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Mechanisms of the immunosuppressive effects of mouse adipose tissue-derived mesenchymal stromal cells on mouse alloreactively stimulated spleen cells.

Nagaya R, Mizuno-Kamiya M, Takayama E, Kawaki H, Onoe I, Tanabe T, Nagahara K, Kondoh N - Exp Ther Med (2013)

Bottom Line: The suppressive effect of ASCs mediated by direct cell contact was partially reversed following knockdown of β2 microglobulin, a component of the major histocompatibility complex (MHC) class I molecule, with siRNA.The results of the study demonstrated that ASCs have significant immune modulatory effects on alloreactively stimulated spleen cells.The effects of ASCs on spleen cells are dependent on soluble factor(s) and cell contact, which is mediated by the MHC class I complex on ASCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Implantology, Asahi University School of Dentistry, Mizuho-shi, Gifu 501-0296, Japan.

ABSTRACT
The mechanisms of immunomodulation by mesenchymal stromal cells remain poorly understood. In this study, the effects of mouse adipose tissue-derived mesenchymal stromal cells (ASCs) on mouse spleen cells alloreactively stimulated by anti-CD3 and anti-CD28 antibody-coated (anti-CD3/CD28) beads were observed. Production of interferon-γ by the anti-CD3/CD28 bead-stimulated spleen cells was significantly suppressed in co-culture with ASCs. However, an augmented intensity of CD69 on the stimulated spleen cells was not suppressed in the presence of ASCs. The immunosuppressive effects of ASCs were partially mediated by one or more soluble factors (26% suppression). However, the ASCs require cell-cell contact in order to maximally exert suppression (88%). The suppressive effect of ASCs mediated by direct cell contact was partially reversed following knockdown of β2 microglobulin, a component of the major histocompatibility complex (MHC) class I molecule, with siRNA. The results of the study demonstrated that ASCs have significant immune modulatory effects on alloreactively stimulated spleen cells. The effects of ASCs on spleen cells are dependent on soluble factor(s) and cell contact, which is mediated by the MHC class I complex on ASCs.

No MeSH data available.


Related in: MedlinePlus

Flow cytometric analysis of T-cell subsets in CD45+ spleen cells. Spleen cells (4×105) were cultured in the presence or absence of 4×105 anti-CD3/CD28 beads and ASCs for 12 h. The cells were harvested, then FACS analyses were performed using anti-CD45 (PerCP-Cy™5.5), anti-CD4 (PE), anti-CD8 (FITC) and anti-CD69 (APC) antibodies. The percentage of each cell subpopulation (denoted on the right side) among the CD45+ total lymphocytes are indicated. ASCs, adipose tissue-derived mesenchymal stromal cells.
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f3-etm-07-01-0017: Flow cytometric analysis of T-cell subsets in CD45+ spleen cells. Spleen cells (4×105) were cultured in the presence or absence of 4×105 anti-CD3/CD28 beads and ASCs for 12 h. The cells were harvested, then FACS analyses were performed using anti-CD45 (PerCP-Cy™5.5), anti-CD4 (PE), anti-CD8 (FITC) and anti-CD69 (APC) antibodies. The percentage of each cell subpopulation (denoted on the right side) among the CD45+ total lymphocytes are indicated. ASCs, adipose tissue-derived mesenchymal stromal cells.

Mentions: To investigate whether the activation status of T cells is changed by ASCs or not, the expression of CD69 on stimulated spleen cells was analyzed by flow cytometry (Fig. 3). CD69 is the earliest inducible cell surface glycoprotein acquired during lymphoid activation, which is involved in lymphocyte proliferation and function (27). In order to select all leukocytes, CD45+ cells were first gated (100%). CD69 positive/negative fractions among CD4+/CD8− and CD4−/CD8+ T subsets were compared. The results are summarized in Fig. 3. No CD69+ cells were detected in CD45+ un-stimulated spleen cells. Following stimulation with the anti-CD3/CD28 beads, CD69+ populations emerged in the CD4+/CD8− and CD4−/CD8+ T subsets, as 7.6% and 5.5%, respectively. In the presence of ASCs, the production of IFN-γ was greatly suppressed; however, certain populations of CD69+ continued to exist in the CD4+/CD8− and CD4−/CD8+ subsets, as 9.1% and 5.4%, respectively.


