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Osteodifferentiated mesenchymal stem cells from bone marrow and adipose tissue express HLA-G and display immunomodulatory properties in HLA-mismatched settings: implications in bone repair therapy.

Montespan F, Deschaseaux F, Sensébé L, Carosella ED, Rouas-Freiss N - J Immunol Res (2014)

Bottom Line: Both MSCs and osteodifferentiated MSCs are hypoimmunogenic and exert immunomodulatory properties in HLA-mismatched settings as they suppress T cell alloproliferation in mixed lymphocyte reactions.Finally, addition of biomaterials that stimulate bone tissue formation did not modify MSC immune properties.As MSCs combine both abilities of osteoregeneration and immunomodulation, they may be considered as allogenic sources for the treatment of bone defects.

View Article: PubMed Central - PubMed

Affiliation: CEA, Institut des Maladies Emergentes et des Therapies Innovantes (IMETI), Service de Recherche en Hemato-Immunologie (SRHI), Hopital Saint-Louis, IUH, 1 avenue Claude Vellefaux, 75010 Paris, France ; Universite Paris Diderot, Sorbonne Paris Cité, IUH, Hopital Saint-Louis, UMR_E5, IUH, 1, avenue Claude Vellefaux, 75010 Paris, France.

ABSTRACT
Mesenchymal stem cells (MSCs) are multipotent cells that can be obtained from several sources such as bone marrow and adipose tissue. Depending on the culture conditions, they can differentiate into osteoblasts, chondroblasts, adipocytes, or neurons. In this regard, they constitute promising candidates for cell-based therapy aimed at repairing damaged tissues. In addition, MSCs display immunomodulatory properties through the expression of soluble factors including HLA-G. We here analyse both immunogenicity and immunosuppressive capacity of MSCs derived from bone marrow and adipose tissue before and after osteodifferentiation. Results show that HLA-G expression is maintained after osteodifferentiation and can be boosted in inflammatory conditions mimicked by the addition of IFN-γ and TNF-α. Both MSCs and osteodifferentiated MSCs are hypoimmunogenic and exert immunomodulatory properties in HLA-mismatched settings as they suppress T cell alloproliferation in mixed lymphocyte reactions. Finally, addition of biomaterials that stimulate bone tissue formation did not modify MSC immune properties. As MSCs combine both abilities of osteoregeneration and immunomodulation, they may be considered as allogenic sources for the treatment of bone defects.

Show MeSH
BM-derived MSCs committed to osteodifferentiation process express HLA-G and are hypoimmunogenic. (a and b) Expression of ALP and HLA-G5 was evaluated by intracellular flow cytometry analysis on BM-derived MSCs committed to 14-day osteodifferentiation process and pretreated with IFNγ and TNFα (Os-MSC-BM IFNγ/TNFα) or not (Os-MSC-BM). IgG1 was used as isotype control Ab. Tubulin was used as positive control of cell permeabilization. (c) PBMC from healthy individual (#00) were used as responder cells towards BM-derived MSCs committed to osteodifferentiation process and pretreated with IFNγ and TNFα (Os-MSC-BM IFNγ/TNFα) or not (Os-MSC-BM) as stimulating cells at various responder: stimulator ratios. Irradiated LCL* were used as positive control of T cell alloproliferation. Results are given as mean cpm ± s.e.m.; one representative experiment is shown.
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fig3: BM-derived MSCs committed to osteodifferentiation process express HLA-G and are hypoimmunogenic. (a and b) Expression of ALP and HLA-G5 was evaluated by intracellular flow cytometry analysis on BM-derived MSCs committed to 14-day osteodifferentiation process and pretreated with IFNγ and TNFα (Os-MSC-BM IFNγ/TNFα) or not (Os-MSC-BM). IgG1 was used as isotype control Ab. Tubulin was used as positive control of cell permeabilization. (c) PBMC from healthy individual (#00) were used as responder cells towards BM-derived MSCs committed to osteodifferentiation process and pretreated with IFNγ and TNFα (Os-MSC-BM IFNγ/TNFα) or not (Os-MSC-BM) as stimulating cells at various responder: stimulator ratios. Irradiated LCL* were used as positive control of T cell alloproliferation. Results are given as mean cpm ± s.e.m.; one representative experiment is shown.

