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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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Related in: MedlinePlus

The fetal liver origin of lymphocytes in transplanted SCID  mice. By transplantation of the fetal liver from relA+/+, +/−, or −/−  ED 13.5 embryos into irradiated SCID mice, the number of lymphocytes  increased. Average numbers of the cells in the spleen, lymph nodes, and  thymus in groups of three mice receiving transplants are indicated above  the distribution plots. Staining in the presence and absence of mAb to H-2Kb  is indicated by solid and open curves, respectively. Flow cytometric analysis of spleen cells, lymph node cells, and thymocytes showed the expression of H-2Kb, indicating the fetal liver origin.
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Figure 3: The fetal liver origin of lymphocytes in transplanted SCID mice. By transplantation of the fetal liver from relA+/+, +/−, or −/− ED 13.5 embryos into irradiated SCID mice, the number of lymphocytes increased. Average numbers of the cells in the spleen, lymph nodes, and thymus in groups of three mice receiving transplants are indicated above the distribution plots. Staining in the presence and absence of mAb to H-2Kb is indicated by solid and open curves, respectively. Flow cytometric analysis of spleen cells, lymph node cells, and thymocytes showed the expression of H-2Kb, indicating the fetal liver origin.

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)

The fetal liver origin of lymphocytes in transplanted SCID  mice. By transplantation of the fetal liver from relA+/+, +/−, or −/−  ED 13.5 embryos into irradiated SCID mice, the number of lymphocytes  increased. Average numbers of the cells in the spleen, lymph nodes, and  thymus in groups of three mice receiving transplants are indicated above  the distribution plots. Staining in the presence and absence of mAb to H-2Kb  is indicated by solid and open curves, respectively. Flow cytometric analysis of spleen cells, lymph node cells, and thymocytes showed the expression of H-2Kb, indicating the fetal liver origin.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196168&req=5

Figure 3: The fetal liver origin of lymphocytes in transplanted SCID mice. By transplantation of the fetal liver from relA+/+, +/−, or −/− ED 13.5 embryos into irradiated SCID mice, the number of lymphocytes increased. Average numbers of the cells in the spleen, lymph nodes, and thymus in groups of three mice receiving transplants are indicated above the distribution plots. Staining in the presence and absence of mAb to H-2Kb is indicated by solid and open curves, respectively. Flow cytometric analysis of spleen cells, lymph node cells, and thymocytes showed the expression of H-2Kb, indicating the fetal liver origin.
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