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Fate mapping reveals origin and dynamics of lymph node follicular dendritic cells.

Jarjour M, Jorquera A, Mondor I, Wienert S, Narang P, Coles MC, Klauschen F, Bajénoff M - J. Exp. Med. (2014)

Bottom Line: In vitro and ex vivo methods, therefore, allow only limited understanding of the genuine immunobiology of FDCs in their native habitat.Herein, we used various multicolor fate mapping systems to investigate the ontogeny and dynamics of lymph node (LN) FDCs in situ.We further demonstrate that during an immune response, FDCs accumulate in germinal centers and that neither the recruitment of circulating progenitors nor the division of local mature FDCs significantly contributes to this accumulation.

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Affiliation: Centre d'Immunologie de Marseille-Luminy (CIML), Aix-Marseille Université, UM2 Marseille, France Institut National de la Santé et de la Recherche Médicale (INSERM), UMR_S 1104 Marseille, France Centre National de la Recherche Scientifique (CNRS), UMR7280 Marseille, France Aix-Marseille Univ (AMU), F-13284 Marseille, France.

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Tracking the ontogeny of FDCs. (A) Schematic showing that in Wnt-1Cre Ubow+/− and +/+ mice, neural crest–derived head and neck mesenchymal cells stochastically express YFP or CFP at different ratios while the rest of the cells, including all hematopoietic cells, express dTomato (Ghigo et al., 2013). (B) Auricular and popliteal LN sections from nonirradiated Wnt-1Cre Ubow+/− and +/+ mice were stained for CD21/35 expression and analyzed by confocal microscopy. F indicates monocolored foci of FDCs. Insets display high-magnification views of FDCs. (C) All confocal pictures from the auricular/cervical LNs of Wnt-1Cre Ubow+/+ mice were processed to generate Voronoi tessellated pictures (see Materials and methods and Fig. S1) amenable to computational simulation. (D) The mean numbers of the rarest colored CD21/35+ FDCs (i.e., CFP++ FDCs) per CFP++ cluster were then compared with Monte Carlo–simulated pictures in which the same colored FDC populations were randomly distributed within the same region of interest (2 simulations displayed out of 10,000). Horizontal bar = median. Each dot represents one follicle. (E) CD21Cre Ubow+/− and +/+ mice were irradiated and reconstituted with WT BM cells to generate chimeric mice in which B cell follicles would be composed of colored FDCs and nonfluorescent B cells. 8 wk later, their peripheral LNs were sectioned, stained for CD21/35 and IgD expression, and imaged by confocal microscopy. (F) Quantification of CFP++ CD21/35+ FDC clustering index in the follicles of CD21Cre Ubow+/+ chimera. Horizontal bar = median. Each dot represents one follicle. In D and F, a two-tailed Student’s t test was used to determine significance. *, P < 0.05. n.s. = nonsignificant. Bars, 25m. Data are representative of 4 different experiments (2 mice per experiment, at least 4 analyzed LNs per mouse).
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fig1: Tracking the ontogeny of FDCs. (A) Schematic showing that in Wnt-1Cre Ubow+/− and +/+ mice, neural crest–derived head and neck mesenchymal cells stochastically express YFP or CFP at different ratios while the rest of the cells, including all hematopoietic cells, express dTomato (Ghigo et al., 2013). (B) Auricular and popliteal LN sections from nonirradiated Wnt-1Cre Ubow+/− and +/+ mice were stained for CD21/35 expression and analyzed by confocal microscopy. F indicates monocolored foci of FDCs. Insets display high-magnification views of FDCs. (C) All confocal pictures from the auricular/cervical LNs of Wnt-1Cre Ubow+/+ mice were processed to generate Voronoi tessellated pictures (see Materials and methods and Fig. S1) amenable to computational simulation. (D) The mean numbers of the rarest colored CD21/35+ FDCs (i.e., CFP++ FDCs) per CFP++ cluster were then compared with Monte Carlo–simulated pictures in which the same colored FDC populations were randomly distributed within the same region of interest (2 simulations displayed out of 10,000). Horizontal bar = median. Each dot represents one follicle. (E) CD21Cre Ubow+/− and +/+ mice were irradiated and reconstituted with WT BM cells to generate chimeric mice in which B cell follicles would be composed of colored FDCs and nonfluorescent B cells. 8 wk later, their peripheral LNs were sectioned, stained for CD21/35 and IgD expression, and imaged by confocal microscopy. (F) Quantification of CFP++ CD21/35+ FDC clustering index in the follicles of CD21Cre Ubow+/+ chimera. Horizontal bar = median. Each dot represents one follicle. In D and F, a two-tailed Student’s t test was used to determine significance. *, P < 0.05. n.s. = nonsignificant. Bars, 25m. Data are representative of 4 different experiments (2 mice per experiment, at least 4 analyzed LNs per mouse).