Mentions: No PBMC alloproliferation was observed in response to various doses of allogenic MSCs derived from bone marrow (Figure 1(c)) or adipose tissue (Figure 2(c)) even after licensing with IFN-γ and TNF-α (Figures 1(c) and 2(c)). Tables 1 and 2 summarize the results obtained with PBMC from distinct healthy donors. The efficiency of cytokine treatment was attested by the induction of HLA-DR expression on MSCs (Figures 1(a) and 2(a)). In order to evaluate the influence of osteodifferentiation process on the immunogenicity of MSCs, similar functional assays were performed using BM-derived and AT-derived MSCs committed to preosteoblastic MSCs as stimulating cells. The osteodifferentiation process was validated through the upregulation of ALP expression in osteodifferentiated MSCs (Figures 3(a), 3(b), 4(a), and 4(b)). Results show that both BM-derived and AT-derived MSCs committed to osteodifferentiation are still hypoimmunogenic whether they are pretreated or not with IFN-γ and TNF-α (Figures 3(c) and 4(c) and Tables 1 and 2). Then, we looked at whether combination of biomaterial (i.e., MBCP) with MSCs alters their immunogenicity. No differences were found between standard 2D-coculture conditions (MSC + PBMC) and 3D-coculture conditions (MSC + MBCP + PBMC). One representative allogenic combination is shown (Figure 5) for which the mean percentage of T cell alloproliferation is presented in Table 3 (n = 3 healthy donors). It is of note that the addition of MBCP to BM-derived MSCs treated or not with cytokines modifies slightly their immunogenicity, although no statistical difference was observed between both conditions (P > 0.1) (Table 3). Consequently, we can conclude that both the osteodifferentiation process and the presence of biomaterial (MBCP) did not abrogate the hypoimmunogenicity of MSCs.


Osteodifferentiated mesenchymal stem cells from bone marrow and adipose tissue express HLA-G and display immunomodulatory properties in HLA-mismatched settings: implications in bone repair therapy.

Montespan F, Deschaseaux F, Sensébé L, Carosella ED, Rouas-Freiss N - J Immunol Res (2014)

