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Delta-Like-1 Changes the Immunomodulatory Property of OP9 Cells.

Zhang L, Dang RJ, Yang YM, Cui DC, Li P, Ni YL, Hao T, Wang C, Jiang XX, Fang NZ - Stem Cells Int (2015)

Bottom Line: With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production.The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγ or LPS/IL4.DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Animal Genetic Breeding and Reproduction, Yanbian University, Yanji, Jilin 133002, China ; Department of Advanced Interdisciplinary Studies, Institute of Basic Medical Sciences, 27 Taiping Road, Haidian District, Beijing 100850, China ; Department of Biology and Chemical Engineering, Tongren University, Tongren, Guizhou 554300, China.

ABSTRACT
As stromal cells and recently confirmed mesenchymal stem cells, OP9 cells support hematopoiesis stem cell (HSC) differentiation into the B lymphocyte lineage, yet Delta-like-1 (DL1) overexpressing OP9 (OP9DL1) cells promote the development of early T lymphocytes from HSC. However, the immunomodulatory capacity of OP9 or OP9DL1 on mature B and T cell proliferation has not been elucidated. Here, we show that OP9 and OP9DL1 have similar proliferation capacities and immunophenotypes except DL1 expression. Compared with OP9, OP9DL1 displayed more osteogenesis and less adipogenesis when cultured in the respective induction media. Both OP9 and OP9DL1 inhibited mature B and T cell proliferation. Furthermore, OP9 showed stronger inhibition on B cell proliferation and OP9DL1 exhibited stronger inhibition on T cell proliferation. With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production. The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγ or LPS/IL4. Taken together, our study reveals a previously unrecognized role of OP9 and OP9DL1 in mature B and T cell proliferation. DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

No MeSH data available.


Related in: MedlinePlus

OP9 and OP9DL1 inhibit proliferation of mature T cells in vitro. CD3+ T cells were isolated from murine spleens with CD3ε MicroBead Kits and labeled with CFSE. T cells were stimulated with PMA (50 ng/mL) plus ionomycin (1 μg/mL) for 24 h and then cultured alone or with OP9 or OP9DL1 at different ratios (OP9 or OP9DL1 cells to T cells). After 36 h, all of the cells were analyzed via microscope (a) and flow cytometry (b) for T cell proliferation as indicated by the reduction in CFSE intensity. Data are representative of three independent experiments.
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fig5: OP9 and OP9DL1 inhibit proliferation of mature T cells in vitro. CD3+ T cells were isolated from murine spleens with CD3ε MicroBead Kits and labeled with CFSE. T cells were stimulated with PMA (50 ng/mL) plus ionomycin (1 μg/mL) for 24 h and then cultured alone or with OP9 or OP9DL1 at different ratios (OP9 or OP9DL1 cells to T cells). After 36 h, all of the cells were analyzed via microscope (a) and flow cytometry (b) for T cell proliferation as indicated by the reduction in CFSE intensity. Data are representative of three independent experiments.

Mentions: As mentioned earlier, OP9DL1 promotes early T cell development, but its effect on mature T cell proliferation is unknown. To clarify the role of OP9DL1 on mature T cell growth, CD3+ T cells stained with CFSE were cultured alone or together with OP9 or OP9DL1 cells at the indicated ratios (Figure 5). Mature T cell proliferation was directly assessed by CFSE labeling and monitoring of CFSE dilution. Unexpectedly, a marked reduction in T cell proliferation in the OP9DL1 cultures was observed compared with that in coculture with OP9 at each proportional hierarchy (Figure 5). Compared with the growth rate of T cells alone, OP9DL1 inhibits T cell proliferation more obviously with increasing ratios (OP9DL1 versus T cell), while OP9 only prohibits T cell growth at the highest ratio.


Delta-Like-1 Changes the Immunomodulatory Property of OP9 Cells.

Zhang L, Dang RJ, Yang YM, Cui DC, Li P, Ni YL, Hao T, Wang C, Jiang XX, Fang NZ - Stem Cells Int (2015)

OP9 and OP9DL1 inhibit proliferation of mature T cells in vitro. CD3+ T cells were isolated from murine spleens with CD3ε MicroBead Kits and labeled with CFSE. T cells were stimulated with PMA (50 ng/mL) plus ionomycin (1 μg/mL) for 24 h and then cultured alone or with OP9 or OP9DL1 at different ratios (OP9 or OP9DL1 cells to T cells). After 36 h, all of the cells were analyzed via microscope (a) and flow cytometry (b) for T cell proliferation as indicated by the reduction in CFSE intensity. Data are representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4663344&req=5

fig5: OP9 and OP9DL1 inhibit proliferation of mature T cells in vitro. CD3+ T cells were isolated from murine spleens with CD3ε MicroBead Kits and labeled with CFSE. T cells were stimulated with PMA (50 ng/mL) plus ionomycin (1 μg/mL) for 24 h and then cultured alone or with OP9 or OP9DL1 at different ratios (OP9 or OP9DL1 cells to T cells). After 36 h, all of the cells were analyzed via microscope (a) and flow cytometry (b) for T cell proliferation as indicated by the reduction in CFSE intensity. Data are representative of three independent experiments.
Mentions: As mentioned earlier, OP9DL1 promotes early T cell development, but its effect on mature T cell proliferation is unknown. To clarify the role of OP9DL1 on mature T cell growth, CD3+ T cells stained with CFSE were cultured alone or together with OP9 or OP9DL1 cells at the indicated ratios (Figure 5). Mature T cell proliferation was directly assessed by CFSE labeling and monitoring of CFSE dilution. Unexpectedly, a marked reduction in T cell proliferation in the OP9DL1 cultures was observed compared with that in coculture with OP9 at each proportional hierarchy (Figure 5). Compared with the growth rate of T cells alone, OP9DL1 inhibits T cell proliferation more obviously with increasing ratios (OP9DL1 versus T cell), while OP9 only prohibits T cell growth at the highest ratio.

Bottom Line: With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production.The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγ or LPS/IL4.DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Animal Genetic Breeding and Reproduction, Yanbian University, Yanji, Jilin 133002, China ; Department of Advanced Interdisciplinary Studies, Institute of Basic Medical Sciences, 27 Taiping Road, Haidian District, Beijing 100850, China ; Department of Biology and Chemical Engineering, Tongren University, Tongren, Guizhou 554300, China.

ABSTRACT
As stromal cells and recently confirmed mesenchymal stem cells, OP9 cells support hematopoiesis stem cell (HSC) differentiation into the B lymphocyte lineage, yet Delta-like-1 (DL1) overexpressing OP9 (OP9DL1) cells promote the development of early T lymphocytes from HSC. However, the immunomodulatory capacity of OP9 or OP9DL1 on mature B and T cell proliferation has not been elucidated. Here, we show that OP9 and OP9DL1 have similar proliferation capacities and immunophenotypes except DL1 expression. Compared with OP9, OP9DL1 displayed more osteogenesis and less adipogenesis when cultured in the respective induction media. Both OP9 and OP9DL1 inhibited mature B and T cell proliferation. Furthermore, OP9 showed stronger inhibition on B cell proliferation and OP9DL1 exhibited stronger inhibition on T cell proliferation. With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production. The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγ or LPS/IL4. Taken together, our study reveals a previously unrecognized role of OP9 and OP9DL1 in mature B and T cell proliferation. DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

No MeSH data available.


Related in: MedlinePlus