Int J Stem Cells.  2017 Nov;10(2):160-168. 10.15283/ijsc17014.

Treatment with Allogenic Mesenchymal Stromal Cells in a Murine Model of Systemic Lupus Erythematosus

• Tani C
• Vagnani S
• Carli L
• Querci F
• Kühl AA
• Spieckermann S
• Cieluch CP
• Pacini S
• Fazzi R
• Mosca M

• KMID: 2400870
• DOI: http://doi.org/10.15283/ijsc17014

Abstract

OBJECTIVE
Pre-clinical and uncontrolled studies in patients with systemic lupus erythematosus (SLE) showed that mesenchymal stromal cells (MSCs) have a potential therapeutic role in refractory cases. The optimal therapeutic strategy in these patients remain to be elucidated. Our aim was to test the hypothesis that repeated administrations of 1×10⁶/kg body weight of allogenic MSCs, that is a significantly lower dosage with respect to the fixed 1×10⁶ MSC used in animal models, can be effective in improving the clinical course of a murine SLE model.
METHODS
Bone marrow derived MSCs were obtained from 12-week-old C57BL/6J mice. Seventy-five 8 weeks old female NZ mice were randomly assigned to receive via caudal vein the following alternative treatments: 1) single infusion of 10⁶ MSCs/kg body weight at 18 weeks of age (NZ(s18)) or at at 22 weeks of age (NZ(s22)); 2) multiple monthly infusions of 10⁶ MSCs/kg body weight starting at 18 weeks of age (NZ(M18)) or at 22 weeks of age (NZ(M22)); 3) saline infusions (NZ(c)) Fifteen 8 weeks old C57BL/6J mice (Envigo, Huntingdon, UK) were used as untreated controls (C). Weekly, body weight was recorded and twenty-four hour urines were collected by metabolic cages for each animal; proteinuria was detected by dipstick analysis. At sacrifice, peripheral blood samples were collected from mice and anti-dsDNA antibodies were detected by enzyme immunoassorbent assay (ELISA) method using commercial kits. At sacrifice, kidneys were analyzed for histopathology and immunohistochemical analysis for B220, CD4, MPO, CD4⁺Foxp3, F40/80 infiltration was performed.
RESULTS
Proteinuria occurrence was delayed NZ(S) and NZ(M) mice, no differences were observed in anti-dsDNA autoantibody titer among the groups at the different time-points; at 36 weeks, no significant differences were observed in term of nephritis scores. Inflammatory cells deposition (MPO and F4/80 positive cells) in NZM was significantly higher than in NZ and NZ(S). An overexpression of B lymphocytes (B220) was found in NZ(M) while T regulatory cells (CD4⁺ Foxp3⁺ cells) were reduced in both NZ(S) and NZ(M) with respect to NZ(c).
CONCLUSIONS
Overall, our study failed to show a positive effect of a treatment with murine MSCs in this model and, for some aspects, even deleterious results seem to be observed.

Keyword

Mesenchymal stromal cells; Systemic lupus erythematosus; Animal model; Lupus nephritis

MeSH Terms

Animals
Antibodies
B-Lymphocytes
Body Weight
Bone Marrow
Female
Humans
Kidney
Lupus Erythematosus, Systemic*
Lupus Nephritis
Mesenchymal Stromal Cells*
Methods
Mice
Models, Animal
Nephritis
Proteinuria
Veins
Antibodies

Figure

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  Fig. 1 24-hour proteinuria (mg/24 h).

• 

  Fig. 2 Anti-dsDNA levels at different time points. Results are expressed as optical density (OD) (mean of the optical density readings serum at 450 nm) as measured by a microtiter plate reader (Ultrospec2000, Pharmacia Biotech).

• 

  Fig. 3 Kidney Histology and Immunohistochemistry. (A) NZ HE×100 scale bar 100 μm, (B) NZs HE × 100 scale bar 100 μm, (C) NZm HE×100 scale bar 100 μm, (D) NZ CD3-AP B220×100 scale bar 100 μm, (E) NZs CD3-AP B220×100 scale bar 100 μm, (F) NZm CD3-AP B220×100 scale bar 100 μm, (G) NZ CD4-AP Foxp-3×100 scale bar 100 μm, (H) NZs CD4-AP Foxp-3×100 scale bar 100 μm, (I) NZm CD4-AP Foxp-3×100 scale bar 100 μm, (J) NZ MPO-AP×400 scale bar 100 μm, (K) NZs MPO-AP×400 scale bar 100 μm, (L) NZm MPO-AP×400 scale bar 100 μm.

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