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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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Structure of the relA targeting vector. The wild-type mouse  relA allele is shown at the top. The targeting vector is in the middle and  the predicted mutant allele is at the bottom. The area predicted to undergo homologous recombination is indicated by the dotted lines. Exons  are indicated by closed boxes. The probes used for diagnostic DNA blot  analysis are also indicated at the top. Restriction enzyme sites: H, HindIII;  N, NcoI; P, PstI; Sm, SmaI; S, SphI.
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Figure 1: Structure of the relA targeting vector. The wild-type mouse relA allele is shown at the top. The targeting vector is in the middle and the predicted mutant allele is at the bottom. The area predicted to undergo homologous recombination is indicated by the dotted lines. Exons are indicated by closed boxes. The probes used for diagnostic DNA blot analysis are also indicated at the top. Restriction enzyme sites: H, HindIII; N, NcoI; P, PstI; Sm, SmaI; S, SphI.

Mentions: The mouse relA gene was isolated from a C57BL/6 (B6)1 mouse genomic library using a mouse relA cDNA probe (codons 185-277, reference 16). The targeting vector was constructed in pBluescript as shown in Fig. 1. It contained 7 kb of the mouse relA gene including exons 1 to 6, PMC1-neo inserted into the first exon of relA at an NcoI site 3 bp downstream of the translation initiation codon, and the herpes simplex virus-thymidine kinase gene (HSV-tk) flanking at the 3′ end of the relA sequence. We expected that this targeting vector would generate a mutant allele by homologous recombination.


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)

Structure of the relA targeting vector. The wild-type mouse  relA allele is shown at the top. The targeting vector is in the middle and  the predicted mutant allele is at the bottom. The area predicted to undergo homologous recombination is indicated by the dotted lines. Exons  are indicated by closed boxes. The probes used for diagnostic DNA blot  analysis are also indicated at the top. Restriction enzyme sites: H, HindIII;  N, NcoI; P, PstI; Sm, SmaI; S, SphI.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: Structure of the relA targeting vector. The wild-type mouse relA allele is shown at the top. The targeting vector is in the middle and the predicted mutant allele is at the bottom. The area predicted to undergo homologous recombination is indicated by the dotted lines. Exons are indicated by closed boxes. The probes used for diagnostic DNA blot analysis are also indicated at the top. Restriction enzyme sites: H, HindIII; N, NcoI; P, PstI; Sm, SmaI; S, SphI.
Mentions: The mouse relA gene was isolated from a C57BL/6 (B6)1 mouse genomic library using a mouse relA cDNA probe (codons 185-277, reference 16). The targeting vector was constructed in pBluescript as shown in Fig. 1. It contained 7 kb of the mouse relA gene including exons 1 to 6, PMC1-neo inserted into the first exon of relA at an NcoI site 3 bp downstream of the translation initiation codon, and the herpes simplex virus-thymidine kinase gene (HSV-tk) flanking at the 3′ end of the relA sequence. We expected that this targeting vector would generate a mutant allele by homologous recombination.

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