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NF-kappa B RelA-deficient lymphocytes: normal development of T cells and B cells, impaired production of IgA and IgG1 and reduced proliferative responses.

Doi TS, Takahashi T, Taguchi O, Azuma T, Obata Y - J. Exp. Med. (1997)

Bottom Line: However, the secretion of IgG1 and IgA was reduced in RelA-deficient B cells.Furthermore, both T and B cells lacking RelA showed marked reduction in proliferative responses to stimulation with Con A, anti-CD3, anti-CD3 + anti-CD28, LPS, anti-IgM, and PMA + calcium ionophore.The results indicate that RelA plays a critical role in production of specific Ig isotypes and also in signal transduction pathways for lymphocyte proliferation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.

ABSTRACT
To investigate the function of NF-kappa B RelA (p65), we generated mice deficient in this NF-kappa B family member by homologous recombination. Mice lacking RelA showed liver degeneration and died around embryonic day 14.5. To elucidate the role of RelA in lymphocyte development and function, we transplanted fetal liver cells of 13.5-day embryos from heterozygote matings into irradiated SCID mice. Within 4 weeks, both T and B cells had developed in the SCID mice receiving relA-/- fetal liver transplants, similar to the relA+/+ and +/- cases. T cells were found to mature to Thy-1+/TCR alpha beta +/CD3+/CD4+ or CD8+, while B cells had the ability to differentiate to IgM+/B220+ and to secrete immunoglobulins. However, the secretion of IgG1 and IgA was reduced in RelA-deficient B cells. Furthermore, both T and B cells lacking RelA showed marked reduction in proliferative responses to stimulation with Con A, anti-CD3, anti-CD3 + anti-CD28, LPS, anti-IgM, and PMA + calcium ionophore. The results indicate that RelA plays a critical role in production of specific Ig isotypes and also in signal transduction pathways for lymphocyte proliferation.

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DNA and RNA  blot analyses of the spleen cells of  the transplanted SCID mice. (A)  DNA blot analysis. DNA digested by PstI was analyzed with  a 5′ franking probe (see Fig. 1);  the wild-type relA allele yielding  a 4.1-kb fragment, and the mutant allele a 2.2-kb fragment. (B)  RNA blot analysis. Note the lack  of relA transcripts in the spleen  cells of mice transplanted with  relA−/− fetal liver cells. The  relA−/−, +/+, and +/−  spleen cells express similar  amounts of c-rel, relB, and p50  transcripts.
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Figure 4: DNA and RNA blot analyses of the spleen cells of the transplanted SCID mice. (A) DNA blot analysis. DNA digested by PstI was analyzed with a 5′ franking probe (see Fig. 1); the wild-type relA allele yielding a 4.1-kb fragment, and the mutant allele a 2.2-kb fragment. (B) RNA blot analysis. Note the lack of relA transcripts in the spleen cells of mice transplanted with relA−/− fetal liver cells. The relA−/−, +/+, and +/− spleen cells express similar amounts of c-rel, relB, and p50 transcripts.

Mentions: During normal embryonic development, hematopoietic stem cells emerge in the fetal liver on ED9.5 (36). To test whether the RelA-deficient hematopoietic stem cells can develop in the fetal liver and also whether they can differentiate to mature lymphocytes, fetal liver cells of ED13.5 embryos were transplanted into irradiated SCID mice. Transplantation of fetal liver cells not only from relA+/+ and +/− but also from relA−/− embryos greatly increased the number of cells in the thymus, spleen and lymph nodes of SCID mice. The numbers of cells in spleen and lymph nodes of the mice transplanted with relA−/− fetal liver cells were similar to those receiving either relA+/+ or +/− fetal liver cells. The thymus from mice transplanted with relA−/− fetal liver cells contained fewer cells than those with wild-type or heterozygous fetal liver cells. The origin of the lymphocytes in transplanted mice was determined by testing the expression of H-2Kb antigen. The fetal liver cells were from crosses between 129 and B6, both of which express H-2KbDb, while the recipient SCID themselves express H-2KdDd. As shown in Fig. 3, >95% of lymphocytes of mice transplanted with fetal liver cells expressed H-2Kb, indicating that they were definitely of fetal liver origin. The donor origin of the lymphocytes in transplanted mice with fetal liver cells was further confirmed by the presence of disrupted relA genes and the absence of relA transcripts (Fig. 4). Thus, these results indicated that RelA-deficient hematopoietic stem cells can indeed develop in the fetal liver and also proliferate in SCID mice.


