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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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Comparison of the  triple negative thymocyte subsets  from wild type (WT) mice and  CD3-εΔ5/Δ5 mutant mice in the  presence or absence of P14  TCR-β transgene (TCRβ).  Thymocytes were stained with  anti-CD3, -CD4, -CD8, -B220,  -Mac-1, and Gr-1 (all biotinylated and detected with streptavidin tricolor), anti-CD44-PE, and  anti-CD25-FITC. The position  of the window (R1) used to  identify the DN T lineage cells is  shown in the top row for each  type of mouse. In the bottom  row, the DN T lineage cells  were analyzed for the expression  of CD25 and CD44. The percentage of cells found within  each quadrant is indicated.
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Figure 2: Comparison of the triple negative thymocyte subsets from wild type (WT) mice and CD3-εΔ5/Δ5 mutant mice in the presence or absence of P14 TCR-β transgene (TCRβ). Thymocytes were stained with anti-CD3, -CD4, -CD8, -B220, -Mac-1, and Gr-1 (all biotinylated and detected with streptavidin tricolor), anti-CD44-PE, and anti-CD25-FITC. The position of the window (R1) used to identify the DN T lineage cells is shown in the top row for each type of mouse. In the bottom row, the DN T lineage cells were analyzed for the expression of CD25 and CD44. The percentage of cells found within each quadrant is indicated.

Mentions: To determine whether the expression of a TCR-β chain transgene was able to inhibit endogenous Vβ to DβJβ rearrangements in the absence of CD3-ε subunit, CD3-εΔ5/Δ5 mice were crossed with transgenic mice carrying a productively rearranged Vβ8+ TCR-β chain derived from the P14 T cell clone (24, 28). When expressed in a wild-type background, this TCR-β transgene prevented endogenous β-locus gene rearrangements, as judged by the fact that most of the SP thymocytes developing in these mice were Vβ8+ (Fig. 1, compare transgenic [WT TCR-β] and nontransgenic [WT] wild-type panels). As shown in Fig. 1 A, CD3-εΔ5/Δ5 TCR-β mice had thymuses that did not develop past the DN stage and contained absolute cell numbers similar to nontransgenic CD3-εΔ5/Δ5 thymuses. Thus, expression of the P14 TCR-β transgene was unable to restore T cell development in CD3-εΔ5/Δ5 mice. To specify more precisely the effect of the TCR-β transgene on early T cell development, we analyzed the CD44/CD25 profile of wild-type and CD3-εΔ5/Δ5 DN thymocytes that developed in the absence or presence of the P14 TCR-β transgene. To this end, we gated on cells that were negative for CD3, CD4, CD8, B cell– (B220), granulocyte– (Gr-1), and macrophage– (Mac-1) specific markers (2). As shown in Fig. 2, comparison of DN cells from transgenic (WT TCR-β) and nontransgenic (WT) wild-type mice indicated that in the former there was a marked increase in the percentage of CD44−/lowCD25− thymocytes at the expense of their immediate CD44−/lowCD25+ precursors. This finding is in line with previous data showing that TCR-β transgenic mice exhibit CD44−/lowCD25+ cell compartments the size of which are intermediate between those found in nontransgenic and TCR-α/β transgenic mice (19, 20). Such observations have been generally accounted for by the fact that CD44−/lowCD25+ cells equipped with a productively rearranged TCR-β transgene progress on average much more rapidly to the CD44−/lowCD25− stage than their nontransgenic counterparts (29). Interestingly, the DN cells found in the CD3-εΔ5/Δ5 and CD3-εΔ5/Δ5 TCR-β mice were both arrested at the same CD44−/lowCD25+ stage and lacked not only the CD44−/lowCD25− cells proper, but also most of the CD44−/lowCD25low to − intermediates. Thus, it is likely that in the absence of CD3-γδε module, pTα–TCR-βP14 heterodimers were prevented from assembling into functional pre-TCR complexes and unable to rescue the blockade in thymic development observed in CD3-εΔ5/Δ5 mice. Alternatively, the onset of expression of the P14 TCR-β transgene during T cell development may have occurred only after the CD44−/low CD25+ stage and accounted for its failure to rescue T cell development in CD3-εΔ5/Δ5 mice.