Mentions: To investigate the ontogeny of LN FDC networks and unravel the nature of their progenitors, we crossed the Ubow mouse to the Wnt-1Cre transgenic mouse, widely used to map the fate of neural crest cells, a transient, multipotent, migratory cell population that gives rise to the head and neck mesenchymal structures (Jiang et al., 2000; Fig. 1 A). If FDCs arose from local mesenchymal progenitors, we expected the Wnt-1Cre Ubow+/− upper LN (auricular and cervical) FDC networks to express CFP and YFP while the lower LN (inguinal and popliteal) FDC networks should continue to express dTomato in absence of Cre expression. However, if neural crest–derived cells migrate after their mesenchymal differentiation beyond the head- and neck-related organs, CFP and YFP FDCs should also be observed in the lower LNs of these mice. Confocal analysis of Wnt-1Cre Ubow+/− mice upper LNs revealed that all mature CD21/35+ FDCs expressed CFP (30%) or YFP (70%; Fig. 1, A and B, left).


Fate mapping reveals origin and dynamics of lymph node follicular dendritic cells.

Jarjour M, Jorquera A, Mondor I, Wienert S, Narang P, Coles MC, Klauschen F, Bajénoff M - J. Exp. Med. (2014)

Tracking the ontogeny of FDCs. (A) Schematic showing that in Wnt-1Cre Ubow+/− and +/+ mice, neural crest–derived head and neck mesenchymal cells stochastically express YFP or CFP at different ratios while the rest of the cells, including all hematopoietic cells, express dTomato (Ghigo et al., 2013). (B) Auricular and popliteal LN sections from nonirradiated Wnt-1Cre Ubow+/− and +/+ mice were stained for CD21/35 expression and analyzed by confocal microscopy. F indicates monocolored foci of FDCs. Insets display high-magnification views of FDCs. (C) All confocal pictures from the auricular/cervical LNs of Wnt-1Cre Ubow+/+ mice were processed to generate Voronoi tessellated pictures (see Materials and methods and Fig. S1) amenable to computational simulation. (D) The mean numbers of the rarest colored CD21/35+ FDCs (i.e., CFP++ FDCs) per CFP++ cluster were then compared with Monte Carlo–simulated pictures in which the same colored FDC populations were randomly distributed within the same region of interest (2 simulations displayed out of 10,000). Horizontal bar = median. Each dot represents one follicle. (E) CD21Cre Ubow+/− and +/+ mice were irradiated and reconstituted with WT BM cells to generate chimeric mice in which B cell follicles would be composed of colored FDCs and nonfluorescent B cells. 8 wk later, their peripheral LNs were sectioned, stained for CD21/35 and IgD expression, and imaged by confocal microscopy. (F) Quantification of CFP++ CD21/35+ FDC clustering index in the follicles of CD21Cre Ubow+/+ chimera. Horizontal bar = median. Each dot represents one follicle. In D and F, a two-tailed Student’s t test was used to determine significance. *, P < 0.05. n.s. = nonsignificant. Bars, 25m. Data are representative of 4 different experiments (2 mice per experiment, at least 4 analyzed LNs per mouse).
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4042641&req=5

