-12 expression in ICSBP−/− mice. (A) Concentration of IL-12p40 in supernatants obtained from indicated cell populations after 18–24 h of incubation with 200 U/ml IFN-γ in combination  with 100 ng/ml LPS (left) or 1:1,000 diluted preparation of CD40L (right).  Spleen (spl) and bone marrow (bm) cells were plated at 2 × 106/well; normal peritoneal washout (PC) or thioglycollate-elicited exudate (PEX)  cells were used at 6 × 105/well. In a number of experiments, serial dilution analysis was performed. Data indicate the mean ± SEM of IL-12p40  concentration in supernatants from the indicated cell populations as well  as from macrophages (M∅ ) enriched by adherence. The results summarize data from two to six experiments with two to three mice in each experiment. (B) ELISpot analysis of the ability of ODN or E. coli DNA to  trigger IL-12p40 secretion by cells from spleen (left) or lymph nodes (right)  of ICSBP−/− (KO) and ICSBP+/+ (WT) mice. The frequency of IL-12– producing cells (mean ± SEM) for three individual mice is shown. (C)  RT-PCR analysis of IL-12p40 RNA expression after 6 h of stimulation  of 107 hematopoietic cells with 100 ng/ml LPS ± 100 U/ml IFN-γ. Data  shown are representative of three independent studies. ns, no sample.
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Figure 2: Impaired IL-12 expression in ICSBP−/− mice. (A) Concentration of IL-12p40 in supernatants obtained from indicated cell populations after 18–24 h of incubation with 200 U/ml IFN-γ in combination with 100 ng/ml LPS (left) or 1:1,000 diluted preparation of CD40L (right). Spleen (spl) and bone marrow (bm) cells were plated at 2 × 106/well; normal peritoneal washout (PC) or thioglycollate-elicited exudate (PEX) cells were used at 6 × 105/well. In a number of experiments, serial dilution analysis was performed. Data indicate the mean ± SEM of IL-12p40 concentration in supernatants from the indicated cell populations as well as from macrophages (M∅ ) enriched by adherence. The results summarize data from two to six experiments with two to three mice in each experiment. (B) ELISpot analysis of the ability of ODN or E. coli DNA to trigger IL-12p40 secretion by cells from spleen (left) or lymph nodes (right) of ICSBP−/− (KO) and ICSBP+/+ (WT) mice. The frequency of IL-12– producing cells (mean ± SEM) for three individual mice is shown. (C) RT-PCR analysis of IL-12p40 RNA expression after 6 h of stimulation of 107 hematopoietic cells with 100 ng/ml LPS ± 100 U/ml IFN-γ. Data shown are representative of three independent studies. ns, no sample.
Mentions: Although transcripts of IL-12p35 were comparable in ICSBP−/− and ICSBP+/+ mice, transcripts for IL-12p40 were markedly lower in ICSBP-deficient mice (Fig. 1 B; reference 21). IL-12p35 expression is constitutive in many cell types, whereas IL-12p40 is produced after activation of monocytes, macrophages, neutrophils, dendritic cells, and B cells by various stimuli (for review see reference 41). In the first set of experiments, we examined the ability of cells from spleen, peritoneum, or bone marrow, either unseparated or enriched by adherence for macrophages, to secrete IL-12. The cultures were treated with IFN-γ and either LPS or CD40L (Fig. 2 A). Cells from wild-type mice produced consistently higher levels of IL-12p40 than did cells from ICSBP-deficient mice, with the responses from the latter often being at or below the limits of detection.

Bottom Line: In sera of normal mice, immunoglobulin (Ig)G2a antibodies were dominant over IgG1 antibodies, a pattern indicative of a T helper cell type 1 (Th1)-driven response.Infected ICSBP-deficient mice developed fulminant, disseminated leishmaniasis as a result of failure to mount a Th1-mediated curative response, although T cells remained capable of secreting IFN-gamma and macrophages of producing nitric oxide.This indicates that ICSBP is a deciding factor in Th responses governing humoral and cellular immunity through its role in regulating IL-12 expression.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunopathology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-0760, USA. ngiese@atlas.niaid.nih.gov

ABSTRACT
Mice with a mutation of the gene encoding interferon consensus sequence-binding protein (ICSBP) develop a chronic myelogenous leukemia-like syndrome and mount impaired responses to certain viral and bacterial infections. To gain a mechanistic understanding of the contributions of ICSBP to humoral and cellular immunity, we characterized the responses of control and ICSBP-/- mice to infection with influenza A (flu) and Leishmania major (L. major). Mice of both genotypes survived infections with flu, but differed markedly in the isotype distribution of antiflu antibodies. In sera of normal mice, immunoglobulin (Ig)G2a antibodies were dominant over IgG1 antibodies, a pattern indicative of a T helper cell type 1 (Th1)-driven response. In sera of ICSBP-/- mice, however, IgG1 antibodies dominated over IgG2a antibodies, a pattern indicative of a Th2-driven response. The dominance of IgG1 and IgE over IgG2a was detected in the sera of uninfected mice as well. A seeming Th2 bias of ICSBP-deficient mice was also uncovered in their inability to control infection with L. major, where resistance is known to be dependent on IL-12 and IFN-gamma as components of a Th1 response. Infected ICSBP-deficient mice developed fulminant, disseminated leishmaniasis as a result of failure to mount a Th1-mediated curative response, although T cells remained capable of secreting IFN-gamma and macrophages of producing nitric oxide. Compromised Th1 differentiation in ICSBP-/- mice could not be attributed to hyporesponsiveness of CD4(+) T cells to interleukin (IL)-12; however, the ability of uninfected and infected ICSBP-deficient mice to produce IL-12 was markedly impaired. This indicates that ICSBP is a deciding factor in Th responses governing humoral and cellular immunity through its role in regulating IL-12 expression.

Show MeSH
Related in: MedlinePlus