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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

Infiltrate of lymphocytes in salivary glands of NOD mice.A–E: H&E staining showing size of the lymphocytic infiltrations (delineated with a yellow line and arrows) in NOD that were untreated (A), CFA-treated (B), MSC+CFA (C), MSC cells (D). In 2 of the 5 NOD mice transplanted with MSC only, no lymphocytic infiltrates were noted (E). Scale bar: 140 um for all images. F: Graph showing distribution of focus score (n = 5 to 9 mice per group). The mean focus score was significantly higher in control (3.24) versus CFA (1.81), MSC (1.23), and MSC+CFA (1.8) NOD groups (* P = 0.02).
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pone-0038615-g002: Infiltrate of lymphocytes in salivary glands of NOD mice.A–E: H&E staining showing size of the lymphocytic infiltrations (delineated with a yellow line and arrows) in NOD that were untreated (A), CFA-treated (B), MSC+CFA (C), MSC cells (D). In 2 of the 5 NOD mice transplanted with MSC only, no lymphocytic infiltrates were noted (E). Scale bar: 140 um for all images. F: Graph showing distribution of focus score (n = 5 to 9 mice per group). The mean focus score was significantly higher in control (3.24) versus CFA (1.81), MSC (1.23), and MSC+CFA (1.8) NOD groups (* P = 0.02).

Mentions: To explore the mechanism underlying the preservation of SFR in NOD mice, salivary tissues were histologically analyzed for inflammatory signs, gene expression levels and cell chimerism. The focus scores (number of lymphocytic infiltrates in salivary tissue; Figure 2) of the CFA (1.81), MSC+CFA (1.8), and MSC (1.23) groups were all lower than the focus score of control NOD mice (3.24; P<0.05). The size of the lymphocytic infiltrate was smaller in NOD mice that received MSCs with or without CFA, when compared to untreated or CFA-treated mice (Figure 2 A, B, C, D, E). Interestingly, no lymphocytic infiltrates were detected in two out of five NOD mice transplanted with MSCs only (Figure 2E).


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)

Infiltrate of lymphocytes in salivary glands of NOD mice.A–E: H&E staining showing size of the lymphocytic infiltrations (delineated with a yellow line and arrows) in NOD that were untreated (A), CFA-treated (B), MSC+CFA (C), MSC cells (D). In 2 of the 5 NOD mice transplanted with MSC only, no lymphocytic infiltrates were noted (E). Scale bar: 140 um for all images. F: Graph showing distribution of focus score (n = 5 to 9 mice per group). The mean focus score was significantly higher in control (3.24) versus CFA (1.81), MSC (1.23), and MSC+CFA (1.8) NOD groups (* P = 0.02).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038615-g002: Infiltrate of lymphocytes in salivary glands of NOD mice.A–E: H&E staining showing size of the lymphocytic infiltrations (delineated with a yellow line and arrows) in NOD that were untreated (A), CFA-treated (B), MSC+CFA (C), MSC cells (D). In 2 of the 5 NOD mice transplanted with MSC only, no lymphocytic infiltrates were noted (E). Scale bar: 140 um for all images. F: Graph showing distribution of focus score (n = 5 to 9 mice per group). The mean focus score was significantly higher in control (3.24) versus CFA (1.81), MSC (1.23), and MSC+CFA (1.8) NOD groups (* P = 0.02).
Mentions: To explore the mechanism underlying the preservation of SFR in NOD mice, salivary tissues were histologically analyzed for inflammatory signs, gene expression levels and cell chimerism. The focus scores (number of lymphocytic infiltrates in salivary tissue; Figure 2) of the CFA (1.81), MSC+CFA (1.8), and MSC (1.23) groups were all lower than the focus score of control NOD mice (3.24; P<0.05). The size of the lymphocytic infiltrate was smaller in NOD mice that received MSCs with or without CFA, when compared to untreated or CFA-treated mice (Figure 2 A, B, C, D, E). Interestingly, no lymphocytic infiltrates were detected in two out of five NOD mice transplanted with MSCs only (Figure 2E).

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