BM-derived MSCs committed to osteodifferentiation process express HLA-G and are hypoimmunogenic. (a and b) Expression of ALP and HLA-G5 was evaluated by intracellular flow cytometry analysis on BM-derived MSCs committed to 14-day osteodifferentiation process and pretreated with IFNγ and TNFα (Os-MSC-BM IFNγ/TNFα) or not (Os-MSC-BM). IgG1 was used as isotype control Ab. Tubulin was used as positive control of cell permeabilization. (c) PBMC from healthy individual (#00) were used as responder cells towards BM-derived MSCs committed to osteodifferentiation process and pretreated with IFNγ and TNFα (Os-MSC-BM IFNγ/TNFα) or not (Os-MSC-BM) as stimulating cells at various responder: stimulator ratios. Irradiated LCL* were used as positive control of T cell alloproliferation. Results are given as mean cpm ± s.e.m.; one representative experiment is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
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fig3: BM-derived MSCs committed to osteodifferentiation process express HLA-G and are hypoimmunogenic. (a and b) Expression of ALP and HLA-G5 was evaluated by intracellular flow cytometry analysis on BM-derived MSCs committed to 14-day osteodifferentiation process and pretreated with IFNγ and TNFα (Os-MSC-BM IFNγ/TNFα) or not (Os-MSC-BM). IgG1 was used as isotype control Ab. Tubulin was used as positive control of cell permeabilization. (c) PBMC from healthy individual (#00) were used as responder cells towards BM-derived MSCs committed to osteodifferentiation process and pretreated with IFNγ and TNFα (Os-MSC-BM IFNγ/TNFα) or not (Os-MSC-BM) as stimulating cells at various responder: stimulator ratios. Irradiated LCL* were used as positive control of T cell alloproliferation. Results are given as mean cpm ± s.e.m.; one representative experiment is shown.
Mentions: No PBMC alloproliferation was observed in response to various doses of allogenic MSCs derived from bone marrow (Figure 1(c)) or adipose tissue (Figure 2(c)) even after licensing with IFN-γ and TNF-α (Figures 1(c) and 2(c)). Tables 1 and 2 summarize the results obtained with PBMC from distinct healthy donors. The efficiency of cytokine treatment was attested by the induction of HLA-DR expression on MSCs (Figures 1(a) and 2(a)). In order to evaluate the influence of osteodifferentiation process on the immunogenicity of MSCs, similar functional assays were performed using BM-derived and AT-derived MSCs committed to preosteoblastic MSCs as stimulating cells. The osteodifferentiation process was validated through the upregulation of ALP expression in osteodifferentiated MSCs (Figures 3(a), 3(b), 4(a), and 4(b)). Results show that both BM-derived and AT-derived MSCs committed to osteodifferentiation are still hypoimmunogenic whether they are pretreated or not with IFN-γ and TNF-α (Figures 3(c) and 4(c) and Tables 1 and 2). Then, we looked at whether combination of biomaterial (i.e., MBCP) with MSCs alters their immunogenicity. No differences were found between standard 2D-coculture conditions (MSC + PBMC) and 3D-coculture conditions (MSC + MBCP + PBMC). One representative allogenic combination is shown (Figure 5) for which the mean percentage of T cell alloproliferation is presented in Table 3 (n = 3 healthy donors). It is of note that the addition of MBCP to BM-derived MSCs treated or not with cytokines modifies slightly their immunogenicity, although no statistical difference was observed between both conditions (P > 0.1) (Table 3). Consequently, we can conclude that both the osteodifferentiation process and the presence of biomaterial (MBCP) did not abrogate the hypoimmunogenicity of MSCs.

Bottom Line: Both MSCs and osteodifferentiated MSCs are hypoimmunogenic and exert immunomodulatory properties in HLA-mismatched settings as they suppress T cell alloproliferation in mixed lymphocyte reactions.Finally, addition of biomaterials that stimulate bone tissue formation did not modify MSC immune properties.As MSCs combine both abilities of osteoregeneration and immunomodulation, they may be considered as allogenic sources for the treatment of bone defects.

View Article: PubMed Central - PubMed

Affiliation: CEA, Institut des Maladies Emergentes et des Therapies Innovantes (IMETI), Service de Recherche en Hemato-Immunologie (SRHI), Hopital Saint-Louis, IUH, 1 avenue Claude Vellefaux, 75010 Paris, France ; Universite Paris Diderot, Sorbonne Paris Cité, IUH, Hopital Saint-Louis, UMR_E5, IUH, 1, avenue Claude Vellefaux, 75010 Paris, France.

ABSTRACT
Mesenchymal stem cells (MSCs) are multipotent cells that can be obtained from several sources such as bone marrow and adipose tissue. Depending on the culture conditions, they can differentiate into osteoblasts, chondroblasts, adipocytes, or neurons. In this regard, they constitute promising candidates for cell-based therapy aimed at repairing damaged tissues. In addition, MSCs display immunomodulatory properties through the expression of soluble factors including HLA-G. We here analyse both immunogenicity and immunosuppressive capacity of MSCs derived from bone marrow and adipose tissue before and after osteodifferentiation. Results show that HLA-G expression is maintained after osteodifferentiation and can be boosted in inflammatory conditions mimicked by the addition of IFN-γ and TNF-α. Both MSCs and osteodifferentiated MSCs are hypoimmunogenic and exert immunomodulatory properties in HLA-mismatched settings as they suppress T cell alloproliferation in mixed lymphocyte reactions. Finally, addition of biomaterials that stimulate bone tissue formation did not modify MSC immune properties. As MSCs combine both abilities of osteoregeneration and immunomodulation, they may be considered as allogenic sources for the treatment of bone defects.

Show MeSH