fig1: Tracking the ontogeny of FDCs. (A) Schematic showing that in Wnt-1Cre Ubow+/− and +/+ mice, neural crest–derived head and neck mesenchymal cells stochastically express YFP or CFP at different ratios while the rest of the cells, including all hematopoietic cells, express dTomato (Ghigo et al., 2013). (B) Auricular and popliteal LN sections from nonirradiated Wnt-1Cre Ubow+/− and +/+ mice were stained for CD21/35 expression and analyzed by confocal microscopy. F indicates monocolored foci of FDCs. Insets display high-magnification views of FDCs. (C) All confocal pictures from the auricular/cervical LNs of Wnt-1Cre Ubow+/+ mice were processed to generate Voronoi tessellated pictures (see Materials and methods and Fig. S1) amenable to computational simulation. (D) The mean numbers of the rarest colored CD21/35+ FDCs (i.e., CFP++ FDCs) per CFP++ cluster were then compared with Monte Carlo–simulated pictures in which the same colored FDC populations were randomly distributed within the same region of interest (2 simulations displayed out of 10,000). Horizontal bar = median. Each dot represents one follicle. (E) CD21Cre Ubow+/− and +/+ mice were irradiated and reconstituted with WT BM cells to generate chimeric mice in which B cell follicles would be composed of colored FDCs and nonfluorescent B cells. 8 wk later, their peripheral LNs were sectioned, stained for CD21/35 and IgD expression, and imaged by confocal microscopy. (F) Quantification of CFP++ CD21/35+ FDC clustering index in the follicles of CD21Cre Ubow+/+ chimera. Horizontal bar = median. Each dot represents one follicle. In D and F, a two-tailed Student’s t test was used to determine significance. *, P < 0.05. n.s. = nonsignificant. Bars, 25m. Data are representative of 4 different experiments (2 mice per experiment, at least 4 analyzed LNs per mouse).
Mentions: To investigate the ontogeny of LN FDC networks and unravel the nature of their progenitors, we crossed the Ubow mouse to the Wnt-1Cre transgenic mouse, widely used to map the fate of neural crest cells, a transient, multipotent, migratory cell population that gives rise to the head and neck mesenchymal structures (Jiang et al., 2000; Fig. 1 A). If FDCs arose from local mesenchymal progenitors, we expected the Wnt-1Cre Ubow+/− upper LN (auricular and cervical) FDC networks to express CFP and YFP while the lower LN (inguinal and popliteal) FDC networks should continue to express dTomato in absence of Cre expression. However, if neural crest–derived cells migrate after their mesenchymal differentiation beyond the head- and neck-related organs, CFP and YFP FDCs should also be observed in the lower LNs of these mice. Confocal analysis of Wnt-1Cre Ubow+/− mice upper LNs revealed that all mature CD21/35+ FDCs expressed CFP (30%) or YFP (70%; Fig. 1, A and B, left).

Bottom Line: In vitro and ex vivo methods, therefore, allow only limited understanding of the genuine immunobiology of FDCs in their native habitat.Herein, we used various multicolor fate mapping systems to investigate the ontogeny and dynamics of lymph node (LN) FDCs in situ.We further demonstrate that during an immune response, FDCs accumulate in germinal centers and that neither the recruitment of circulating progenitors nor the division of local mature FDCs significantly contributes to this accumulation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centre d'Immunologie de Marseille-Luminy (CIML), Aix-Marseille Université, UM2 Marseille, France Institut National de la Santé et de la Recherche Médicale (INSERM), UMR_S 1104 Marseille, France Centre National de la Recherche Scientifique (CNRS), UMR7280 Marseille, France Aix-Marseille Univ (AMU), F-13284 Marseille, France.

Show MeSH
Related in: MedlinePlus