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

Chiara Tani; Sabrina Vagnani; Linda Carli; Francesca Querci; Anja A Kühl; Simone Spieckermann; Constanze Pamela Cieluch; Simone Pacini; Rita Fazzi; Marta Mosca

doi:10.15283/ijsc17014

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

Int J Stem Cells. 2017 Nov 30;10(2):160-168. doi: 10.15283/ijsc17014.

Affiliations

¹ Rheumatology Unit, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
² Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, iPATH. Berlin, core unit of the Charité. Berlin, Germany.
³ Haematology Unit, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.

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 NZ_M 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.

Keywords: Animal model; Lupus nephritis; Mesenchymal stromal cells; Systemic lupus erythematosus.