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Haematopoietic stem cells in spleen have distinct differentiative potential for antigen presenting cells.

Tan JK, O'Neill HC - J. Cell. Mol. Med. (2010)

Bottom Line: The ability of HSC in spleen to develop into L-DC was indicated by a strong bias in the subset size of these cells over other splenic APC subsets.This type of evidence supports a model whereby spleen represents an important site for haematopoiesis of this distinct DC subset.The conditions under which haematopoiesis of L-DC occurs in spleen, or the progenitors involved, will require further investigation.

View Article: PubMed Central - PubMed

Affiliation: Research School of Biology, The Australian National University, Canberra, Australia.

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Dendritic cell reconstitution by neonatal spleen HSC. Spleen (SPL) and bone marrow (BM) chimeras were prepared as described in Figure 1. For analysis of long-term (.25 week) reconstitution, spleens were collected, enriched for DC by depletion of T and B cells, and stained with antibodies specific for CD45.1, CD11c, CD11b, CD8 and MHC-II. Prior to flow cytometry, cells were incubated with propidium iodide (PI; 1 mg/ml) for gating of live (PI−) cells. C57BL/6J splenocytes were stained as a CD45.1− control. (A) Individual spleen APC subsets (L-DC, myeloid cells, CD8− cDC and CD8+ cDC) were gated as shown in Figure 1A and assessed for donor-cell (CD45.1+) composition. Numbers in gates indicate per cent positive cells. Labelled chimeras (*) indicate mice showing only partial donor reconstitution with some evidence of host-type cell development. (B) The relative prevalence of donor- to host-type cells was calculated for each spleen APC subset in animals showing partial donor reconstitution as indicated in Figures 1 and 2 (*). The fold-increase of donor- to host-type cells was calculated relative to CD8− cDC assigned a fold-increase of 1.0. Data represent mean 6 SE (n= 4). Cell subsets having significantly higher representation of donor- over host-type cells, compared with CD8− cDC, are indicated (*) (P≤ 0.02; Wilcoxon Rank Sum Test).
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fig02: Dendritic cell reconstitution by neonatal spleen HSC. Spleen (SPL) and bone marrow (BM) chimeras were prepared as described in Figure 1. For analysis of long-term (.25 week) reconstitution, spleens were collected, enriched for DC by depletion of T and B cells, and stained with antibodies specific for CD45.1, CD11c, CD11b, CD8 and MHC-II. Prior to flow cytometry, cells were incubated with propidium iodide (PI; 1 mg/ml) for gating of live (PI−) cells. C57BL/6J splenocytes were stained as a CD45.1− control. (A) Individual spleen APC subsets (L-DC, myeloid cells, CD8− cDC and CD8+ cDC) were gated as shown in Figure 1A and assessed for donor-cell (CD45.1+) composition. Numbers in gates indicate per cent positive cells. Labelled chimeras (*) indicate mice showing only partial donor reconstitution with some evidence of host-type cell development. (B) The relative prevalence of donor- to host-type cells was calculated for each spleen APC subset in animals showing partial donor reconstitution as indicated in Figures 1 and 2 (*). The fold-increase of donor- to host-type cells was calculated relative to CD8− cDC assigned a fold-increase of 1.0. Data represent mean 6 SE (n= 4). Cell subsets having significantly higher representation of donor- over host-type cells, compared with CD8− cDC, are indicated (*) (P≤ 0.02; Wilcoxon Rank Sum Test).

Mentions: The superior reconstitution potential of HSC in neonatal spleen was long-lasting, providing CD45.1+ donor-type haematopoietic cells in all SPL chimeras tested (3 of 3) for more than 25 weeks (Fig. 2A). As with short-term reconstitution analyses shown in Figure 1B, all long-term SPL chimeras gave almost complete donor (CD45.1+) cell reconstitution. This demonstrates that donor-type neonatal spleen cells (CD45.1+) out-compete an equal number of host-type BM cells (CD45.1−) for haematopoietic reconstitution, under the adoptive transfer protocol used here. Complete (.99%) reconstitution of spleen APC subsets by donor-type spleen HSC was observed in two out of three SPL chimeras analysed after 25 weeks. Only one chimera displayed partial donor reconstitution of APC subsets in spleen at 51 weeks, with proportions of subsets varying from 67% to 96%.


