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CollagenVI-Cre mice: A new tool to target stromal cells in secondary lymphoid organs

View Article: PubMed Central - PubMed

ABSTRACT

Stromal cells in secondary lymphoid organs (SLOs) are non-hematopoietic cells involved in the regulation of adaptive immune responses. Three major stromal populations have been identified in adult SLOs: fibroblastic reticular cells (FRCs), follicular dendritic cells (FDCs) and marginal reticular cells (MRCs). The properties of these individual populations are not clearly defined, mainly due to the lack of appropriate genetic tools, especially for MRCs. Here, we analyzed stromal cell targeting in SLOs from a transgenic mouse strain that expresses Cre recombinase under the CollagenVI promoter, using lineage tracing approaches. We show that these mice target specifically MRCs and FDCs, but not FRCs in Peyer’s patches and isolated lymphoid follicles in the intestine. In contrast, stromal cells in lymph nodes and the spleen do not express the transgene, which renders ColVI-cre mice ideal for the specific targeting of stromal cells in the gut-associated lymphoid tissue (GALT). This funding further supports the hypothesis of organ-specific stromal precursors in SLOs. Interestingly, in all tissues analyzed, there was also high specificity for perivascular cells, which have been proposed to act as FDC precursors. Taken together, ColVI-Cre mice are a useful new tool for the dissection of MRC- and FDC-specific functions and plasticity in the GALT.

No MeSH data available.


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ColVI-Cre+ cells are present in PPs and co-localize with MRC- and FDC- specific markers.(A) Representative image of a PP from ColVI–Cre, R26mT/mG mice stained for T cells (CD3 in red), B cells (B220 in blue) and Cre-mediated GFP expression. (B–D) Co-localization of Cre-mediated GFP expression with stromal cell markers. MRCs are marked by MAdCAM-1 and TRANCE (B), FDCs by CD35 and CXCL13 (C) and FRCs by ER-TR7 and CCL21 (D). Inserts display higher magnification of selected areas. Scale bars, 75 μm. Several PPs from 5 different transgenic mice were analyzed.
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f1: ColVI-Cre+ cells are present in PPs and co-localize with MRC- and FDC- specific markers.(A) Representative image of a PP from ColVI–Cre, R26mT/mG mice stained for T cells (CD3 in red), B cells (B220 in blue) and Cre-mediated GFP expression. (B–D) Co-localization of Cre-mediated GFP expression with stromal cell markers. MRCs are marked by MAdCAM-1 and TRANCE (B), FDCs by CD35 and CXCL13 (C) and FRCs by ER-TR7 and CCL21 (D). Inserts display higher magnification of selected areas. Scale bars, 75 μm. Several PPs from 5 different transgenic mice were analyzed.

Mentions: Confocal laser-scanning microscopy of PPs from ColVI-Cre × Rosa26mT/mG revealed expression of the ColVI-Cre transgene mainly in stromal cells within the B-cell area and only in a few cells of the T-cell area (Fig. 1A). To better characterize the pattern of the transgene expression, PP sections were further stained with a combination of markers, so as to identify FDC, MRC, and FRC-specific networks. GFP positive MRCs were identified as TRANCE+ and MAdCAM-1+ cells located under the epithelial dome (Fig. 1B). FDCs, detected near the muscularis layer as CD35+ and CXCL13+ cells, also exhibited high GFP expression (Fig. 1C). In contrast, FRCs, identified as cells expressing ERTR7 and CCL21 in the T cell area, were GFP negative (Fig. 1D).


CollagenVI-Cre mice: A new tool to target stromal cells in secondary lymphoid organs
ColVI-Cre+ cells are present in PPs and co-localize with MRC- and FDC- specific markers.(A) Representative image of a PP from ColVI–Cre, R26mT/mG mice stained for T cells (CD3 in red), B cells (B220 in blue) and Cre-mediated GFP expression. (B–D) Co-localization of Cre-mediated GFP expression with stromal cell markers. MRCs are marked by MAdCAM-1 and TRANCE (B), FDCs by CD35 and CXCL13 (C) and FRCs by ER-TR7 and CCL21 (D). Inserts display higher magnification of selected areas. Scale bars, 75 μm. Several PPs from 5 different transgenic mice were analyzed.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5015111&req=5

f1: ColVI-Cre+ cells are present in PPs and co-localize with MRC- and FDC- specific markers.(A) Representative image of a PP from ColVI–Cre, R26mT/mG mice stained for T cells (CD3 in red), B cells (B220 in blue) and Cre-mediated GFP expression. (B–D) Co-localization of Cre-mediated GFP expression with stromal cell markers. MRCs are marked by MAdCAM-1 and TRANCE (B), FDCs by CD35 and CXCL13 (C) and FRCs by ER-TR7 and CCL21 (D). Inserts display higher magnification of selected areas. Scale bars, 75 μm. Several PPs from 5 different transgenic mice were analyzed.
Mentions: Confocal laser-scanning microscopy of PPs from ColVI-Cre × Rosa26mT/mG revealed expression of the ColVI-Cre transgene mainly in stromal cells within the B-cell area and only in a few cells of the T-cell area (Fig. 1A). To better characterize the pattern of the transgene expression, PP sections were further stained with a combination of markers, so as to identify FDC, MRC, and FRC-specific networks. GFP positive MRCs were identified as TRANCE+ and MAdCAM-1+ cells located under the epithelial dome (Fig. 1B). FDCs, detected near the muscularis layer as CD35+ and CXCL13+ cells, also exhibited high GFP expression (Fig. 1C). In contrast, FRCs, identified as cells expressing ERTR7 and CCL21 in the T cell area, were GFP negative (Fig. 1D).

View Article: PubMed Central - PubMed

ABSTRACT

Stromal cells in secondary lymphoid organs (SLOs) are non-hematopoietic cells involved in the regulation of adaptive immune responses. Three major stromal populations have been identified in adult SLOs: fibroblastic reticular cells (FRCs), follicular dendritic cells (FDCs) and marginal reticular cells (MRCs). The properties of these individual populations are not clearly defined, mainly due to the lack of appropriate genetic tools, especially for MRCs. Here, we analyzed stromal cell targeting in SLOs from a transgenic mouse strain that expresses Cre recombinase under the CollagenVI promoter, using lineage tracing approaches. We show that these mice target specifically MRCs and FDCs, but not FRCs in Peyer’s patches and isolated lymphoid follicles in the intestine. In contrast, stromal cells in lymph nodes and the spleen do not express the transgene, which renders ColVI-cre mice ideal for the specific targeting of stromal cells in the gut-associated lymphoid tissue (GALT). This funding further supports the hypothesis of organ-specific stromal precursors in SLOs. Interestingly, in all tissues analyzed, there was also high specificity for perivascular cells, which have been proposed to act as FDC precursors. Taken together, ColVI-Cre mice are a useful new tool for the dissection of MRC- and FDC-specific functions and plasticity in the GALT.

No MeSH data available.


Related in: MedlinePlus