Limits...
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.

Show MeSH

Related in: MedlinePlus

Salivary flow rates (SFRs) of NOD mice.SFRs in MSC+CFA (black circle; n = 10) and MSC (black square; n = 5) groups did not decrease during the follow-up period (22 wk of age) and were significantly higher than SFRs of CFA-treated or control NOD groups (n = 5 per group; P<0.05). SFRs in CFA (triangular) or control (untreated; open circle) groups continued to decrease during the follow-up period (* P<0.05).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3369846&req=5

pone-0038615-g001: Salivary flow rates (SFRs) of NOD mice.SFRs in MSC+CFA (black circle; n = 10) and MSC (black square; n = 5) groups did not decrease during the follow-up period (22 wk of age) and were significantly higher than SFRs of CFA-treated or control NOD groups (n = 5 per group; P<0.05). SFRs in CFA (triangular) or control (untreated; open circle) groups continued to decrease during the follow-up period (* P<0.05).

Mentions: Salivary gland function was assessed by measuring the salivary flow rate (SFR) of 8-week old NOD mice at baseline and for 14 weeks post-therapy. SFR directly reflects function of the glands and its decrease is the major clinical finding in patients with SS. In untreated NOD mice, SFR continued to deteriorate during the 14 weeks follow-up period (Figure 1). SFR of CFA-treated NOD mice (with no cell transplants) also deteriorated but to a lesser extent. SFRs of NOD mice that received the combined immuno- and cell-based therapy (CD45−/TER119− MSCs with CFA) or the cell-based therapy alone (CD45−/TER119− MSCs) were maintained during the 14 weeks post-therapy period and were statistically higher than SFRs of untreated and CFA-treated NOD mice (P<0.05; Figure 1). The composition and quality of saliva was examined for EGF, amylase, and total protein concentrations. These levels did not change significantly between pre-therapy (8-week NOD mice, prior to SS-like) and post-therapy (Figure S1). Blood glucose levels were monitored as NOD mice are known to develop type I diabetes. Sixty percent of control NOD mice (only receiving injections of insulin) developed diabetes at 22 weeks of age (Figure S2; P<0.05). However, 80%–90% NOD mice in the 3 other treated groups (CFA, MSC, and MSC+CFA) showed a relatively stable blood sugar level (normoglycemia) until 22 weeks of age.


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)

Salivary flow rates (SFRs) of NOD mice.SFRs in MSC+CFA (black circle; n = 10) and MSC (black square; n = 5) groups did not decrease during the follow-up period (22 wk of age) and were significantly higher than SFRs of CFA-treated or control NOD groups (n = 5 per group; P<0.05). SFRs in CFA (triangular) or control (untreated; open circle) groups continued to decrease during the follow-up period (* P<0.05).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038615-g001: Salivary flow rates (SFRs) of NOD mice.SFRs in MSC+CFA (black circle; n = 10) and MSC (black square; n = 5) groups did not decrease during the follow-up period (22 wk of age) and were significantly higher than SFRs of CFA-treated or control NOD groups (n = 5 per group; P<0.05). SFRs in CFA (triangular) or control (untreated; open circle) groups continued to decrease during the follow-up period (* P<0.05).
Mentions: Salivary gland function was assessed by measuring the salivary flow rate (SFR) of 8-week old NOD mice at baseline and for 14 weeks post-therapy. SFR directly reflects function of the glands and its decrease is the major clinical finding in patients with SS. In untreated NOD mice, SFR continued to deteriorate during the 14 weeks follow-up period (Figure 1). SFR of CFA-treated NOD mice (with no cell transplants) also deteriorated but to a lesser extent. SFRs of NOD mice that received the combined immuno- and cell-based therapy (CD45−/TER119− MSCs with CFA) or the cell-based therapy alone (CD45−/TER119− MSCs) were maintained during the 14 weeks post-therapy period and were statistically higher than SFRs of untreated and CFA-treated NOD mice (P<0.05; Figure 1). The composition and quality of saliva was examined for EGF, amylase, and total protein concentrations. These levels did not change significantly between pre-therapy (8-week NOD mice, prior to SS-like) and post-therapy (Figure S1). Blood glucose levels were monitored as NOD mice are known to develop type I diabetes. Sixty percent of control NOD mice (only receiving injections of insulin) developed diabetes at 22 weeks of age (Figure S2; P<0.05). However, 80%–90% NOD mice in the 3 other treated groups (CFA, MSC, and MSC+CFA) showed a relatively stable blood sugar level (normoglycemia) until 22 weeks of age.

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