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Identical T cell clones are located within the mouse gut epithelium and lamina propia and circulate in the thoracic duct lymph.

Arstila T, Arstila TP, Calbo S, Selz F, Malassis-Seris M, Vassalli P, Kourilsky P, Guy-Grand D - J. Exp. Med. (2000)

Bottom Line: Murine gut intraepithelial (IEL) T cell receptor (TCR)-alpha/beta lymphocytes bearing CD8alpha/13 or CD8alpha/alpha coreceptors have been shown previously to express different oligoclonal TCR beta chain repertoires in the same mouse, in agreement with other evidence indicating that these two populations belong to different ontogenic lineages, with only CD8alpha/beta+ IELs being fully thymus dependent.Furthermore, identical T cell clones were detected among CD8alpha/beta IELs and CD8alpha/beta+ blasts circulating into the thoracic duct (TD) lymph of the same mouse, whereas TD small lymphocytes are polyclonal.These findings must be considered in light of previous observations showing that T blasts, but not small T lymphocytes, circulating in the TD lymph have the capacity of homing into the gut epithelium and lamina propria.

View Article: PubMed Central - PubMed

Affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM), Hôpital Necker-Enfants Malades, Paris, France.

ABSTRACT
Murine gut intraepithelial (IEL) T cell receptor (TCR)-alpha/beta lymphocytes bearing CD8alpha/13 or CD8alpha/alpha coreceptors have been shown previously to express different oligoclonal TCR beta chain repertoires in the same mouse, in agreement with other evidence indicating that these two populations belong to different ontogenic lineages, with only CD8alpha/beta+ IELs being fully thymus dependent. CD8alpha/beta+, but not CD8alpha/alpha+, T lymphocytes are also present in the lamina propria. Here, we show that CD8alpha/beta+ lymphocytes from the lamina propria and the epithelium are both oligoclonal, and that they share the same TCR-beta clonotypes in the same mouse, as is also the case for CD4alpha T cells. Furthermore, identical T cell clones were detected among CD8alpha/beta IELs and CD8alpha/beta+ blasts circulating into the thoracic duct (TD) lymph of the same mouse, whereas TD small lymphocytes are polyclonal. These findings must be considered in light of previous observations showing that T blasts, but not small T lymphocytes, circulating in the TD lymph have the capacity of homing into the gut epithelium and lamina propria. These combined observations have interesting implications for our understanding of the recirculation of gut thymus-dependent lymphocytes and their precursors, and of the events leading up to the selection of their restricted TCR repertoire.

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Gut mucosa of a mouse recipient of the BrdU-labeled TD cells shown in Fig. 6. Gut sections displaying two BrdU-labeled cells localized (right) or as seen by phase–contrast microscopy (left) in the epithelium (top) and in the lamina propria (bottom).
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Figure 7: Gut mucosa of a mouse recipient of the BrdU-labeled TD cells shown in Fig. 6. Gut sections displaying two BrdU-labeled cells localized (right) or as seen by phase–contrast microscopy (left) in the epithelium (top) and in the lamina propria (bottom).

Mentions: To ensure that CD8α/β1CD11c+ cells are indeed blasts capable of homing to the gut wall, as observed in previous experiments 7891011, we performed transfer experiments of CD8α/β1 TD cells obtained under continuous perfusion of BrdU to label cycling cells. Since it is not possible to inject cells coated with mAbs into recipients to explore their physiological tissue homing, we used pools of negatively selected TD cells depleted in CD4+ and B cells (see Materials and Methods). By three-color analysis of the transferred cells, the vast majority (90%) were CD8β1, as was also the case of virtually all BrdU+ cells (2.6% of the selected population; Fig. 6 A); 4.3% of CD8β1 cells were CD11c+ (in agreement with the percentage observed with CD8α/β1CD11c+ selected by sorting for repertoire analyses), and all CD11c+ cells were CD8β1 (Fig. 6 B). About 50% BrdU+ cells were CD11c+, and there was a clear correlation between the level of CD11c expression and the intensity of BrdU labeling of BrdU+ cells, although CD11c+ cells also contain a sizable fraction of BrdU− cells (Fig. 6 D). This last observation was consistent with the results of forward scatter analyses of CD11c+ and CD11c− cells, showing that the CD11c+ population contains larger lymphocytes than the bulk of CD11c− lymphocytes (Fig. 6 C). 24 h after transfer of the negatively selected population just described, BrdU+ cells were detected in all sections of the recipient's gut (where their density ratio versus the spleen was comparable to that described previously [reference 15; Fig. 7]).


