Limits...
Gastric hyperplasia and increased proliferative responses of lymphocytes in mice lacking the COOH-terminal ankyrin domain of NF-kappaB2.

Ishikawa H, Carrasco D, Claudio E, Ryseck RP, Bravo R - J. Exp. Med. (1997)

Bottom Line: However, the physiological relevance of the p100 precursor as an IkappaB molecule has not been understood.Dramatic induction of nuclear kappaB-binding activity composed of p52-containing complexes was found in all tissues examined and also in stimulated lymphocytes.Thus, the p100 precursor is essential for the proper regulation of p52-containing Rel/NF-kappaB complexes in various cell types and its absence cannot be efficiently compensated for by other IkappaB proteins.

View Article: PubMed Central - PubMed

Affiliation: The Department of Oncology, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543-4000, USA.

ABSTRACT
The nfkb2 gene encodes the p100 precursor which produces the p52 protein after proteolytic cleavage of its COOH-terminal domain. Although the p52 product can act as an alternative subunit of NF-kappaB, the p100 precursor is believed to function as an inhibitor of Rel/NF-kappaB activity by cytoplasmic retention of Rel/NF-kappaB complexes, like other members of the IkappaB family. However, the physiological relevance of the p100 precursor as an IkappaB molecule has not been understood. To assess the role of the precursor in vivo, we generated, by gene targeting, mice lacking p100 but still containing a functional p52 protein. Mice with a homozygous deletion of the COOH-terminal ankyrin repeats of NF-kappaB2 (p100(-/-)) had marked gastric hyperplasia, resulting in early postnatal death. p100(-/-) animals also presented histopathological alterations of hematopoietic tissues, enlarged lymph nodes, increased lymphocyte proliferation in response to several stimuli, and enhanced cytokine production in activated T cells. Dramatic induction of nuclear kappaB-binding activity composed of p52-containing complexes was found in all tissues examined and also in stimulated lymphocytes. Thus, the p100 precursor is essential for the proper regulation of p52-containing Rel/NF-kappaB complexes in various cell types and its absence cannot be efficiently compensated for by other IkappaB proteins.

Show MeSH

Related in: MedlinePlus

Accelerated proliferative responses  and cytokine production in activated T cells of  p100−/− mice. (A) T cell proliferation in vitro.  Peripheral T cells isolated from spleen of 10-d-old wild-type (closed boxes) and p100−/− mice  (open boxes) were treated with either anti-CD3,  anti-CD3 plus anti-CD28, or PMA plus PHA,  followed by [3H]thymidine incorporation. Values of [3H]thymidine incorporation are shown  by mean ± S.D. (B) The cytokine production  from stimulated T cells of p100−/− mice is increased. Splenic T cells isolated from 10-d-old  wild-type (closed boxes) and p100−/− (open boxes)  mice were treated with (+) or without (−)  anti-CD3 and anti-CD28 antibodies for 72 h.  The cytokine levels in the supernatants were  determined by ELISA. Levels of IL-2, IL-4,  IL-10, GM-CSF, and TNF-α produced in  p100−/− T cells relative to control T cells are  represented by mean values ± S.D.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2199059&req=5

Figure 8: Accelerated proliferative responses and cytokine production in activated T cells of p100−/− mice. (A) T cell proliferation in vitro. Peripheral T cells isolated from spleen of 10-d-old wild-type (closed boxes) and p100−/− mice (open boxes) were treated with either anti-CD3, anti-CD3 plus anti-CD28, or PMA plus PHA, followed by [3H]thymidine incorporation. Values of [3H]thymidine incorporation are shown by mean ± S.D. (B) The cytokine production from stimulated T cells of p100−/− mice is increased. Splenic T cells isolated from 10-d-old wild-type (closed boxes) and p100−/− (open boxes) mice were treated with (+) or without (−) anti-CD3 and anti-CD28 antibodies for 72 h. The cytokine levels in the supernatants were determined by ELISA. Levels of IL-2, IL-4, IL-10, GM-CSF, and TNF-α produced in p100−/− T cells relative to control T cells are represented by mean values ± S.D.

Mentions: The enlarged lymph nodes in p100−/− mice aged 2-wk or older suggest that the increase in κB-binding activity promotes lymphocyte proliferation in vivo. Therefore, we analyzed the proliferative responses of purified T cells from 10-d-old wild type and homozygous mutant mouse spleen stimulated with several mitogens by [3H]thymidine incorporation. Anti-CD3, anti-CD3 plus anti-CD28, or PMA plus PHA promoted proliferation of p100−/− T cells two- to sixfold more efficiently than control T cells (Fig. 8 A). LPS treatment also increased proliferation of p100−/− B cells fivefold more efficiently than control B cells (data not shown). These data indicate that proliferative responses of p100−/− lymphocytes are also enhanced in vitro.


