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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

Measurements of TNFα and EGF concentrations in saliva and serum.Upper panel (A), as expected, the level of TNFα in saliva was significantly higher (*P<0.05) in non-treated NOD mice at week 22 (established Sjögren's-like disease) when compared to week 8 (before disease onset). At the end of the experimentation period (week 22), treated MSC and MSC+CFA mice had two times less TNFα levels than non-treated NOD (*P<0.05). Lower panel (B), concentrations of serum EGF was significantly higher in MSC+CFA group versus control, CFA, and MSC groups (n = 4 to 8 mice per group; P<0.05).
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pone-0038615-g004: Measurements of TNFα and EGF concentrations in saliva and serum.Upper panel (A), as expected, the level of TNFα in saliva was significantly higher (*P<0.05) in non-treated NOD mice at week 22 (established Sjögren's-like disease) when compared to week 8 (before disease onset). At the end of the experimentation period (week 22), treated MSC and MSC+CFA mice had two times less TNFα levels than non-treated NOD (*P<0.05). Lower panel (B), concentrations of serum EGF was significantly higher in MSC+CFA group versus control, CFA, and MSC groups (n = 4 to 8 mice per group; P<0.05).

Mentions: Expression of several important growth factor/cytokine genes involved in the pathogenesis of SS (TNF-α, TGF-β1) and in salivary tissue regeneration/homeostasis (EGF, FGF2, IGF-IR, AQP5) was measured (Figure 3). TNF-α mRNA were ∼4 fold lower in CFA-treated and MSC+CFA versus control NOD mice (Figure 3 upper panel; P<0.05). NOD mice transplanted with only MSCs had the lowest expression of TNF-α (10 fold lower than control NOD and 5 fold lower than CFA or MSC+CFA; P<0.05). TNF-α concentration in saliva of MSC and MSC+CFA groups were twice lower than those of NOD controls (Figure 4 upper panel; P<0.05). Transforming growth factor β1 (TGF-β1) mRNA accumulated 2.5 fold lower in MSC and MSC+CFA mice versus control NOD (Figure 3; P<0.05). Epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF-2) were 3 to 4 fold upregulated in MSC group when compared to controls (Figure 3; P<0.05). NOD treated with the combined immuno- and cell-based therapy (MSC+CFA) showed the highest upregulation of EGF and FGF2 (10 and 5-fold, respectively). Insulin-like growth factor receptor I (IGF-IR) was upregulated (4–5 fold) in the MSC and MSC+CFA groups when compared to control NOD (P<0.05). The water channel protein, aquaporin-5 (AQP5), was ∼4 fold upregulated in treated animals (CFA alone, MSC with or without CFA) (P<0.05). The concentration of serum EGF was 10 times higher in the MSC+CFA treated group when compared to control NOD, and 5 times higher than the two other groups (only CFA, or only MSC) (Figure 4 lower panel; P<0.05).


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)

Measurements of TNFα and EGF concentrations in saliva and serum.Upper panel (A), as expected, the level of TNFα in saliva was significantly higher (*P<0.05) in non-treated NOD mice at week 22 (established Sjögren's-like disease) when compared to week 8 (before disease onset). At the end of the experimentation period (week 22), treated MSC and MSC+CFA mice had two times less TNFα levels than non-treated NOD (*P<0.05). Lower panel (B), concentrations of serum EGF was significantly higher in MSC+CFA group versus control, CFA, and MSC groups (n = 4 to 8 mice per group; P<0.05).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038615-g004: Measurements of TNFα and EGF concentrations in saliva and serum.Upper panel (A), as expected, the level of TNFα in saliva was significantly higher (*P<0.05) in non-treated NOD mice at week 22 (established Sjögren's-like disease) when compared to week 8 (before disease onset). At the end of the experimentation period (week 22), treated MSC and MSC+CFA mice had two times less TNFα levels than non-treated NOD (*P<0.05). Lower panel (B), concentrations of serum EGF was significantly higher in MSC+CFA group versus control, CFA, and MSC groups (n = 4 to 8 mice per group; P<0.05).
Mentions: Expression of several important growth factor/cytokine genes involved in the pathogenesis of SS (TNF-α, TGF-β1) and in salivary tissue regeneration/homeostasis (EGF, FGF2, IGF-IR, AQP5) was measured (Figure 3). TNF-α mRNA were ∼4 fold lower in CFA-treated and MSC+CFA versus control NOD mice (Figure 3 upper panel; P<0.05). NOD mice transplanted with only MSCs had the lowest expression of TNF-α (10 fold lower than control NOD and 5 fold lower than CFA or MSC+CFA; P<0.05). TNF-α concentration in saliva of MSC and MSC+CFA groups were twice lower than those of NOD controls (Figure 4 upper panel; P<0.05). Transforming growth factor β1 (TGF-β1) mRNA accumulated 2.5 fold lower in MSC and MSC+CFA mice versus control NOD (Figure 3; P<0.05). Epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF-2) were 3 to 4 fold upregulated in MSC group when compared to controls (Figure 3; P<0.05). NOD treated with the combined immuno- and cell-based therapy (MSC+CFA) showed the highest upregulation of EGF and FGF2 (10 and 5-fold, respectively). Insulin-like growth factor receptor I (IGF-IR) was upregulated (4–5 fold) in the MSC and MSC+CFA groups when compared to control NOD (P<0.05). The water channel protein, aquaporin-5 (AQP5), was ∼4 fold upregulated in treated animals (CFA alone, MSC with or without CFA) (P<0.05). The concentration of serum EGF was 10 times higher in the MSC+CFA treated group when compared to control NOD, and 5 times higher than the two other groups (only CFA, or only MSC) (Figure 4 lower panel; P<0.05).

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