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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 suppress monocyte-derived DC differentiation and DC ability to stimulate T-cell proliferation. (a) Mean percentages ± SD of the expression of CD14, CD83, CD40, CD80, CD86, HLA-DR, CD1a, CD54, and α4 and α5 integrin of DCs differentiated in the presence of HLSCs or MSCs or in the absence of stem cells (CTRL) evaluated by cytofluorimetric analyses. Results were obtained from 8 independent experiments. ANOVA with Dunnet's multicomparison test was performed: ∗p < 0.05 stem cells versus CTRL; ∗∗p < 0.001 stem cells versus CTRL. (b) Mean fluorescence intensity expressed as GEO mean ± SD of CD40, CD1a, CD80, α5 integrin, CD86, HLA-DR, and CD54 of DCs differentiated in the presence of HLSCs or MSCs or in the absence of stem cells (CTRL). Results were obtained from 8 independent experiments. ANOVA with Dunnet's multicomparison test was performed: ∗p < 0.05 stem cells versus CTRL; ∗∗p < 0.001 stem cells versus CTRL. (c) DCs differentiated in the presence of HLSCs or in the absence were plated at cell concentration of 2 × 104 with 1 × 105 T CD3+ lymphocytes. Forty-eight hours later, T-cell proliferation was assessed. Data are expressed as mean ± SD of percent variation of T-cell proliferation in the presence or the absence of DCs differentiated in the presence of HLSCs with respect to T-cell proliferation in the presence of DCs alone (established as 100%). Results were obtained from 5 independent experiments. Data were analysed by Student's t-test (unpaired, 2-tailed); ∗p < 0.05 DC (HLSC)+T versus DC+T.
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fig5: HLSCs suppress monocyte-derived DC differentiation and DC ability to stimulate T-cell proliferation. (a) Mean percentages ± SD of the expression of CD14, CD83, CD40, CD80, CD86, HLA-DR, CD1a, CD54, and α4 and α5 integrin of DCs differentiated in the presence of HLSCs or MSCs or in the absence of stem cells (CTRL) evaluated by cytofluorimetric analyses. Results were obtained from 8 independent experiments. ANOVA with Dunnet's multicomparison test was performed: ∗p < 0.05 stem cells versus CTRL; ∗∗p < 0.001 stem cells versus CTRL. (b) Mean fluorescence intensity expressed as GEO mean ± SD of CD40, CD1a, CD80, α5 integrin, CD86, HLA-DR, and CD54 of DCs differentiated in the presence of HLSCs or MSCs or in the absence of stem cells (CTRL). Results were obtained from 8 independent experiments. ANOVA with Dunnet's multicomparison test was performed: ∗p < 0.05 stem cells versus CTRL; ∗∗p < 0.001 stem cells versus CTRL. (c) DCs differentiated in the presence of HLSCs or in the absence were plated at cell concentration of 2 × 104 with 1 × 105 T CD3+ lymphocytes. Forty-eight hours later, T-cell proliferation was assessed. Data are expressed as mean ± SD of percent variation of T-cell proliferation in the presence or the absence of DCs differentiated in the presence of HLSCs with respect to T-cell proliferation in the presence of DCs alone (established as 100%). Results were obtained from 5 independent experiments. Data were analysed by Student's t-test (unpaired, 2-tailed); ∗p < 0.05 DC (HLSC)+T versus DC+T.

Mentions: Monocytes were cultured with GM-CSF and IL-4 in the presence or the absence of HLSCs or MSCs. After 4 days of culture, LPS was added to promote the complete differentiation of DCs. After 7 days of differentiation in these culture conditions, monocytes acquired the characteristic shape and morphology of DCs and their degrees of differentiation and maturation were assessed by FACS analyses. In DCs that differentiated in the absence of HLSCs and MSCs, the expression of CD14 (6 ± 4.8% of positive cells) surface marker was almost abrogated; CD80, CD86, and HLA-DR expression was high (72 ± 10%, 87 ± 8.6%, and 93 ± 6.9%, resp.) and the activation markers CD1a and CD40 were increased (87 ± 6.4%, 23 ± 15.0%, resp.) (Supplementary Figure 3 and Figure 5(a)). The presence of HLSCs cultured in transwells interfered the differentiation process of DCs, as shown by the persistence of CD14 (58 ± 10.5%) expression on monocyte-derived cells (Supplementary Figure 3 and Figure 5(a)) and inhibition of the differentiation and maturation markers such as CD80 (38 ± 16.1%), CD86 (37 ± 20.3%), HLA-DR (51 ± 21.3%), CD1a (45 ± 16.0%), and CD40 (2.8 ± 2.0%). Moreover, the expression of α4 integrin and α5 integrin adhesion molecules and CD54 decreased in the presence of HLSCs (Supplementary Figure 3 and Figure 5(a)). The mean fluorescence intensity (GEO mean) was also analysed for several surface markers. The intensity for CD80, CD86 and HLA-DR, CD54, and α5 integrin was significantly lower in DCs cultured with HLSCs compared with DCs differentiated in the absence of stem cells (Figure 5(b)). The effect of HLSCs on DC differentiation was comparable to that previously described for MSCs (Figures 5(a) and 5(b)) [12, 13]. Notably, the presence of HLSCs did not affect DCs viability (not shown) as already demonstrated for MSCs [12].


