Single-cell analysis of thymocyte differentiation: identification of transcription factor interactions and a major stochastic component in $\alpha\beta$-lineage commitment. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue PLoS ONE Année : 2013

Single-cell analysis of thymocyte differentiation: identification of transcription factor interactions and a major stochastic component in $\alpha\beta$-lineage commitment.

Résumé

T cell commitment and $\alpha\beta$/$\gamma\delta$ lineage specification in the thymus involves interactions between many different genes. Characterization of these interactions thus requires a multiparameter analysis of individual thymocytes. We developed two efficient single-cell methods: (i) the quantitative evaluation of the co-expression levels of nine different genes, with a plating efficiency of 99-100% and a detection limit of 2 mRNA molecules/cell; and (ii) single-cell differentiation cultures, in the presence of OP9 cells transfected with the thymus Notch1 ligand DeltaL4. We show that during T cell commitment, Gata3 has a fundamental, dose-dependent role in maintaining Notch1 expression, with thymocytes becoming T-cell-committed when they co-express Notch1, Gata3 and Bc11b. Of the transcription factor expression patterns studied here, only that of Bcl11b was suggestive of a role in Pu1 down-regulation. Individual thymocytes became $\alpha\beta$$\gamma\delta$-lineage-committed at very different stages (from the TN2a stage onwards). However, 20% of TN3 cells are not $\alpha\beta$/$\gamma\delta$ lineage committed and TN4 cells comprise two main subpopulations with different degrees of maturity. The existence of a correlation between differentiation potential and expression of the pre-TCR showed that 83% of $\alpha\beta$-committed cells do not express the pre-TCR and revealed a major stochastic component in $\alpha\beta$-lineage specification.

Domaines

Immunologie
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Dates et versions

hal-01070497 , version 1 (11-12-2018)

Identifiants

Citer

Amine Boudil, Lamia Skhiri, Serge Candéias, Valérie Pasqualetto, Agnès Legrand, et al.. Single-cell analysis of thymocyte differentiation: identification of transcription factor interactions and a major stochastic component in $\alpha\beta$-lineage commitment.. PLoS ONE, 2013, 8 (10), pp.e73098. ⟨10.1371/journal.pone.0073098⟩. ⟨hal-01070497⟩
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