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The CD3-gammadeltaepsilon and CD3-zeta/eta modules are each essential for allelic exclusion at the T cell receptor beta locus but are both dispensable for the initiation of V to (D)J recombination at the T cell receptor-beta, -gamma, and -delta loci.

Ardouin L, Ismaili J, Malissen B, Malissen M - J. Exp. Med. (1998)

Bottom Line: The pre-T cell receptor (TCR) associates with CD3-transducing subunits and triggers the selective expansion and maturation of T cell precursors expressing a TCR-beta chain.Furthermore, using mutant mice lacking both the CD3-epsilon and CD3-zeta/eta genes, we established that the CD3 gene products are dispensable for the onset of V to (D)J recombination (V, variable; D, diversity; J, joining) at the TCR-beta, TCR-gamma, and TCR-delta loci.Thus, the CD3 components are differentially involved in the sequential events that make the TCR-beta locus first accessible to, and later insulated from, the action of the V(D)J recombinase.

View Article: PubMed Central - PubMed

Affiliation: Centre d'Immunologie Institut National de la Santé et de la Recherche Médicale-Centre National de la Recherche Scientifique de Marseille-Luminy, Case 906, 13288 Marseille Cedex 9, France.

ABSTRACT
The pre-T cell receptor (TCR) associates with CD3-transducing subunits and triggers the selective expansion and maturation of T cell precursors expressing a TCR-beta chain. Recent experiments in pre-Talpha chain-deficient mice have suggested that the pre-TCR may not be required for signaling allelic exclusion at the TCR-beta locus. Using CD3-epsilon- and CD3-zeta/eta-deficient mice harboring a productively rearranged TCR-beta transgene, we showed that the CD3-gammadeltaepsilon and CD3-zeta/eta modules, and by inference the pre-TCR/CD3 complex, are each essential for the establishment of allelic exclusion at the endogenous TCR-beta locus. Furthermore, using mutant mice lacking both the CD3-epsilon and CD3-zeta/eta genes, we established that the CD3 gene products are dispensable for the onset of V to (D)J recombination (V, variable; D, diversity; J, joining) at the TCR-beta, TCR-gamma, and TCR-delta loci. Thus, the CD3 components are differentially involved in the sequential events that make the TCR-beta locus first accessible to, and later insulated from, the action of the V(D)J recombinase.

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Relative levels of  TCR-γ and TCR-δ gene rearrangements in RAG-1−/−,  CD3-εΔ5/Δ5, CD3-ζ/η−/−,  CD3-εΔ5/Δ5, CD3-ζ/η−/−, and  wild-type thymocytes. DNA extracted from thymocytes of fetuses at day 17 of gestation (wt  E17) and of 4–6-wk-old wild-type (WT) and mutant mice was  amplified with PCR primer pairs  specific for the Vδ1–Jδ2, Vδ4– Jδ1, Vδ5–Jδ1, Vγ5–Jγ1, Vγ4–Jγ1,  and Vγ1–Jγ2 rearrangements.  PCR products were gel fractionated and the corresponding  Southern blots hybridized with  labeled oligonucleotide probes.
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Figure 8: Relative levels of TCR-γ and TCR-δ gene rearrangements in RAG-1−/−, CD3-εΔ5/Δ5, CD3-ζ/η−/−, CD3-εΔ5/Δ5, CD3-ζ/η−/−, and wild-type thymocytes. DNA extracted from thymocytes of fetuses at day 17 of gestation (wt E17) and of 4–6-wk-old wild-type (WT) and mutant mice was amplified with PCR primer pairs specific for the Vδ1–Jδ2, Vδ4– Jδ1, Vδ5–Jδ1, Vγ5–Jγ1, Vγ4–Jγ1, and Vγ1–Jγ2 rearrangements. PCR products were gel fractionated and the corresponding Southern blots hybridized with labeled oligonucleotide probes.

Mentions: Considering that thymocytes that lack both CD3-ε and CD3-ζ/η genes are still capable of reaching the CD44−/low CD25+ DN stage during which Vβ to DβJβ recombination normally happens (Fig. 6 B), we analyzed the status of their TCR-β loci using the DNA-PCR assay previously described in the legend of Fig. 4. As shown in Fig. 7, CD3-εΔ5/Δ5CD3-ζ/η−/− double mutant mice contained Dβ→ Jβ and Vβ→ DβJβ rearrangements, the extent of which was similar to those found in the parental single mutant thymocytes. Finally, we examined the effects of the lack of both CD3-ε and CD3-ζ/η on the rearrangement of TCR-γ and -δ genes using a DNA-PCR approach (27). As shown in Fig. 8, the absence of both CD3-ε and CD3-ζ/η had little effect on the extent and timing of TCR-γ and -δ gene rearrangements.


