Characterization of T cell differentiation in the murine gut.
Gut intraepithelial CD8 T lymphocytes (T-IEL) are distinct from thymus-derived cells and are thought to derive locally from cryptopatch (CP) precursors. The intermediate stages of differentiation between CP and mature T-IEL were not identified, and the local differentiation process was not characterized. We identified and characterized six phenotypically distinct lineage-negative populations in the CP and the gut epithelium: (a) we determined the kinetics of their generation from bone marrow precursors; (b) we quantified CD3-epsilon, recombination activating gene (Rag)-1, and pre-Talpha mRNAs expression at single cell level; (c) we characterized TCR-beta, -gamma, and -alpha locus rearrangements; and (d) we studied the impact of different mutations on the local differentiation. These data allowed us to establish a sequence of T cell precursor differentiation in the gut. We also observed that the gut differentiation varied from that of the thymus by a very low frequency of pre-Talpha chain mRNA expression, a different kinetics of Rag-1 mRNA expression, and a much higher impact of CD3 epsilon/delta and pre-Talpha deficiencies. Finally, only 3% of CP cells were clearly involved in T cell differentiation, suggesting that these structures may have additional physiological roles in the gut.
Lambolez
Florence
F
Institut National de la Santé et de la Recherche Médicale (INSERM) U345, Institut Necker, Rue de Vaugirard, 75730 Paris Cedex 15, France.
Azogui
Orly
O
Joret
Anne-Marie
AM
Garcia
Corinne
C
von Boehmer
Harald
H
Di Santo
James
J
Ezine
Sophie
S
Rocha
Benedita
B
D000818:Animals; D017252:CD3 Complex; D002454:Cell Differentiation; D019070:Cell Lineage; D048429:Cell Size; D004268:DNA-Binding Proteins / Q000235:genetics; D004064:Digestive System / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D015329:Gene Rearrangement, T-Lymphocyte / Q000235:genetics; D016654:Genes, RAG-1 / Q000235:genetics; D019673:Genes, T-Cell Receptor alpha; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism; D013934:Thymectomy; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology / Q000378:metabolism
D017252:CD3 Complex; C115335:CD3E protein, human; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; C499308:Rag2 protein, mouse; D011948:Receptors, Antigen, T-Cell; C064655:V(D)J recombination activating protein 2