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Mesenchymal stromal cells improve salivary function and reduce lymphocytic infiltrates in mice with Sjögren's-like disease.

Khalili S, Liu Y, Kornete M, Roescher N, Kodama S, Peterson A, Piccirillo CA, Tran SD - PLoS ONE (2012)

Bottom Line: Both MSC and MSC+CFA groups prevented loss of saliva flow and reduced lymphocytic infiltrations in SGs.MSC-therapy alone reduced inflammation (TNF-α, TGF-β), but the combination of MSC+CFA reduced inflammation and increased the regenerative potential of SGs (FGF-2, EGF).The combined use of MSC+CFA was effective in both preventing saliva secretion loss and reducing lymphocytic influx in salivary glands.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Dentistry, McGill University, Montreal, Quebec, Canada.

ABSTRACT

Background: Non-obese diabetic (NOD) mice develop Sjögren's-like disease (SS-like) with loss of saliva flow and increased lymphocytic infiltrates in salivary glands (SGs). There are recent reports using multipotent mesenchymal stromal cells (MSCs) as a therapeutic strategy for autoimmune diseases due to their anti-inflammatory and immunomodulatory capabilities. This paper proposed a combined immuno- and cell-based therapy consisting of: A) an injection of complete Freund's adjuvant (CFA) to eradicate autoreactive T lymphocytes, and B) transplantations of MSCs to reselect lymphocytes. The objective of this was to test the effectiveness of CD45(-)/TER119(-) cells (MSCs) in re-establishing salivary function and in reducing the number of lymphocytic infiltrates (foci) in SGs. The second objective was to study if the mechanisms underlying a decrease in inflammation (focus score) was due to CFA, MSCs, or CFA+MSCs combined.

Methodology/principal findings: Donor MSCs were isolated from bones of male transgenic eGFP mice. Eight week-old female NOD mice received one of the following treatments: insulin, CFA, MSC, or CFA+MSC (combined therapy). Mice were followed for 14 weeks post-therapy. CD45(-)/TER119(-) cells demonstrated characteristics of MSCs as they were positive for Sca-1, CD106, CD105, CD73, CD29, CD44, negative for CD45, TER119, CD11b, had high number of CFU-F, and differentiated into osteocytes, chondrocytes and adipocytes. Both MSC and MSC+CFA groups prevented loss of saliva flow and reduced lymphocytic infiltrations in SGs. Moreover, the influx of T and B cells decreased in all foci in MSC and MSC+CFA groups, while the frequency of Foxp3(+) (T(reg)) cell was increased. MSC-therapy alone reduced inflammation (TNF-α, TGF-β), but the combination of MSC+CFA reduced inflammation and increased the regenerative potential of SGs (FGF-2, EGF).

Conclusions/significance: The combined use of MSC+CFA was effective in both preventing saliva secretion loss and reducing lymphocytic influx in salivary glands.

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Related in: MedlinePlus

Immunohistochemical staining of salivary glands at week 22 (n = 3 to 5 mice per group).Cells positive for Foxp3+ (regulatory T cells), CD8+ (T cytotoxic), CD4+ (T helper), BAFF and CD19+ (B cells) appeared brown in the foci. Graphs on the right represent percentage of positive cells occupying the area of the focus score. Regulatory T cells (Foxp3+) increased in MSC and MSC+CFA groups (P<0.05) while T cytotoxic (CD8+), T helper (CD4+), BAFF, and B cell (CD19+) decreased after MSC or MSC+CFA treatment, when compared to control and CFA groups (* P<0.05).
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pone-0038615-g005: Immunohistochemical staining of salivary glands at week 22 (n = 3 to 5 mice per group).Cells positive for Foxp3+ (regulatory T cells), CD8+ (T cytotoxic), CD4+ (T helper), BAFF and CD19+ (B cells) appeared brown in the foci. Graphs on the right represent percentage of positive cells occupying the area of the focus score. Regulatory T cells (Foxp3+) increased in MSC and MSC+CFA groups (P<0.05) while T cytotoxic (CD8+), T helper (CD4+), BAFF, and B cell (CD19+) decreased after MSC or MSC+CFA treatment, when compared to control and CFA groups (* P<0.05).

Mentions: Proportions of immune cells in focus score, such as T helper (CD4+), T cytotoxic (CD8+), B cell (CD19+), and BAFF, was fewer in MSC and MSC+CFA groups when compared to control or CFA groups (Figure 5; P<0.05). However, regulatory T cells (Treg Foxp3+) cells increased in MSC and MSC+CFA groups (P<0.05). No significant difference was found for natural killer (CD49b+) and plasmacytoid dendritic cells (CD11c+).