NF-kappa B RelA-deficient lymphocytes: normal development of T cells and B cells, impaired production of IgA and IgG1 and reduced proliferative responses.

Doi TS, Takahashi T, Taguchi O, Azuma T, Obata Y - J. Exp. Med. (1997)

DNA and RNA  blot analyses of the spleen cells of  the transplanted SCID mice. (A)  DNA blot analysis. DNA digested by PstI was analyzed with  a 5′ franking probe (see Fig. 1);  the wild-type relA allele yielding  a 4.1-kb fragment, and the mutant allele a 2.2-kb fragment. (B)  RNA blot analysis. Note the lack  of relA transcripts in the spleen  cells of mice transplanted with  relA−/− fetal liver cells. The  relA−/−, +/+, and +/−  spleen cells express similar  amounts of c-rel, relB, and p50  transcripts.
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Figure 4: DNA and RNA blot analyses of the spleen cells of the transplanted SCID mice. (A) DNA blot analysis. DNA digested by PstI was analyzed with a 5′ franking probe (see Fig. 1); the wild-type relA allele yielding a 4.1-kb fragment, and the mutant allele a 2.2-kb fragment. (B) RNA blot analysis. Note the lack of relA transcripts in the spleen cells of mice transplanted with relA−/− fetal liver cells. The relA−/−, +/+, and +/− spleen cells express similar amounts of c-rel, relB, and p50 transcripts.
Mentions: During normal embryonic development, hematopoietic stem cells emerge in the fetal liver on ED9.5 (36). To test whether the RelA-deficient hematopoietic stem cells can develop in the fetal liver and also whether they can differentiate to mature lymphocytes, fetal liver cells of ED13.5 embryos were transplanted into irradiated SCID mice. Transplantation of fetal liver cells not only from relA+/+ and +/− but also from relA−/− embryos greatly increased the number of cells in the thymus, spleen and lymph nodes of SCID mice. The numbers of cells in spleen and lymph nodes of the mice transplanted with relA−/− fetal liver cells were similar to those receiving either relA+/+ or +/− fetal liver cells. The thymus from mice transplanted with relA−/− fetal liver cells contained fewer cells than those with wild-type or heterozygous fetal liver cells. The origin of the lymphocytes in transplanted mice was determined by testing the expression of H-2Kb antigen. The fetal liver cells were from crosses between 129 and B6, both of which express H-2KbDb, while the recipient SCID themselves express H-2KdDd. As shown in Fig. 3, >95% of lymphocytes of mice transplanted with fetal liver cells expressed H-2Kb, indicating that they were definitely of fetal liver origin. The donor origin of the lymphocytes in transplanted mice with fetal liver cells was further confirmed by the presence of disrupted relA genes and the absence of relA transcripts (Fig. 4). Thus, these results indicated that RelA-deficient hematopoietic stem cells can indeed develop in the fetal liver and also proliferate in SCID mice.

Bottom Line: However, the secretion of IgG1 and IgA was reduced in RelA-deficient B cells.Furthermore, both T and B cells lacking RelA showed marked reduction in proliferative responses to stimulation with Con A, anti-CD3, anti-CD3 + anti-CD28, LPS, anti-IgM, and PMA + calcium ionophore.The results indicate that RelA plays a critical role in production of specific Ig isotypes and also in signal transduction pathways for lymphocyte proliferation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.

ABSTRACT
To investigate the function of NF-kappa B RelA (p65), we generated mice deficient in this NF-kappa B family member by homologous recombination. Mice lacking RelA showed liver degeneration and died around embryonic day 14.5. To elucidate the role of RelA in lymphocyte development and function, we transplanted fetal liver cells of 13.5-day embryos from heterozygote matings into irradiated SCID mice. Within 4 weeks, both T and B cells had developed in the SCID mice receiving relA-/- fetal liver transplants, similar to the relA+/+ and +/- cases. T cells were found to mature to Thy-1+/TCR alpha beta +/CD3+/CD4+ or CD8+, while B cells had the ability to differentiate to IgM+/B220+ and to secrete immunoglobulins. However, the secretion of IgG1 and IgA was reduced in RelA-deficient B cells. Furthermore, both T and B cells lacking RelA showed marked reduction in proliferative responses to stimulation with Con A, anti-CD3, anti-CD3 + anti-CD28, LPS, anti-IgM, and PMA + calcium ionophore. The results indicate that RelA plays a critical role in production of specific Ig isotypes and also in signal transduction pathways for lymphocyte proliferation.

Show MeSH
Related in: MedlinePlus