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Human Liver Stem Cells Suppress T-Cell Proliferation, NK Activity, and Dendritic Cell Differentiation.

Bruno S, Grange C, Tapparo M, Pasquino C, Romagnoli R, Dametto E, Amoroso A, Tetta C, Camussi G - Stem Cells Int (2016)

Bottom Line: At variance with MSCs, HLSCs did not elicit NK degranulation.Moreover, HLSCs inhibited NK degranulation against K562, a NK-sensitive target, by a mechanism dependent on HLA-G release.This study shows that HLSCs have immunomodulatory properties similar to MSCs, but, at variance with MSCs, they do not elicit a NK response.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biotechnology and Health Science, University of Torino, 10126 Torino, Italy.

ABSTRACT
Human liver stem cells (HLSCs) are a mesenchymal stromal cell-like population resident in the adult liver. Preclinical studies indicate that HLSCs could be a good candidate for cell therapy. The aim of the present study was to evaluate the immunogenicity and the immunomodulatory properties of HLSCs on T-lymphocytes, natural killer cells (NKs), and dendritic cells (DCs) in allogeneic experimental settings. We found that HLSCs inhibited T-cell proliferation by a mechanism independent of cell contact and dependent on the release of prostaglandin E2 (PGE2) and on indoleamine 2,3-dioxygenase activity. When compared with mesenchymal stromal cells (MSCs), HLSCs were more efficient in inhibiting T-cell proliferation. At variance with MSCs, HLSCs did not elicit NK degranulation. Moreover, HLSCs inhibited NK degranulation against K562, a NK-sensitive target, by a mechanism dependent on HLA-G release. When tested on DC generation from monocytes, HLSCs were found to impair DC differentiation and DCs ability to induce T-cell proliferation through PGE2. This study shows that HLSCs have immunomodulatory properties similar to MSCs, but, at variance with MSCs, they do not elicit a NK response.

No MeSH data available.


Related in: MedlinePlus

HLSCs inhibit NK activity. NKs, prestimulated for 24 hours with IL-15, were cocultured at the ratio of 5 : 1 with MSCs or HLSCs in direct contact or in transwells for 4 days. Subsequently, the NKs were challenged with a NK-susceptible target cell line (K562 cell line) and their degranulation was evaluated. (a) Representative (n = 3) cytofluorimetric analysis of CD107a expression on NKs (NK CTRL) and on NKs after 4 hours of incubation with K562 cells (NK CTRL+K562). CD107a was detectable on NK membrane after incubation with K562 cells. (b) Representative (n = 5) cytofluorimetric analysis of CD107a expression on NKs in the presence of K562 and after 4 days of incubation with MSCs or HLSCs. NKs cocultured with MSCs and HLSCs reduced their capacity to degranulate in the presence of specific target cells. (c)–(e) The death of K562 cells labelled with CFSE, which were evaluated by PI staining, after incubation for 4 hours with NKs stimulated with IL-15 (c) and cocultured, in the presence of transwells, for 4 days with MSCs (d) or with HLSCs (e).
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fig3: HLSCs inhibit NK activity. NKs, prestimulated for 24 hours with IL-15, were cocultured at the ratio of 5 : 1 with MSCs or HLSCs in direct contact or in transwells for 4 days. Subsequently, the NKs were challenged with a NK-susceptible target cell line (K562 cell line) and their degranulation was evaluated. (a) Representative (n = 3) cytofluorimetric analysis of CD107a expression on NKs (NK CTRL) and on NKs after 4 hours of incubation with K562 cells (NK CTRL+K562). CD107a was detectable on NK membrane after incubation with K562 cells. (b) Representative (n = 5) cytofluorimetric analysis of CD107a expression on NKs in the presence of K562 and after 4 days of incubation with MSCs or HLSCs. NKs cocultured with MSCs and HLSCs reduced their capacity to degranulate in the presence of specific target cells. (c)–(e) The death of K562 cells labelled with CFSE, which were evaluated by PI staining, after incubation for 4 hours with NKs stimulated with IL-15 (c) and cocultured, in the presence of transwells, for 4 days with MSCs (d) or with HLSCs (e).

Mentions: To study whether HLSCs modulated NK activity, coculture experiments were carried out in direct contact or by using transwell inserts. Prestimulated NKs were cocultured with HLSCs or MSCs and subsequently harvested to evaluate the degranulation capacity against K562, a susceptible target cell line. K562 cells induced degranulation of NKs prestimulated with IL-15 (CD56+CD107+: 65 ± 8% (Figure 3(a))). Preincubation of NKs with HLSCs, in contact condition or in the presence of transwell, was able to significantly decrease NK degranulation (CD56+CD107+: resp., 8.2 ± 1.5% and 2.5 ± 2%), in a manner comparable with the preincubation with MSCs (CD56+CD107+: resp., 5.3 ± 2.2% and 7 ± 3%) (Figure 3(b)). Moreover, after incubation with HLSCs, NKs decreased their capacity to kill K562 cells (Figures 3(c) and 3(e)) and confirmed the data obtained by the degranulation test. Also NKs incubated with MSCs decreased their capacity to kill K562 cells (Figures 3(c) and 3(d)), but to a lesser extent than HLSCs (Figure 3(e)).