The CD3-gammadeltaepsilon and CD3-zeta/eta modules are each essential for allelic exclusion at the T cell receptor beta locus but are both dispensable for the initiation of V to (D)J recombination at the T cell receptor-beta, -gamma, and -delta loci.

Ardouin L, Ismaili J, Malissen B, Malissen M - J. Exp. Med. (1998)

Relative levels of  TCR-γ and TCR-δ gene rearrangements in RAG-1−/−,  CD3-εΔ5/Δ5, CD3-ζ/η−/−,  CD3-εΔ5/Δ5, CD3-ζ/η−/−, and  wild-type thymocytes. DNA extracted from thymocytes of fetuses at day 17 of gestation (wt  E17) and of 4–6-wk-old wild-type (WT) and mutant mice was  amplified with PCR primer pairs  specific for the Vδ1–Jδ2, Vδ4– Jδ1, Vδ5–Jδ1, Vγ5–Jγ1, Vγ4–Jγ1,  and Vγ1–Jγ2 rearrangements.  PCR products were gel fractionated and the corresponding  Southern blots hybridized with  labeled oligonucleotide probes.
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Related In: Results  -  Collection

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Figure 8: Relative levels of TCR-γ and TCR-δ gene rearrangements in RAG-1−/−, CD3-εΔ5/Δ5, CD3-ζ/η−/−, CD3-εΔ5/Δ5, CD3-ζ/η−/−, and wild-type thymocytes. DNA extracted from thymocytes of fetuses at day 17 of gestation (wt E17) and of 4–6-wk-old wild-type (WT) and mutant mice was amplified with PCR primer pairs specific for the Vδ1–Jδ2, Vδ4– Jδ1, Vδ5–Jδ1, Vγ5–Jγ1, Vγ4–Jγ1, and Vγ1–Jγ2 rearrangements. PCR products were gel fractionated and the corresponding Southern blots hybridized with labeled oligonucleotide probes.
Mentions: Considering that thymocytes that lack both CD3-ε and CD3-ζ/η genes are still capable of reaching the CD44−/low CD25+ DN stage during which Vβ to DβJβ recombination normally happens (Fig. 6 B), we analyzed the status of their TCR-β loci using the DNA-PCR assay previously described in the legend of Fig. 4. As shown in Fig. 7, CD3-εΔ5/Δ5CD3-ζ/η−/− double mutant mice contained Dβ→ Jβ and Vβ→ DβJβ rearrangements, the extent of which was similar to those found in the parental single mutant thymocytes. Finally, we examined the effects of the lack of both CD3-ε and CD3-ζ/η on the rearrangement of TCR-γ and -δ genes using a DNA-PCR approach (27). As shown in Fig. 8, the absence of both CD3-ε and CD3-ζ/η had little effect on the extent and timing of TCR-γ and -δ gene rearrangements.

Bottom Line: The pre-T cell receptor (TCR) associates with CD3-transducing subunits and triggers the selective expansion and maturation of T cell precursors expressing a TCR-beta chain.Furthermore, using mutant mice lacking both the CD3-epsilon and CD3-zeta/eta genes, we established that the CD3 gene products are dispensable for the onset of V to (D)J recombination (V, variable; D, diversity; J, joining) at the TCR-beta, TCR-gamma, and TCR-delta loci.Thus, the CD3 components are differentially involved in the sequential events that make the TCR-beta locus first accessible to, and later insulated from, the action of the V(D)J recombinase.

View Article: PubMed Central - PubMed

Affiliation: Centre d'Immunologie Institut National de la Santé et de la Recherche Médicale-Centre National de la Recherche Scientifique de Marseille-Luminy, Case 906, 13288 Marseille Cedex 9, France.

ABSTRACT
The pre-T cell receptor (TCR) associates with CD3-transducing subunits and triggers the selective expansion and maturation of T cell precursors expressing a TCR-beta chain. Recent experiments in pre-Talpha chain-deficient mice have suggested that the pre-TCR may not be required for signaling allelic exclusion at the TCR-beta locus. Using CD3-epsilon- and CD3-zeta/eta-deficient mice harboring a productively rearranged TCR-beta transgene, we showed that the CD3-gammadeltaepsilon and CD3-zeta/eta modules, and by inference the pre-TCR/CD3 complex, are each essential for the establishment of allelic exclusion at the endogenous TCR-beta locus. Furthermore, using mutant mice lacking both the CD3-epsilon and CD3-zeta/eta genes, we established that the CD3 gene products are dispensable for the onset of V to (D)J recombination (V, variable; D, diversity; J, joining) at the TCR-beta, TCR-gamma, and TCR-delta loci. Thus, the CD3 components are differentially involved in the sequential events that make the TCR-beta locus first accessible to, and later insulated from, the action of the V(D)J recombinase.

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