Identical T cell clones are located within the mouse gut epithelium and lamina propia and circulate in the thoracic duct lymph.

Arstila T, Arstila TP, Calbo S, Selz F, Malassis-Seris M, Vassalli P, Kourilsky P, Guy-Grand D - J. Exp. Med. (2000)

Gut mucosa of a mouse recipient of the BrdU-labeled TD cells shown in Fig. 6. Gut sections displaying two BrdU-labeled cells localized (right) or as seen by phase–contrast microscopy (left) in the epithelium (top) and in the lamina propria (bottom).
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Related In: Results  -  Collection

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Figure 7: Gut mucosa of a mouse recipient of the BrdU-labeled TD cells shown in Fig. 6. Gut sections displaying two BrdU-labeled cells localized (right) or as seen by phase–contrast microscopy (left) in the epithelium (top) and in the lamina propria (bottom).
Mentions: To ensure that CD8α/β1CD11c+ cells are indeed blasts capable of homing to the gut wall, as observed in previous experiments 7891011, we performed transfer experiments of CD8α/β1 TD cells obtained under continuous perfusion of BrdU to label cycling cells. Since it is not possible to inject cells coated with mAbs into recipients to explore their physiological tissue homing, we used pools of negatively selected TD cells depleted in CD4+ and B cells (see Materials and Methods). By three-color analysis of the transferred cells, the vast majority (90%) were CD8β1, as was also the case of virtually all BrdU+ cells (2.6% of the selected population; Fig. 6 A); 4.3% of CD8β1 cells were CD11c+ (in agreement with the percentage observed with CD8α/β1CD11c+ selected by sorting for repertoire analyses), and all CD11c+ cells were CD8β1 (Fig. 6 B). About 50% BrdU+ cells were CD11c+, and there was a clear correlation between the level of CD11c expression and the intensity of BrdU labeling of BrdU+ cells, although CD11c+ cells also contain a sizable fraction of BrdU− cells (Fig. 6 D). This last observation was consistent with the results of forward scatter analyses of CD11c+ and CD11c− cells, showing that the CD11c+ population contains larger lymphocytes than the bulk of CD11c− lymphocytes (Fig. 6 C). 24 h after transfer of the negatively selected population just described, BrdU+ cells were detected in all sections of the recipient's gut (where their density ratio versus the spleen was comparable to that described previously [reference 15; Fig. 7]).

Bottom Line: Murine gut intraepithelial (IEL) T cell receptor (TCR)-alpha/beta lymphocytes bearing CD8alpha/13 or CD8alpha/alpha coreceptors have been shown previously to express different oligoclonal TCR beta chain repertoires in the same mouse, in agreement with other evidence indicating that these two populations belong to different ontogenic lineages, with only CD8alpha/beta+ IELs being fully thymus dependent.Furthermore, identical T cell clones were detected among CD8alpha/beta IELs and CD8alpha/beta+ blasts circulating into the thoracic duct (TD) lymph of the same mouse, whereas TD small lymphocytes are polyclonal.These findings must be considered in light of previous observations showing that T blasts, but not small T lymphocytes, circulating in the TD lymph have the capacity of homing into the gut epithelium and lamina propria.

View Article: PubMed Central - PubMed

Affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM), Hôpital Necker-Enfants Malades, Paris, France.

ABSTRACT
Murine gut intraepithelial (IEL) T cell receptor (TCR)-alpha/beta lymphocytes bearing CD8alpha/13 or CD8alpha/alpha coreceptors have been shown previously to express different oligoclonal TCR beta chain repertoires in the same mouse, in agreement with other evidence indicating that these two populations belong to different ontogenic lineages, with only CD8alpha/beta+ IELs being fully thymus dependent. CD8alpha/beta+, but not CD8alpha/alpha+, T lymphocytes are also present in the lamina propria. Here, we show that CD8alpha/beta+ lymphocytes from the lamina propria and the epithelium are both oligoclonal, and that they share the same TCR-beta clonotypes in the same mouse, as is also the case for CD4alpha T cells. Furthermore, identical T cell clones were detected among CD8alpha/beta IELs and CD8alpha/beta+ blasts circulating into the thoracic duct (TD) lymph of the same mouse, whereas TD small lymphocytes are polyclonal. These findings must be considered in light of previous observations showing that T blasts, but not small T lymphocytes, circulating in the TD lymph have the capacity of homing into the gut epithelium and lamina propria. These combined observations have interesting implications for our understanding of the recirculation of gut thymus-dependent lymphocytes and their precursors, and of the events leading up to the selection of their restricted TCR repertoire.

Show MeSH