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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.

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.

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Cellular filiation in the Ubow mouse. (A) Schematic describing that when Ubow cells express Cre, they simultaneously lose dTomato and acquire CFP or YFP expression. Applied to a precursor/product relationship and in combination with the proper Cre reporter strain, this system unravels the transitional stage between progenitors and their differentiated cells. (B) Non-chimeric Ubow+/−-CreERT2 mice were injected with a single dose of tamoxifen. Mice were bled every day and the relative proportion of CFP-, YFP-, and dTomato-expressing B220+ B cells were assessed by flow cytometry (same results were observed in CD3+ T cells). Data are representative of 3 different mice from 3 different experiments. (C) Schematic showing the expected phenotypes and colors of MRCs (RANK-Lhi CD21neg) and immature (RANK-Lmed CD21med) and mature FDCs (RANK-Lneg CD21hi) in CD21Cre Ubow mice according to the model in which MRCs give rise to FDCs.
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fig4: Cellular filiation in the Ubow mouse. (A) Schematic describing that when Ubow cells express Cre, they simultaneously lose dTomato and acquire CFP or YFP expression. Applied to a precursor/product relationship and in combination with the proper Cre reporter strain, this system unravels the transitional stage between progenitors and their differentiated cells. (B) Non-chimeric Ubow+/−-CreERT2 mice were injected with a single dose of tamoxifen. Mice were bled every day and the relative proportion of CFP-, YFP-, and dTomato-expressing B220+ B cells were assessed by flow cytometry (same results were observed in CD3+ T cells). Data are representative of 3 different mice from 3 different experiments. (C) Schematic showing the expected phenotypes and colors of MRCs (RANK-Lhi CD21neg) and immature (RANK-Lmed CD21med) and mature FDCs (RANK-Lneg CD21hi) in CD21Cre Ubow mice according to the model in which MRCs give rise to FDCs.

Mentions: To address the affiliation of MRCs and FDCs, we used another feature of the Ubow mouse. In the Ubow mouse, all cells express dTomato and upon Cre action, cells undergo a single recombination event that triggers YFP or CFP expression. Although Cre genetically switches off the production of dTomato in any Ubow-expressing cell, it does not act on the pool of cytoplasmic dTomato protein that should slowly decrease and eventually wash out (Fig. 4 A). In summary, any Ubow cell that expresses Cre should transiently coexpress dTomato and YFP (or CFP) before acquiring its final color (YFP or CFP). As a proof of principle, we treated Ubow+/− CreERT2 mice with a single dose of tamoxifen and followed the colors of their blood lymphocytes over time (Fig. 4 B). At day 0, all lymphocytes exclusively expressed dTomato. 1 d later, dTomato+ lymphocytes started to express CFP or YFP. During the subsequent days, CFP and YFP expression progressively increased in dTomato-expressing cells. The first single, bright YFP+ (or CFP+) cells negative for dTomato expression appeared at day 6 while the remaining transitional cells gradually lost dTomato in an asynchronous manner, probably reflecting a gradual tamoxifen-induced recombination and intrinsic differences in fluorescent protein degradation (Fig. S3). Altogether, these results indicate that, upon Cre activation, any Ubow cell goes through a transitional step characterized by the coexpression of dTomato and CFP or YFP.


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)

Cellular filiation in the Ubow mouse. (A) Schematic describing that when Ubow cells express Cre, they simultaneously lose dTomato and acquire CFP or YFP expression. Applied to a precursor/product relationship and in combination with the proper Cre reporter strain, this system unravels the transitional stage between progenitors and their differentiated cells. (B) Non-chimeric Ubow+/−-CreERT2 mice were injected with a single dose of tamoxifen. Mice were bled every day and the relative proportion of CFP-, YFP-, and dTomato-expressing B220+ B cells were assessed by flow cytometry (same results were observed in CD3+ T cells). Data are representative of 3 different mice from 3 different experiments. (C) Schematic showing the expected phenotypes and colors of MRCs (RANK-Lhi CD21neg) and immature (RANK-Lmed CD21med) and mature FDCs (RANK-Lneg CD21hi) in CD21Cre Ubow mice according to the model in which MRCs give rise to FDCs.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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

fig4: Cellular filiation in the Ubow mouse. (A) Schematic describing that when Ubow cells express Cre, they simultaneously lose dTomato and acquire CFP or YFP expression. Applied to a precursor/product relationship and in combination with the proper Cre reporter strain, this system unravels the transitional stage between progenitors and their differentiated cells. (B) Non-chimeric Ubow+/−-CreERT2 mice were injected with a single dose of tamoxifen. Mice were bled every day and the relative proportion of CFP-, YFP-, and dTomato-expressing B220+ B cells were assessed by flow cytometry (same results were observed in CD3+ T cells). Data are representative of 3 different mice from 3 different experiments. (C) Schematic showing the expected phenotypes and colors of MRCs (RANK-Lhi CD21neg) and immature (RANK-Lmed CD21med) and mature FDCs (RANK-Lneg CD21hi) in CD21Cre Ubow mice according to the model in which MRCs give rise to FDCs.
Mentions: To address the affiliation of MRCs and FDCs, we used another feature of the Ubow mouse. In the Ubow mouse, all cells express dTomato and upon Cre action, cells undergo a single recombination event that triggers YFP or CFP expression. Although Cre genetically switches off the production of dTomato in any Ubow-expressing cell, it does not act on the pool of cytoplasmic dTomato protein that should slowly decrease and eventually wash out (Fig. 4 A). In summary, any Ubow cell that expresses Cre should transiently coexpress dTomato and YFP (or CFP) before acquiring its final color (YFP or CFP). As a proof of principle, we treated Ubow+/− CreERT2 mice with a single dose of tamoxifen and followed the colors of their blood lymphocytes over time (Fig. 4 B). At day 0, all lymphocytes exclusively expressed dTomato. 1 d later, dTomato+ lymphocytes started to express CFP or YFP. During the subsequent days, CFP and YFP expression progressively increased in dTomato-expressing cells. The first single, bright YFP+ (or CFP+) cells negative for dTomato expression appeared at day 6 while the remaining transitional cells gradually lost dTomato in an asynchronous manner, probably reflecting a gradual tamoxifen-induced recombination and intrinsic differences in fluorescent protein degradation (Fig. S3). Altogether, these results indicate that, upon Cre activation, any Ubow cell goes through a transitional step characterized by the coexpression of dTomato and CFP or YFP.

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