Haematopoietic stem cells in spleen have distinct differentiative potential for antigen presenting cells.

Tan JK, O'Neill HC - J. Cell. Mol. Med. (2010)

Dendritic cell reconstitution by neonatal spleen HSC. Spleen (SPL) and bone marrow (BM) chimeras were prepared as described in Figure 1. For analysis of long-term (.25 week) reconstitution, spleens were collected, enriched for DC by depletion of T and B cells, and stained with antibodies specific for CD45.1, CD11c, CD11b, CD8 and MHC-II. Prior to flow cytometry, cells were incubated with propidium iodide (PI; 1 mg/ml) for gating of live (PI−) cells. C57BL/6J splenocytes were stained as a CD45.1− control. (A) Individual spleen APC subsets (L-DC, myeloid cells, CD8− cDC and CD8+ cDC) were gated as shown in Figure 1A and assessed for donor-cell (CD45.1+) composition. Numbers in gates indicate per cent positive cells. Labelled chimeras (*) indicate mice showing only partial donor reconstitution with some evidence of host-type cell development. (B) The relative prevalence of donor- to host-type cells was calculated for each spleen APC subset in animals showing partial donor reconstitution as indicated in Figures 1 and 2 (*). The fold-increase of donor- to host-type cells was calculated relative to CD8− cDC assigned a fold-increase of 1.0. Data represent mean 6 SE (n= 4). Cell subsets having significantly higher representation of donor- over host-type cells, compared with CD8− cDC, are indicated (*) (P≤ 0.02; Wilcoxon Rank Sum Test).
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fig02: Dendritic cell reconstitution by neonatal spleen HSC. Spleen (SPL) and bone marrow (BM) chimeras were prepared as described in Figure 1. For analysis of long-term (.25 week) reconstitution, spleens were collected, enriched for DC by depletion of T and B cells, and stained with antibodies specific for CD45.1, CD11c, CD11b, CD8 and MHC-II. Prior to flow cytometry, cells were incubated with propidium iodide (PI; 1 mg/ml) for gating of live (PI−) cells. C57BL/6J splenocytes were stained as a CD45.1− control. (A) Individual spleen APC subsets (L-DC, myeloid cells, CD8− cDC and CD8+ cDC) were gated as shown in Figure 1A and assessed for donor-cell (CD45.1+) composition. Numbers in gates indicate per cent positive cells. Labelled chimeras (*) indicate mice showing only partial donor reconstitution with some evidence of host-type cell development. (B) The relative prevalence of donor- to host-type cells was calculated for each spleen APC subset in animals showing partial donor reconstitution as indicated in Figures 1 and 2 (*). The fold-increase of donor- to host-type cells was calculated relative to CD8− cDC assigned a fold-increase of 1.0. Data represent mean 6 SE (n= 4). Cell subsets having significantly higher representation of donor- over host-type cells, compared with CD8− cDC, are indicated (*) (P≤ 0.02; Wilcoxon Rank Sum Test).
Mentions: The superior reconstitution potential of HSC in neonatal spleen was long-lasting, providing CD45.1+ donor-type haematopoietic cells in all SPL chimeras tested (3 of 3) for more than 25 weeks (Fig. 2A). As with short-term reconstitution analyses shown in Figure 1B, all long-term SPL chimeras gave almost complete donor (CD45.1+) cell reconstitution. This demonstrates that donor-type neonatal spleen cells (CD45.1+) out-compete an equal number of host-type BM cells (CD45.1−) for haematopoietic reconstitution, under the adoptive transfer protocol used here. Complete (.99%) reconstitution of spleen APC subsets by donor-type spleen HSC was observed in two out of three SPL chimeras analysed after 25 weeks. Only one chimera displayed partial donor reconstitution of APC subsets in spleen at 51 weeks, with proportions of subsets varying from 67% to 96%.

Bottom Line: The ability of HSC in spleen to develop into L-DC was indicated by a strong bias in the subset size of these cells over other splenic APC subsets.This type of evidence supports a model whereby spleen represents an important site for haematopoiesis of this distinct DC subset.The conditions under which haematopoiesis of L-DC occurs in spleen, or the progenitors involved, will require further investigation.

View Article: PubMed Central - PubMed

Affiliation: Research School of Biology, The Australian National University, Canberra, Australia.

Show MeSH
Related in: MedlinePlus