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Calcineurin/NFAT signalling inhibits myeloid haematopoiesis.

Fric J, Lim CX, Koh EG, Hofmann B, Chen J, Tay HS, Mohammad Isa SA, Mortellaro A, Ruedl C, Ricciardi-Castagnoli P - EMBO Mol Med (2012)

Bottom Line: Reconstituting lethally irradiated mice with haematopoietic stem cells expressing an NFAT-inhibitory peptide resulted in enhanced development of the myeloid compartment.Global gene expression analysis of untreated DC and NFAT-inhibited DC revealed differential expression of transcripts that regulate cell cycle and apoptosis.In conclusion, these results provide evidence that calcineurin/NFAT signalling negatively regulates myeloid lineage development.

View Article: PubMed Central - PubMed

Affiliation: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Singapore, Singapore.

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Related in: MedlinePlus

Mouse reconstitution with primary BM cells expressing VIVIT recapitulates myeloid cell growth advantageFreshly isolated BM cells were transduced with VIVIT-eGFP or tdTomato control, and used for reconstitution of lethally irradiated mice. The proportions of engrafted BM cells were 30% VIVIT-eGFP expressing and 70% control Tomato. Changes in the original ratio in BM and spleen were analysed 4, 8 and 12 weeks after reconstitution (n = 4–5). The initial ratio of HSC used for reconstitution is shown. The ratio between VIVIT-eGFP and control tdTomato cells in each lineage is plotted as the logarithm of the percentage of competitor VIVIT-eGFP cells, divided by percentage of control tdTomato cells (value 0.36 reflects 30:70 ratio). Data show one representative experiment of three. Changes in the ratios between two-time points have been analysed by Student t-test *p ≤ 0.05.
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fig03: Mouse reconstitution with primary BM cells expressing VIVIT recapitulates myeloid cell growth advantageFreshly isolated BM cells were transduced with VIVIT-eGFP or tdTomato control, and used for reconstitution of lethally irradiated mice. The proportions of engrafted BM cells were 30% VIVIT-eGFP expressing and 70% control Tomato. Changes in the original ratio in BM and spleen were analysed 4, 8 and 12 weeks after reconstitution (n = 4–5). The initial ratio of HSC used for reconstitution is shown. The ratio between VIVIT-eGFP and control tdTomato cells in each lineage is plotted as the logarithm of the percentage of competitor VIVIT-eGFP cells, divided by percentage of control tdTomato cells (value 0.36 reflects 30:70 ratio). Data show one representative experiment of three. Changes in the ratios between two-time points have been analysed by Student t-test *p ≤ 0.05.

Mentions: In further studies, these data were confirmed using freshly transduced primary BM cells to replicate the results obtained with HSC lines. Freshly isolated BM cells were transduced with either VIVIT-eGFP or control tdTomato constructs before the cells were sorted and then injected into irradiated hosts in the ratio 30% VIVIT-eGFP competitors to 70% control tdTomato cells. Under these conditions using primary cells and excluding any possible interference of NUP-HOXB4 constructs, we still observed a preferential expansion of calcineurin/NFAT-impaired myeloid cells over controls. The ratio of granulocytes, DC and T cells was analysed at weeks 4–8 and 12 weeks post-engraftment. The competitive advantage of VIVIT-expressing myeloid cells over the mock-transduced controls was still evident (Fig 3).


Calcineurin/NFAT signalling inhibits myeloid haematopoiesis.

Fric J, Lim CX, Koh EG, Hofmann B, Chen J, Tay HS, Mohammad Isa SA, Mortellaro A, Ruedl C, Ricciardi-Castagnoli P - EMBO Mol Med (2012)

Mouse reconstitution with primary BM cells expressing VIVIT recapitulates myeloid cell growth advantageFreshly isolated BM cells were transduced with VIVIT-eGFP or tdTomato control, and used for reconstitution of lethally irradiated mice. The proportions of engrafted BM cells were 30% VIVIT-eGFP expressing and 70% control Tomato. Changes in the original ratio in BM and spleen were analysed 4, 8 and 12 weeks after reconstitution (n = 4–5). The initial ratio of HSC used for reconstitution is shown. The ratio between VIVIT-eGFP and control tdTomato cells in each lineage is plotted as the logarithm of the percentage of competitor VIVIT-eGFP cells, divided by percentage of control tdTomato cells (value 0.36 reflects 30:70 ratio). Data show one representative experiment of three. Changes in the ratios between two-time points have been analysed by Student t-test *p ≤ 0.05.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3376854&req=5

fig03: Mouse reconstitution with primary BM cells expressing VIVIT recapitulates myeloid cell growth advantageFreshly isolated BM cells were transduced with VIVIT-eGFP or tdTomato control, and used for reconstitution of lethally irradiated mice. The proportions of engrafted BM cells were 30% VIVIT-eGFP expressing and 70% control Tomato. Changes in the original ratio in BM and spleen were analysed 4, 8 and 12 weeks after reconstitution (n = 4–5). The initial ratio of HSC used for reconstitution is shown. The ratio between VIVIT-eGFP and control tdTomato cells in each lineage is plotted as the logarithm of the percentage of competitor VIVIT-eGFP cells, divided by percentage of control tdTomato cells (value 0.36 reflects 30:70 ratio). Data show one representative experiment of three. Changes in the ratios between two-time points have been analysed by Student t-test *p ≤ 0.05.
Mentions: In further studies, these data were confirmed using freshly transduced primary BM cells to replicate the results obtained with HSC lines. Freshly isolated BM cells were transduced with either VIVIT-eGFP or control tdTomato constructs before the cells were sorted and then injected into irradiated hosts in the ratio 30% VIVIT-eGFP competitors to 70% control tdTomato cells. Under these conditions using primary cells and excluding any possible interference of NUP-HOXB4 constructs, we still observed a preferential expansion of calcineurin/NFAT-impaired myeloid cells over controls. The ratio of granulocytes, DC and T cells was analysed at weeks 4–8 and 12 weeks post-engraftment. The competitive advantage of VIVIT-expressing myeloid cells over the mock-transduced controls was still evident (Fig 3).

Bottom Line: Reconstituting lethally irradiated mice with haematopoietic stem cells expressing an NFAT-inhibitory peptide resulted in enhanced development of the myeloid compartment.Global gene expression analysis of untreated DC and NFAT-inhibited DC revealed differential expression of transcripts that regulate cell cycle and apoptosis.In conclusion, these results provide evidence that calcineurin/NFAT signalling negatively regulates myeloid lineage development.

View Article: PubMed Central - PubMed

Affiliation: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Singapore, Singapore.

Show MeSH
Related in: MedlinePlus