Human Liver Stem Cells Suppress T-Cell Proliferation, NK Activity, and Dendritic Cell Differentiation.

Bruno S, Grange C, Tapparo M, Pasquino C, Romagnoli R, Dametto E, Amoroso A, Tetta C, Camussi G - Stem Cells Int (2016)

HLSCs inhibit NK activity. NKs, prestimulated for 24 hours with IL-15, were cocultured at the ratio of 5 : 1 with MSCs or HLSCs in direct contact or in transwells for 4 days. Subsequently, the NKs were challenged with a NK-susceptible target cell line (K562 cell line) and their degranulation was evaluated. (a) Representative (n = 3) cytofluorimetric analysis of CD107a expression on NKs (NK CTRL) and on NKs after 4 hours of incubation with K562 cells (NK CTRL+K562). CD107a was detectable on NK membrane after incubation with K562 cells. (b) Representative (n = 5) cytofluorimetric analysis of CD107a expression on NKs in the presence of K562 and after 4 days of incubation with MSCs or HLSCs. NKs cocultured with MSCs and HLSCs reduced their capacity to degranulate in the presence of specific target cells. (c)–(e) The death of K562 cells labelled with CFSE, which were evaluated by PI staining, after incubation for 4 hours with NKs stimulated with IL-15 (c) and cocultured, in the presence of transwells, for 4 days with MSCs (d) or with HLSCs (e).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: HLSCs inhibit NK activity. NKs, prestimulated for 24 hours with IL-15, were cocultured at the ratio of 5 : 1 with MSCs or HLSCs in direct contact or in transwells for 4 days. Subsequently, the NKs were challenged with a NK-susceptible target cell line (K562 cell line) and their degranulation was evaluated. (a) Representative (n = 3) cytofluorimetric analysis of CD107a expression on NKs (NK CTRL) and on NKs after 4 hours of incubation with K562 cells (NK CTRL+K562). CD107a was detectable on NK membrane after incubation with K562 cells. (b) Representative (n = 5) cytofluorimetric analysis of CD107a expression on NKs in the presence of K562 and after 4 days of incubation with MSCs or HLSCs. NKs cocultured with MSCs and HLSCs reduced their capacity to degranulate in the presence of specific target cells. (c)–(e) The death of K562 cells labelled with CFSE, which were evaluated by PI staining, after incubation for 4 hours with NKs stimulated with IL-15 (c) and cocultured, in the presence of transwells, for 4 days with MSCs (d) or with HLSCs (e).
Mentions: To study whether HLSCs modulated NK activity, coculture experiments were carried out in direct contact or by using transwell inserts. Prestimulated NKs were cocultured with HLSCs or MSCs and subsequently harvested to evaluate the degranulation capacity against K562, a susceptible target cell line. K562 cells induced degranulation of NKs prestimulated with IL-15 (CD56+CD107+: 65 ± 8% (Figure 3(a))). Preincubation of NKs with HLSCs, in contact condition or in the presence of transwell, was able to significantly decrease NK degranulation (CD56+CD107+: resp., 8.2 ± 1.5% and 2.5 ± 2%), in a manner comparable with the preincubation with MSCs (CD56+CD107+: resp., 5.3 ± 2.2% and 7 ± 3%) (Figure 3(b)). Moreover, after incubation with HLSCs, NKs decreased their capacity to kill K562 cells (Figures 3(c) and 3(e)) and confirmed the data obtained by the degranulation test. Also NKs incubated with MSCs decreased their capacity to kill K562 cells (Figures 3(c) and 3(d)), but to a lesser extent than HLSCs (Figure 3(e)).

Bottom Line: At variance with MSCs, HLSCs did not elicit NK degranulation.Moreover, HLSCs inhibited NK degranulation against K562, a NK-sensitive target, by a mechanism dependent on HLA-G release.This study shows that HLSCs have immunomodulatory properties similar to MSCs, but, at variance with MSCs, they do not elicit a NK response.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biotechnology and Health Science, University of Torino, 10126 Torino, Italy.

ABSTRACT
Human liver stem cells (HLSCs) are a mesenchymal stromal cell-like population resident in the adult liver. Preclinical studies indicate that HLSCs could be a good candidate for cell therapy. The aim of the present study was to evaluate the immunogenicity and the immunomodulatory properties of HLSCs on T-lymphocytes, natural killer cells (NKs), and dendritic cells (DCs) in allogeneic experimental settings. We found that HLSCs inhibited T-cell proliferation by a mechanism independent of cell contact and dependent on the release of prostaglandin E2 (PGE2) and on indoleamine 2,3-dioxygenase activity. When compared with mesenchymal stromal cells (MSCs), HLSCs were more efficient in inhibiting T-cell proliferation. At variance with MSCs, HLSCs did not elicit NK degranulation. Moreover, HLSCs inhibited NK degranulation against K562, a NK-sensitive target, by a mechanism dependent on HLA-G release. When tested on DC generation from monocytes, HLSCs were found to impair DC differentiation and DCs ability to induce T-cell proliferation through PGE2. This study shows that HLSCs have immunomodulatory properties similar to MSCs, but, at variance with MSCs, they do not elicit a NK response.

No MeSH data available.


Related in: MedlinePlus