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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

Absence of eGFP and male MSC (CD45−/TER119−) donor cells in salivary tissues of female NOD mice.Top panel: eGFP detection of MSC donor cells by immunostaining. (A) Salivary tissues from the eGFP donor mouse (positive control) versus (B) the non-eGFP mouse (negative control). (C) NOD mice transplanted with MSCs were negative for eGFP cells. Middle panel: Y-chromosome detection of male MSCs by FISH. (D) male and (E) female salivary tissues used as positive and negative controls, respectively. (F) Female NOD mice transplanted with male MSCs showed no Y-chromosome signal in their salivary glands. Bottom panel: (G) CByB6F1-eGFP transgenic mouse (arrow) and (H) the isolated donor eGFP MSCs before transplantation. Cell nuclei are stained in blue (Hoechst 33258). (I) PCR amplification did not detect the Y-chromosome and eGFP signal in salivary glands of MSCs transplanted NOD mice (n = 11).
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pone-0038615-g008: Absence of eGFP and male MSC (CD45−/TER119−) donor cells in salivary tissues of female NOD mice.Top panel: eGFP detection of MSC donor cells by immunostaining. (A) Salivary tissues from the eGFP donor mouse (positive control) versus (B) the non-eGFP mouse (negative control). (C) NOD mice transplanted with MSCs were negative for eGFP cells. Middle panel: Y-chromosome detection of male MSCs by FISH. (D) male and (E) female salivary tissues used as positive and negative controls, respectively. (F) Female NOD mice transplanted with male MSCs showed no Y-chromosome signal in their salivary glands. Bottom panel: (G) CByB6F1-eGFP transgenic mouse (arrow) and (H) the isolated donor eGFP MSCs before transplantation. Cell nuclei are stained in blue (Hoechst 33258). (I) PCR amplification did not detect the Y-chromosome and eGFP signal in salivary glands of MSCs transplanted NOD mice (n = 11).

Mentions: Fluorescence in situ hybridization (FISH) and immunostaining were used to detect the male (Y-chromosome) and eGFP markers of transplanted MSCs in female NOD salivary tissues, respectively. No Y-chromosome or eGFP-positive cells were detected (Figure 8). Salivary tissues also were investigated by PCR for the presence of the Y-chromosome and eGFP; no signal was detected for either (Figure 8).


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)

Absence of eGFP and male MSC (CD45−/TER119−) donor cells in salivary tissues of female NOD mice.Top panel: eGFP detection of MSC donor cells by immunostaining. (A) Salivary tissues from the eGFP donor mouse (positive control) versus (B) the non-eGFP mouse (negative control). (C) NOD mice transplanted with MSCs were negative for eGFP cells. Middle panel: Y-chromosome detection of male MSCs by FISH. (D) male and (E) female salivary tissues used as positive and negative controls, respectively. (F) Female NOD mice transplanted with male MSCs showed no Y-chromosome signal in their salivary glands. Bottom panel: (G) CByB6F1-eGFP transgenic mouse (arrow) and (H) the isolated donor eGFP MSCs before transplantation. Cell nuclei are stained in blue (Hoechst 33258). (I) PCR amplification did not detect the Y-chromosome and eGFP signal in salivary glands of MSCs transplanted NOD mice (n = 11).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038615-g008: Absence of eGFP and male MSC (CD45−/TER119−) donor cells in salivary tissues of female NOD mice.Top panel: eGFP detection of MSC donor cells by immunostaining. (A) Salivary tissues from the eGFP donor mouse (positive control) versus (B) the non-eGFP mouse (negative control). (C) NOD mice transplanted with MSCs were negative for eGFP cells. Middle panel: Y-chromosome detection of male MSCs by FISH. (D) male and (E) female salivary tissues used as positive and negative controls, respectively. (F) Female NOD mice transplanted with male MSCs showed no Y-chromosome signal in their salivary glands. Bottom panel: (G) CByB6F1-eGFP transgenic mouse (arrow) and (H) the isolated donor eGFP MSCs before transplantation. Cell nuclei are stained in blue (Hoechst 33258). (I) PCR amplification did not detect the Y-chromosome and eGFP signal in salivary glands of MSCs transplanted NOD mice (n = 11).
Mentions: Fluorescence in situ hybridization (FISH) and immunostaining were used to detect the male (Y-chromosome) and eGFP markers of transplanted MSCs in female NOD salivary tissues, respectively. No Y-chromosome or eGFP-positive cells were detected (Figure 8). Salivary tissues also were investigated by PCR for the presence of the Y-chromosome and eGFP; no signal was detected for either (Figure 8).

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