Gastric hyperplasia and increased proliferative responses of lymphocytes in mice lacking the COOH-terminal ankyrin domain of NF-kappaB2.

Ishikawa H, Carrasco D, Claudio E, Ryseck RP, Bravo R - J. Exp. Med. (1997)

Accelerated proliferative responses  and cytokine production in activated T cells of  p100−/− mice. (A) T cell proliferation in vitro.  Peripheral T cells isolated from spleen of 10-d-old wild-type (closed boxes) and p100−/− mice  (open boxes) were treated with either anti-CD3,  anti-CD3 plus anti-CD28, or PMA plus PHA,  followed by [3H]thymidine incorporation. Values of [3H]thymidine incorporation are shown  by mean ± S.D. (B) The cytokine production  from stimulated T cells of p100−/− mice is increased. Splenic T cells isolated from 10-d-old  wild-type (closed boxes) and p100−/− (open boxes)  mice were treated with (+) or without (−)  anti-CD3 and anti-CD28 antibodies for 72 h.  The cytokine levels in the supernatants were  determined by ELISA. Levels of IL-2, IL-4,  IL-10, GM-CSF, and TNF-α produced in  p100−/− T cells relative to control T cells are  represented by mean values ± S.D.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199059&req=5

Figure 8: Accelerated proliferative responses and cytokine production in activated T cells of p100−/− mice. (A) T cell proliferation in vitro. Peripheral T cells isolated from spleen of 10-d-old wild-type (closed boxes) and p100−/− mice (open boxes) were treated with either anti-CD3, anti-CD3 plus anti-CD28, or PMA plus PHA, followed by [3H]thymidine incorporation. Values of [3H]thymidine incorporation are shown by mean ± S.D. (B) The cytokine production from stimulated T cells of p100−/− mice is increased. Splenic T cells isolated from 10-d-old wild-type (closed boxes) and p100−/− (open boxes) mice were treated with (+) or without (−) anti-CD3 and anti-CD28 antibodies for 72 h. The cytokine levels in the supernatants were determined by ELISA. Levels of IL-2, IL-4, IL-10, GM-CSF, and TNF-α produced in p100−/− T cells relative to control T cells are represented by mean values ± S.D.
Mentions: The enlarged lymph nodes in p100−/− mice aged 2-wk or older suggest that the increase in κB-binding activity promotes lymphocyte proliferation in vivo. Therefore, we analyzed the proliferative responses of purified T cells from 10-d-old wild type and homozygous mutant mouse spleen stimulated with several mitogens by [3H]thymidine incorporation. Anti-CD3, anti-CD3 plus anti-CD28, or PMA plus PHA promoted proliferation of p100−/− T cells two- to sixfold more efficiently than control T cells (Fig. 8 A). LPS treatment also increased proliferation of p100−/− B cells fivefold more efficiently than control B cells (data not shown). These data indicate that proliferative responses of p100−/− lymphocytes are also enhanced in vitro.

Bottom Line: However, the physiological relevance of the p100 precursor as an IkappaB molecule has not been understood.Dramatic induction of nuclear kappaB-binding activity composed of p52-containing complexes was found in all tissues examined and also in stimulated lymphocytes.Thus, the p100 precursor is essential for the proper regulation of p52-containing Rel/NF-kappaB complexes in various cell types and its absence cannot be efficiently compensated for by other IkappaB proteins.

View Article: PubMed Central - PubMed

Affiliation: The Department of Oncology, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543-4000, USA.

ABSTRACT
The nfkb2 gene encodes the p100 precursor which produces the p52 protein after proteolytic cleavage of its COOH-terminal domain. Although the p52 product can act as an alternative subunit of NF-kappaB, the p100 precursor is believed to function as an inhibitor of Rel/NF-kappaB activity by cytoplasmic retention of Rel/NF-kappaB complexes, like other members of the IkappaB family. However, the physiological relevance of the p100 precursor as an IkappaB molecule has not been understood. To assess the role of the precursor in vivo, we generated, by gene targeting, mice lacking p100 but still containing a functional p52 protein. Mice with a homozygous deletion of the COOH-terminal ankyrin repeats of NF-kappaB2 (p100(-/-)) had marked gastric hyperplasia, resulting in early postnatal death. p100(-/-) animals also presented histopathological alterations of hematopoietic tissues, enlarged lymph nodes, increased lymphocyte proliferation in response to several stimuli, and enhanced cytokine production in activated T cells. Dramatic induction of nuclear kappaB-binding activity composed of p52-containing complexes was found in all tissues examined and also in stimulated lymphocytes. Thus, the p100 precursor is essential for the proper regulation of p52-containing Rel/NF-kappaB complexes in various cell types and its absence cannot be efficiently compensated for by other IkappaB proteins.

Show MeSH
Related in: MedlinePlus