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)

Comparison of the  triple negative thymocyte subsets  from wild type (WT) mice and  CD3-εΔ5/Δ5 mutant mice in the  presence or absence of P14  TCR-β transgene (TCRβ).  Thymocytes were stained with  anti-CD3, -CD4, -CD8, -B220,  -Mac-1, and Gr-1 (all biotinylated and detected with streptavidin tricolor), anti-CD44-PE, and  anti-CD25-FITC. The position  of the window (R1) used to  identify the DN T lineage cells is  shown in the top row for each  type of mouse. In the bottom  row, the DN T lineage cells  were analyzed for the expression  of CD25 and CD44. The percentage of cells found within  each quadrant is indicated.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199187&req=5

Figure 2: Comparison of the triple negative thymocyte subsets from wild type (WT) mice and CD3-εΔ5/Δ5 mutant mice in the presence or absence of P14 TCR-β transgene (TCRβ). Thymocytes were stained with anti-CD3, -CD4, -CD8, -B220, -Mac-1, and Gr-1 (all biotinylated and detected with streptavidin tricolor), anti-CD44-PE, and anti-CD25-FITC. The position of the window (R1) used to identify the DN T lineage cells is shown in the top row for each type of mouse. In the bottom row, the DN T lineage cells were analyzed for the expression of CD25 and CD44. The percentage of cells found within each quadrant is indicated.
Mentions: To determine whether the expression of a TCR-β chain transgene was able to inhibit endogenous Vβ to DβJβ rearrangements in the absence of CD3-ε subunit, CD3-εΔ5/Δ5 mice were crossed with transgenic mice carrying a productively rearranged Vβ8+ TCR-β chain derived from the P14 T cell clone (24, 28). When expressed in a wild-type background, this TCR-β transgene prevented endogenous β-locus gene rearrangements, as judged by the fact that most of the SP thymocytes developing in these mice were Vβ8+ (Fig. 1, compare transgenic [WT TCR-β] and nontransgenic [WT] wild-type panels). As shown in Fig. 1 A, CD3-εΔ5/Δ5 TCR-β mice had thymuses that did not develop past the DN stage and contained absolute cell numbers similar to nontransgenic CD3-εΔ5/Δ5 thymuses. Thus, expression of the P14 TCR-β transgene was unable to restore T cell development in CD3-εΔ5/Δ5 mice. To specify more precisely the effect of the TCR-β transgene on early T cell development, we analyzed the CD44/CD25 profile of wild-type and CD3-εΔ5/Δ5 DN thymocytes that developed in the absence or presence of the P14 TCR-β transgene. To this end, we gated on cells that were negative for CD3, CD4, CD8, B cell– (B220), granulocyte– (Gr-1), and macrophage– (Mac-1) specific markers (2). As shown in Fig. 2, comparison of DN cells from transgenic (WT TCR-β) and nontransgenic (WT) wild-type mice indicated that in the former there was a marked increase in the percentage of CD44−/lowCD25− thymocytes at the expense of their immediate CD44−/lowCD25+ precursors. This finding is in line with previous data showing that TCR-β transgenic mice exhibit CD44−/lowCD25+ cell compartments the size of which are intermediate between those found in nontransgenic and TCR-α/β transgenic mice (19, 20). Such observations have been generally accounted for by the fact that CD44−/lowCD25+ cells equipped with a productively rearranged TCR-β transgene progress on average much more rapidly to the CD44−/lowCD25− stage than their nontransgenic counterparts (29). Interestingly, the DN cells found in the CD3-εΔ5/Δ5 and CD3-εΔ5/Δ5 TCR-β mice were both arrested at the same CD44−/lowCD25+ stage and lacked not only the CD44−/lowCD25− cells proper, but also most of the CD44−/lowCD25low to − intermediates. Thus, it is likely that in the absence of CD3-γδε module, pTα–TCR-βP14 heterodimers were prevented from assembling into functional pre-TCR complexes and unable to rescue the blockade in thymic development observed in CD3-εΔ5/Δ5 mice. Alternatively, the onset of expression of the P14 TCR-β transgene during T cell development may have occurred only after the CD44−/low CD25+ stage and accounted for its failure to rescue T cell development in CD3-εΔ5/Δ5 mice.

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.

Show MeSH
Related in: MedlinePlus