Mechanisms of the immunosuppressive effects of mouse adipose tissue-derived mesenchymal stromal cells on mouse alloreactively stimulated spleen cells.

Nagaya R, Mizuno-Kamiya M, Takayama E, Kawaki H, Onoe I, Tanabe T, Nagahara K, Kondoh N - Exp Ther Med (2013)

Flow cytometric analysis of T-cell subsets in CD45+ spleen cells. Spleen cells (4×105) were cultured in the presence or absence of 4×105 anti-CD3/CD28 beads and ASCs for 12 h. The cells were harvested, then FACS analyses were performed using anti-CD45 (PerCP-Cy™5.5), anti-CD4 (PE), anti-CD8 (FITC) and anti-CD69 (APC) antibodies. The percentage of each cell subpopulation (denoted on the right side) among the CD45+ total lymphocytes are indicated. ASCs, adipose tissue-derived mesenchymal stromal cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3860983&req=5

f3-etm-07-01-0017: Flow cytometric analysis of T-cell subsets in CD45+ spleen cells. Spleen cells (4×105) were cultured in the presence or absence of 4×105 anti-CD3/CD28 beads and ASCs for 12 h. The cells were harvested, then FACS analyses were performed using anti-CD45 (PerCP-Cy™5.5), anti-CD4 (PE), anti-CD8 (FITC) and anti-CD69 (APC) antibodies. The percentage of each cell subpopulation (denoted on the right side) among the CD45+ total lymphocytes are indicated. ASCs, adipose tissue-derived mesenchymal stromal cells.
Mentions: To investigate whether the activation status of T cells is changed by ASCs or not, the expression of CD69 on stimulated spleen cells was analyzed by flow cytometry (Fig. 3). CD69 is the earliest inducible cell surface glycoprotein acquired during lymphoid activation, which is involved in lymphocyte proliferation and function (27). In order to select all leukocytes, CD45+ cells were first gated (100%). CD69 positive/negative fractions among CD4+/CD8− and CD4−/CD8+ T subsets were compared. The results are summarized in Fig. 3. No CD69+ cells were detected in CD45+ un-stimulated spleen cells. Following stimulation with the anti-CD3/CD28 beads, CD69+ populations emerged in the CD4+/CD8− and CD4−/CD8+ T subsets, as 7.6% and 5.5%, respectively. In the presence of ASCs, the production of IFN-γ was greatly suppressed; however, certain populations of CD69+ continued to exist in the CD4+/CD8− and CD4−/CD8+ subsets, as 9.1% and 5.4%, respectively.

Bottom Line: The suppressive effect of ASCs mediated by direct cell contact was partially reversed following knockdown of β2 microglobulin, a component of the major histocompatibility complex (MHC) class I molecule, with siRNA.The results of the study demonstrated that ASCs have significant immune modulatory effects on alloreactively stimulated spleen cells.The effects of ASCs on spleen cells are dependent on soluble factor(s) and cell contact, which is mediated by the MHC class I complex on ASCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Implantology, Asahi University School of Dentistry, Mizuho-shi, Gifu 501-0296, Japan.

ABSTRACT
The mechanisms of immunomodulation by mesenchymal stromal cells remain poorly understood. In this study, the effects of mouse adipose tissue-derived mesenchymal stromal cells (ASCs) on mouse spleen cells alloreactively stimulated by anti-CD3 and anti-CD28 antibody-coated (anti-CD3/CD28) beads were observed. Production of interferon-γ by the anti-CD3/CD28 bead-stimulated spleen cells was significantly suppressed in co-culture with ASCs. However, an augmented intensity of CD69 on the stimulated spleen cells was not suppressed in the presence of ASCs. The immunosuppressive effects of ASCs were partially mediated by one or more soluble factors (26% suppression). However, the ASCs require cell-cell contact in order to maximally exert suppression (88%). The suppressive effect of ASCs mediated by direct cell contact was partially reversed following knockdown of β2 microglobulin, a component of the major histocompatibility complex (MHC) class I molecule, with siRNA. The results of the study demonstrated that ASCs have significant immune modulatory effects on alloreactively stimulated spleen cells. The effects of ASCs on spleen cells are dependent on soluble factor(s) and cell contact, which is mediated by the MHC class I complex on ASCs.

No MeSH data available.


Related in: MedlinePlus