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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

The effect of OP9 and OP9DL1 on proliferation of mature B cells. The B cells (B220+) isolated from mice spleens were stained with CFSE, then exposed with 25 ng/mL IL4 plus 10 μg/mL LPS for 24 h, and subsequently cultured alone or together with OP9 or OP9DL1 cells at different rates for 36 h. At 36 h later, the morphology of B cells in each well was observed microscopically (a), then the B cells were analyzed by FACS, and proliferation was measured by the reduction in CFSE intensity. Cell growth rates are indicated by percentages in (b). Data are representative of three experiments.
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Related In: Results  -  Collection


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fig4: The effect of OP9 and OP9DL1 on proliferation of mature B cells. The B cells (B220+) isolated from mice spleens were stained with CFSE, then exposed with 25 ng/mL IL4 plus 10 μg/mL LPS for 24 h, and subsequently cultured alone or together with OP9 or OP9DL1 cells at different rates for 36 h. At 36 h later, the morphology of B cells in each well was observed microscopically (a), then the B cells were analyzed by FACS, and proliferation was measured by the reduction in CFSE intensity. Cell growth rates are indicated by percentages in (b). Data are representative of three experiments.

Mentions: The impact of OP9 or OP9DL1 on mature B cell proliferation has yet to be defined. Splenic B220+ B cells were stained with CFSE and then incubated alone or together with OP9 or OP9DL1 at different ratios (OP9 or OP9DL1 versus B cells) in the presence of LPS plus IL4, respectively. As shown in Figure 4, both OP9 and OP9DL1 inhibit mature B cell proliferation as indicated by the reduction in CFSE intensity. Compared with OP9DL1, OP9 exhibited a much stronger immunosuppressive activity, and B cell proliferation was strikingly inhibited by OP9 at ratios as low as 1 : 80 (OP9 to B).


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)

The effect of OP9 and OP9DL1 on proliferation of mature B cells. The B cells (B220+) isolated from mice spleens were stained with CFSE, then exposed with 25 ng/mL IL4 plus 10 μg/mL LPS for 24 h, and subsequently cultured alone or together with OP9 or OP9DL1 cells at different rates for 36 h. At 36 h later, the morphology of B cells in each well was observed microscopically (a), then the B cells were analyzed by FACS, and proliferation was measured by the reduction in CFSE intensity. Cell growth rates are indicated by percentages in (b). Data are representative of three experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: The effect of OP9 and OP9DL1 on proliferation of mature B cells. The B cells (B220+) isolated from mice spleens were stained with CFSE, then exposed with 25 ng/mL IL4 plus 10 μg/mL LPS for 24 h, and subsequently cultured alone or together with OP9 or OP9DL1 cells at different rates for 36 h. At 36 h later, the morphology of B cells in each well was observed microscopically (a), then the B cells were analyzed by FACS, and proliferation was measured by the reduction in CFSE intensity. Cell growth rates are indicated by percentages in (b). Data are representative of three experiments.
Mentions: The impact of OP9 or OP9DL1 on mature B cell proliferation has yet to be defined. Splenic B220+ B cells were stained with CFSE and then incubated alone or together with OP9 or OP9DL1 at different ratios (OP9 or OP9DL1 versus B cells) in the presence of LPS plus IL4, respectively. As shown in Figure 4, both OP9 and OP9DL1 inhibit mature B cell proliferation as indicated by the reduction in CFSE intensity. Compared with OP9DL1, OP9 exhibited a much stronger immunosuppressive activity, and B cell proliferation was strikingly inhibited by OP9 at ratios as low as 1 : 80 (OP9 to B).

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