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 suppress monocyte-derived DC differentiation and DC ability to stimulate T-cell proliferation. (a) Mean percentages ± SD of the expression of CD14, CD83, CD40, CD80, CD86, HLA-DR, CD1a, CD54, and α4 and α5 integrin of DCs differentiated in the presence of HLSCs or MSCs or in the absence of stem cells (CTRL) evaluated by cytofluorimetric analyses. Results were obtained from 8 independent experiments. ANOVA with Dunnet's multicomparison test was performed: ∗p < 0.05 stem cells versus CTRL; ∗∗p < 0.001 stem cells versus CTRL. (b) Mean fluorescence intensity expressed as GEO mean ± SD of CD40, CD1a, CD80, α5 integrin, CD86, HLA-DR, and CD54 of DCs differentiated in the presence of HLSCs or MSCs or in the absence of stem cells (CTRL). Results were obtained from 8 independent experiments. ANOVA with Dunnet's multicomparison test was performed: ∗p < 0.05 stem cells versus CTRL; ∗∗p < 0.001 stem cells versus CTRL. (c) DCs differentiated in the presence of HLSCs or in the absence were plated at cell concentration of 2 × 104 with 1 × 105 T CD3+ lymphocytes. Forty-eight hours later, T-cell proliferation was assessed. Data are expressed as mean ± SD of percent variation of T-cell proliferation in the presence or the absence of DCs differentiated in the presence of HLSCs with respect to T-cell proliferation in the presence of DCs alone (established as 100%). Results were obtained from 5 independent experiments. Data were analysed by Student's t-test (unpaired, 2-tailed); ∗p < 0.05 DC (HLSC)+T versus DC+T.
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Related In: Results  -  Collection

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fig5: HLSCs suppress monocyte-derived DC differentiation and DC ability to stimulate T-cell proliferation. (a) Mean percentages ± SD of the expression of CD14, CD83, CD40, CD80, CD86, HLA-DR, CD1a, CD54, and α4 and α5 integrin of DCs differentiated in the presence of HLSCs or MSCs or in the absence of stem cells (CTRL) evaluated by cytofluorimetric analyses. Results were obtained from 8 independent experiments. ANOVA with Dunnet's multicomparison test was performed: ∗p < 0.05 stem cells versus CTRL; ∗∗p < 0.001 stem cells versus CTRL. (b) Mean fluorescence intensity expressed as GEO mean ± SD of CD40, CD1a, CD80, α5 integrin, CD86, HLA-DR, and CD54 of DCs differentiated in the presence of HLSCs or MSCs or in the absence of stem cells (CTRL). Results were obtained from 8 independent experiments. ANOVA with Dunnet's multicomparison test was performed: ∗p < 0.05 stem cells versus CTRL; ∗∗p < 0.001 stem cells versus CTRL. (c) DCs differentiated in the presence of HLSCs or in the absence were plated at cell concentration of 2 × 104 with 1 × 105 T CD3+ lymphocytes. Forty-eight hours later, T-cell proliferation was assessed. Data are expressed as mean ± SD of percent variation of T-cell proliferation in the presence or the absence of DCs differentiated in the presence of HLSCs with respect to T-cell proliferation in the presence of DCs alone (established as 100%). Results were obtained from 5 independent experiments. Data were analysed by Student's t-test (unpaired, 2-tailed); ∗p < 0.05 DC (HLSC)+T versus DC+T.
Mentions: Monocytes were cultured with GM-CSF and IL-4 in the presence or the absence of HLSCs or MSCs. After 4 days of culture, LPS was added to promote the complete differentiation of DCs. After 7 days of differentiation in these culture conditions, monocytes acquired the characteristic shape and morphology of DCs and their degrees of differentiation and maturation were assessed by FACS analyses. In DCs that differentiated in the absence of HLSCs and MSCs, the expression of CD14 (6 ± 4.8% of positive cells) surface marker was almost abrogated; CD80, CD86, and HLA-DR expression was high (72 ± 10%, 87 ± 8.6%, and 93 ± 6.9%, resp.) and the activation markers CD1a and CD40 were increased (87 ± 6.4%, 23 ± 15.0%, resp.) (Supplementary Figure 3 and Figure 5(a)). The presence of HLSCs cultured in transwells interfered the differentiation process of DCs, as shown by the persistence of CD14 (58 ± 10.5%) expression on monocyte-derived cells (Supplementary Figure 3 and Figure 5(a)) and inhibition of the differentiation and maturation markers such as CD80 (38 ± 16.1%), CD86 (37 ± 20.3%), HLA-DR (51 ± 21.3%), CD1a (45 ± 16.0%), and CD40 (2.8 ± 2.0%). Moreover, the expression of α4 integrin and α5 integrin adhesion molecules and CD54 decreased in the presence of HLSCs (Supplementary Figure 3 and Figure 5(a)). The mean fluorescence intensity (GEO mean) was also analysed for several surface markers. The intensity for CD80, CD86 and HLA-DR, CD54, and α5 integrin was significantly lower in DCs cultured with HLSCs compared with DCs differentiated in the absence of stem cells (Figure 5(b)). The effect of HLSCs on DC differentiation was comparable to that previously described for MSCs (Figures 5(a) and 5(b)) [12, 13]. Notably, the presence of HLSCs did not affect DCs viability (not shown) as already demonstrated for MSCs [12].

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