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Identification of a committed T cell precursor population in adult human peripheral blood.

Bruno L, Res P, Dessing M, Cella M, Spits H - J. Exp. Med. (1997)

Bottom Line: At a molecular level, they express the pre-T cell receptor alpha (pT alpha) gene, CD3-gamma, CD-delta and CD-epsilon, and RAG-1 recombination enzyme and have initiated rearrangements in the T cell receptor (TCR)-beta locus (D-J).Moreover, low levels of CD3 epsilon protein, but not of TCR-beta chain, can be detected in their cytoplasm.Our results suggest that CD4+CD3- cells identified in peripheral blood are different from CD3-CD4+CD8- thymocytes and may contain precursors of an extrathymic T cell differentiation pathway.

View Article: PubMed Central - PubMed

Affiliation: Basel Institute for Immunology, Switzerland.

ABSTRACT
Here, we report data concerning the discovery in adult human peripheral blood of a precursor cell population able to differentiate into CD4+CD3+ alpha beta + mature T cells. These cells, which represent 0.1-0.5% of total peripheral blood mononuclear cells (PBMC), express substantial levels of CD4, but lack CD3 surface expression. At a molecular level, they express the pre-T cell receptor alpha (pT alpha) gene, CD3-gamma, CD-delta and CD-epsilon, and RAG-1 recombination enzyme and have initiated rearrangements in the T cell receptor (TCR)-beta locus (D-J). Moreover, low levels of CD3 epsilon protein, but not of TCR-beta chain, can be detected in their cytoplasm. Our results suggest that CD4+CD3- cells identified in peripheral blood are different from CD3-CD4+CD8- thymocytes and may contain precursors of an extrathymic T cell differentiation pathway.

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The capacities of CD4+CD3−CD14− PBMC and CD4+  CD3−CD8−thymocytes of developing into T cells in an FTOC. PBMC  and thymocytes were cultured for 3 wk with mouse RAG-1 −/− thymic  lobes as indicated in Materials and Methods and analyzed with PE, FITC,  and tricolor-labeled mAbs against CD45 (to discriminate between mouse  and human cells) CD3, CD4, and CD8.
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Figure 7: The capacities of CD4+CD3−CD14− PBMC and CD4+ CD3−CD8−thymocytes of developing into T cells in an FTOC. PBMC and thymocytes were cultured for 3 wk with mouse RAG-1 −/− thymic lobes as indicated in Materials and Methods and analyzed with PE, FITC, and tricolor-labeled mAbs against CD45 (to discriminate between mouse and human cells) CD3, CD4, and CD8.

Mentions: The fact that the CD4+CD3− CD14− PB cell population was found to express CD3-γ, CD3-δ, and CD3-ε, pTα, and RAG-1 transcripts, and has partially rearranged the TCR-β locus strongly suggested that committed T cell precursors were present. Moreover, the phenotype of these cells resembles that of the CD3− CD4+CD8− ISP thymocytes. These latter cells differentiate into DP and CD4+TCR-αβ+ cells in mouse FTOC. To analyze the T cell developmental potential, CD4+CD3− CD14− peripheral blood cells were introduced into FTOC. After 3 wk of incubation only few human cells were recovered from the FTOC with CD4+CD3−CD14− peripheral blood cells. To determine accurately the phenotype of these cells, we stained with anti-CD45, anti-CD4, and anti-CD3 or anti-CD8. Fig. 7 shows that almost all human cells expressed high levels of CD3 and CD4. The CD4+ cells completely lack CD8. This pattern of differentiation is distinct from what we observe with CD3−CD4+CD8− ISP thymocytes, which give rise to a majority of DP and SP cells (Fig. 7).


Identification of a committed T cell precursor population in adult human peripheral blood.

Bruno L, Res P, Dessing M, Cella M, Spits H - J. Exp. Med. (1997)

The capacities of CD4+CD3−CD14− PBMC and CD4+  CD3−CD8−thymocytes of developing into T cells in an FTOC. PBMC  and thymocytes were cultured for 3 wk with mouse RAG-1 −/− thymic  lobes as indicated in Materials and Methods and analyzed with PE, FITC,  and tricolor-labeled mAbs against CD45 (to discriminate between mouse  and human cells) CD3, CD4, and CD8.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196171&req=5

Figure 7: The capacities of CD4+CD3−CD14− PBMC and CD4+ CD3−CD8−thymocytes of developing into T cells in an FTOC. PBMC and thymocytes were cultured for 3 wk with mouse RAG-1 −/− thymic lobes as indicated in Materials and Methods and analyzed with PE, FITC, and tricolor-labeled mAbs against CD45 (to discriminate between mouse and human cells) CD3, CD4, and CD8.
Mentions: The fact that the CD4+CD3− CD14− PB cell population was found to express CD3-γ, CD3-δ, and CD3-ε, pTα, and RAG-1 transcripts, and has partially rearranged the TCR-β locus strongly suggested that committed T cell precursors were present. Moreover, the phenotype of these cells resembles that of the CD3− CD4+CD8− ISP thymocytes. These latter cells differentiate into DP and CD4+TCR-αβ+ cells in mouse FTOC. To analyze the T cell developmental potential, CD4+CD3− CD14− peripheral blood cells were introduced into FTOC. After 3 wk of incubation only few human cells were recovered from the FTOC with CD4+CD3−CD14− peripheral blood cells. To determine accurately the phenotype of these cells, we stained with anti-CD45, anti-CD4, and anti-CD3 or anti-CD8. Fig. 7 shows that almost all human cells expressed high levels of CD3 and CD4. The CD4+ cells completely lack CD8. This pattern of differentiation is distinct from what we observe with CD3−CD4+CD8− ISP thymocytes, which give rise to a majority of DP and SP cells (Fig. 7).

Bottom Line: At a molecular level, they express the pre-T cell receptor alpha (pT alpha) gene, CD3-gamma, CD-delta and CD-epsilon, and RAG-1 recombination enzyme and have initiated rearrangements in the T cell receptor (TCR)-beta locus (D-J).Moreover, low levels of CD3 epsilon protein, but not of TCR-beta chain, can be detected in their cytoplasm.Our results suggest that CD4+CD3- cells identified in peripheral blood are different from CD3-CD4+CD8- thymocytes and may contain precursors of an extrathymic T cell differentiation pathway.

View Article: PubMed Central - PubMed

Affiliation: Basel Institute for Immunology, Switzerland.

ABSTRACT
Here, we report data concerning the discovery in adult human peripheral blood of a precursor cell population able to differentiate into CD4+CD3+ alpha beta + mature T cells. These cells, which represent 0.1-0.5% of total peripheral blood mononuclear cells (PBMC), express substantial levels of CD4, but lack CD3 surface expression. At a molecular level, they express the pre-T cell receptor alpha (pT alpha) gene, CD3-gamma, CD-delta and CD-epsilon, and RAG-1 recombination enzyme and have initiated rearrangements in the T cell receptor (TCR)-beta locus (D-J). Moreover, low levels of CD3 epsilon protein, but not of TCR-beta chain, can be detected in their cytoplasm. Our results suggest that CD4+CD3- cells identified in peripheral blood are different from CD3-CD4+CD8- thymocytes and may contain precursors of an extrathymic T cell differentiation pathway.

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