Mesenchymal stromal cells improve salivary function and reduce lymphocytic infiltrates in mice with Sjögren's-like disease.

Khalili S, Liu Y, Kornete M, Roescher N, Kodama S, Peterson A, Piccirillo CA, Tran SD - PLoS ONE (2012)

Immunohistochemical staining of salivary glands at week 22 (n = 3 to 5 mice per group).Cells positive for Foxp3+ (regulatory T cells), CD8+ (T cytotoxic), CD4+ (T helper), BAFF and CD19+ (B cells) appeared brown in the foci. Graphs on the right represent percentage of positive cells occupying the area of the focus score. Regulatory T cells (Foxp3+) increased in MSC and MSC+CFA groups (P<0.05) while T cytotoxic (CD8+), T helper (CD4+), BAFF, and B cell (CD19+) decreased after MSC or MSC+CFA treatment, when compared to control and CFA groups (* P<0.05).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3369846&req=5

pone-0038615-g005: Immunohistochemical staining of salivary glands at week 22 (n = 3 to 5 mice per group).Cells positive for Foxp3+ (regulatory T cells), CD8+ (T cytotoxic), CD4+ (T helper), BAFF and CD19+ (B cells) appeared brown in the foci. Graphs on the right represent percentage of positive cells occupying the area of the focus score. Regulatory T cells (Foxp3+) increased in MSC and MSC+CFA groups (P<0.05) while T cytotoxic (CD8+), T helper (CD4+), BAFF, and B cell (CD19+) decreased after MSC or MSC+CFA treatment, when compared to control and CFA groups (* P<0.05).
Mentions: Proportions of immune cells in focus score, such as T helper (CD4+), T cytotoxic (CD8+), B cell (CD19+), and BAFF, was fewer in MSC and MSC+CFA groups when compared to control or CFA groups (Figure 5; P<0.05). However, regulatory T cells (Treg Foxp3+) cells increased in MSC and MSC+CFA groups (P<0.05). No significant difference was found for natural killer (CD49b+) and plasmacytoid dendritic cells (CD11c+).

Bottom Line: Both MSC and MSC+CFA groups prevented loss of saliva flow and reduced lymphocytic infiltrations in SGs.MSC-therapy alone reduced inflammation (TNF-α, TGF-β), but the combination of MSC+CFA reduced inflammation and increased the regenerative potential of SGs (FGF-2, EGF).The combined use of MSC+CFA was effective in both preventing saliva secretion loss and reducing lymphocytic influx in salivary glands.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Dentistry, McGill University, Montreal, Quebec, Canada.

ABSTRACT

Background: Non-obese diabetic (NOD) mice develop Sjögren's-like disease (SS-like) with loss of saliva flow and increased lymphocytic infiltrates in salivary glands (SGs). There are recent reports using multipotent mesenchymal stromal cells (MSCs) as a therapeutic strategy for autoimmune diseases due to their anti-inflammatory and immunomodulatory capabilities. This paper proposed a combined immuno- and cell-based therapy consisting of: A) an injection of complete Freund's adjuvant (CFA) to eradicate autoreactive T lymphocytes, and B) transplantations of MSCs to reselect lymphocytes. The objective of this was to test the effectiveness of CD45(-)/TER119(-) cells (MSCs) in re-establishing salivary function and in reducing the number of lymphocytic infiltrates (foci) in SGs. The second objective was to study if the mechanisms underlying a decrease in inflammation (focus score) was due to CFA, MSCs, or CFA+MSCs combined.

Methodology/principal findings: Donor MSCs were isolated from bones of male transgenic eGFP mice. Eight week-old female NOD mice received one of the following treatments: insulin, CFA, MSC, or CFA+MSC (combined therapy). Mice were followed for 14 weeks post-therapy. CD45(-)/TER119(-) cells demonstrated characteristics of MSCs as they were positive for Sca-1, CD106, CD105, CD73, CD29, CD44, negative for CD45, TER119, CD11b, had high number of CFU-F, and differentiated into osteocytes, chondrocytes and adipocytes. Both MSC and MSC+CFA groups prevented loss of saliva flow and reduced lymphocytic infiltrations in SGs. Moreover, the influx of T and B cells decreased in all foci in MSC and MSC+CFA groups, while the frequency of Foxp3(+) (T(reg)) cell was increased. MSC-therapy alone reduced inflammation (TNF-α, TGF-β), but the combination of MSC+CFA reduced inflammation and increased the regenerative potential of SGs (FGF-2, EGF).

Conclusions/significance: The combined use of MSC+CFA was effective in both preventing saliva secretion loss and reducing lymphocytic influx in salivary glands.

Show MeSH
Related in: MedlinePlus