Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
, 22 (22), 2990-3002

Comparative Study of Immune Regulatory Properties of Stem Cells Derived From Different Tissues

Affiliations
Comparative Study

Comparative Study of Immune Regulatory Properties of Stem Cells Derived From Different Tissues

Mariano Di Trapani et al. Stem Cells Dev.

Abstract

Allogeneic stem cell (SC)-based therapy is a promising tool for the treatment of a range of human degenerative and inflammatory diseases. Many reports highlighted the immune modulatory properties of some SC types, such as mesenchymal stromal cells (MSCs), but a comparative study with SCs of different origin, to assess whether immune regulation is a general SC property, is still lacking. To this aim, we applied highly standardized methods employed for MSC characterization to compare the immunological properties of bone marrow-MSCs, olfactory ectomesenchymal SCs, leptomeningeal SCs, and three different c-Kit-positive SC types, that is, amniotic fluid SCs, cardiac SCs, and lung SCs. We found that all the analyzed human SCs share a common pattern of immunological features, in terms of expression of activation markers ICAM-1, VCAM-1, HLA-ABC, and HLA-DR, modulatory activity toward purified T, B, and NK cells, lower immunogenicity of inflammatory-primed SCs as compared to resting SCs, and indoleamine-2,3-dioxygenase-activation as molecular inhibitory pathways, with some SC type-related peculiarities. Moreover, the SC types analyzed exert an anti-apoptotic effect toward not-activated immune effector cells (IECs). In addition, we found that the inhibitory behavior is not a constitutive property of SCs, but is acquired as a consequence of IEC activation, as previously described for MSCs. Thus, immune regulation is a general property of SCs and the characterization of this phenomenon may be useful for a proper therapeutic use of SCs.

Figures

FIG. 1.
FIG. 1.
Representative immunofluorescence staining of various human stem cell (SC) types. (A) Bone marrow (BM)-mesenchymal stromal cells (MSCs) were stained with anti-CD73-PE (red) and TOPRO-3 (blue); (B) olfactory ectomesenchymal SCs (OE-MSCs) were stained with anti-Nestin (green) and Hoechst-33342 (blue); (C) leptomeningeal SCs (LeSCs) were stained with anti-Nestin (red) and TOPRO-3 (blue); (D) amniotic fluid SCs (AFSCs) were stained with anti-c-Kit (green) and TOPRO-3 (blue); (E) cardiac SC (CSCs) were stained with anti-c-Kit (white) and TOPRO-3; (F) lung SCs (LSCs) were stained with anti-c-Kit (green) and TOPRO-3 (blue). Scale bars: 50 μm (A–C); 20 μm (D–F).
FIG. 2.
FIG. 2.
Hierarchical cluster analysis of protein expression of various human SCs in resting and primed conditions. Modulation of expression of fifteen proteins in resting and inflammatory-primed conditions. Heat map showed the down- (green) or upregulation (red) of protein expression. Different samples were grouped using hierarchical clustering algorithm.
FIG. 3.
FIG. 3.
Human SC inhibitory effect on stimulated immune effector cell (IEC) proliferation. Human IEC (ie, T, NK, and B cells) were stimulated with specific stimuli and cultured alone (dark gray bars) or in the presence of resting (white bars) or primed (light gray bars) allogeneic human SCs. At the end of coculture, lymphocyte proliferation was assessed using carboxyfluorescein succinimidyl ester (CFSE) dilution method, as described in Materials and Methods section. CFSE fluorescence was analyzed after 7 days for T (at 10:1 T/SC ratio) and NK (at 1:1 NK/SC ratio) cells (A and B, respectively), while for B cells (C) the fluorescence was detected after 4 days of coculture (at 1:1 B/SCs ratio). The results are expressed as relative proliferation percentage of IEC, normalized to IEC cultured alone (100%). Error bars represented mean±SD of five independent experiments for BM-MSCs, OE-MSCs, LeSCs, and AFSCs and three independent experiments for CSCs and LSCs. *P<0.05, **P<0.01, ***P<0.001.
FIG. 4.
FIG. 4.
Effect of specific inhibitors on T cell proliferation. Human purified T cells were stained with CFSE, stimulated with mitogenic anti-CD3 and anti-CD28 antibodies, and cultured alone or in presence of different resting and primed SC types (at 10:1 T/SCs ratio). In each coculture the following inhibitors (I) were added: L-1MT (A), Anti-IFN-γ (B), NS-398 (C), and snPP (D). After 6 days, cells were harvested and T cell proliferation was evaluated by FACS analysis. The results are expressed as relative proliferation percentage of T cells, normalized to T cells cultured alone (100%). Error bars represented mean±SD of five independent experiments for BM-MSCs, OE-MSCs, LeSCs, and AFSCs and three independent experiments for CSCs and LSCs. *P<0.05, **P<0.01, ***P<0.001.
FIG. 5.
FIG. 5.
Immunogenicity of resting and primed human SCs. To evaluate NK cytotoxicity, SCs were labeled with BATDA (as reported in Materials and Methods section) and then used as effector cells in coculture with IL-2-stimulated NK cells at different NK/SC ratios. The amount of cytotoxicity was calculated as release of fluorescence by lysed SCs, and detected by time-resolved fluorimeter (Victor X4 Multilabel Plate Reader, PerkinElmer). Each graphic shows the results obtained from five independent experiments, in which resting (solid line) and primed (dashed line) SCs were used as target cells. Data are expressed as percentage of fluorescence release. Error bars represented mean±SD of five independent experiments for BM-MSCs, OE-MSCs, LeSCs, and AFSCs and three independent experiments for CSCs and LSCs. *P<0.05, **P<0.01, ***P<0.001. BATDA, bis-acetoxymethyl terpyridine dicarboxylate.
FIG. 6.
FIG. 6.
Trophic support of resting and primed human SCs on different immune effectors. Resting and primed SCs were cocultured with unstimulated human IECs (ie, T, B, and NK cells). At the end of coculture, immune cell survival was detected by measuring cytosolic active-caspase-3 (as reported in Materials and Methods section). The results are expressed as percentage of caspase-3negCD45pos cells, and the analysis was performed by flow cytometry. Error bars represented mean±SD of five independent experiments for BM-MSCs, OE-MSCs, LeSCs, and AFSCs and three independent experiments for CSCs and LSCs. *P<0.05, **P<0.01, ***P<0.001.

Similar articles

See all similar articles

Cited by 27 articles

See all "Cited by" articles

Publication types

MeSH terms

LinkOut - more resources

Feedback