Quality control and immunomodulatory potential for clinical-grade equine bone marrow-derived mesenchymal stromal cells and conditioned medium.

This study aimed to assess the safety and reproducibility of cell therapy for its use in clinical practice. We performed immunophenotypic characterization of equine bone marrow-derived mesenchymal stromal cells (BMMSCs) by flow cytometry using CD90, CD19, CD14, CD105, CD45, and HLA-DR markers (n = 4); GTG banding cytogenetic analysis (n = 3); and microbiological quality control (n = 4). The immunomodulatory potentials of BMMSCs (n = 4) and its conditioned medium (CM, n = 3) were investigated by in vitro lymphocyte inhibition assay using phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMCs). BMMSCs populations isolated from all animals showed high expression of CD90 and CD105, and low expression of CD19, CD4, CD45, and HLA-DR. Of the 60 metaphases analyzed, 5% presented aneuploidy on random chromosomes and no contamination was found based on microbiological analyses. Both treatments significantly inhibited lymphocyte proliferation (> 50%), compared with PHA-stimulated PBMCs (p < 0.0001). These promising results for BMMSCs and CM justify their potential as a therapeutic approach for inflammatory diseases. The techniques used in this study were effective in assessing the quality and determining the minimum criteria for the clinical use of BMMSCs in veterinary medicine.

Lung Tissue Damage Associated with Allergic Asthma in BALB/c Mice Could Be Controlled with a Single Injection of Mesenchymal Stem Cells from Human Bone Marrow up to 14 d After Transplantation.

Mesenchymal stem cell (MSC) research has demonstrated the potential of these cells to modulate lung inflammatory processes and tissue repair; however, the underlying mechanisms and treatment durability remain unknown. Here, we investigated the therapeutic potential of human bone marrow-derived MSCs in the inflammatory process and pulmonary remodeling of asthmatic BALB/c mice up to 14 d after transplantation. Our study used ovalbumin to induce allergic asthma in male BALB/c mice. MSCs were injected intratracheally in the asthma groups. Bronchoalveolar lavage fluid (BALF) was collected, and cytology was performed to measure the total protein, hydrogen peroxide (H2O2), and proinflammatory (IL-5, IL-13, and IL-17A) and anti-inflammatory (IL-10) interleukin (IL) levels. The lungs were removed for the histopathological evaluation. On day zero, the eosinophil and lymphochte percentages, total protein concentrations, and IL-13 and IL-17A levels in the BALF were significantly increased in the asthma group, proving the efficacy of the experimental model of allergic asthma. On day 7, the MSC-treated group exhibited significant reductions in the eosinophil, lymphocyte, total protein, H2O2, IL-5, IL-13, and IL-17A levels in the BALF, while the IL-10 levels were significantly increased. On day 14, the total cell numbers and lymphocyte, total protein, IL-13, and IL-17A levels in the BALF in the MSC-treated group were significantly decreased. A significant decrease in airway remodeling was observed on days 7 and 14 in almost all bronchioles, which showed reduced inflammatory infiltration, collagen deposition, muscle and epithelial thickening, and mucus production. These results demonstrate that treatment with a single injection of MSCs reduces the pathophysiological events occurring in an experimental model of allergic asthma by controlling the inflammatory process up to 14 d after transplantation.

Mineral supplement with pre- and probiotic improves respiratory immune status in yearling racing Thoroughbred horses: preliminary results / Suplemento mineral com pre-e probiótico melhora a condição imunológica respiratória em cavalos sobreano Puro Sangue Inglês de corrida: resultados preliminares

ABSTRACT: There are evidences of intestinal flora influence on the airway immune response. However, this information is scarce with regard to horses. The present preliminary study aimed to investigate the effects of a commercial mineral supplement containing probiotics and prebiotics on the respiratory immune response of horses supplemented with them during the first months of stabling and race training. Twelve TB yearlings were examined at the breeding farm at the beginning of the study (June 2016, henceforth M0) and then reexamined one (July 2016, henceforth M1) and five (November 2016, henceforth M2) months after being stabled at a racetrack in Curitiba, Brazil. Clinical and hematological examinations, airway endoscopy, and tracheal wash (TW) cytology were performed. After the initial examination, horses were randomly divided between the treatment group (TG), which received 10 grams of the supplement daily, and the control group (CG). There was no difference between the groups in terms of clinical parameters; in the hematological examination, eosinophil count was lower in the TG in M1 as compared with M0 (P = 0.045). Pharyngeal lymphoid hyperplasia (PLH) improved in the TG during M1 as compared with M0 (P = 0.007). However, on the CG, PLH remained unchanged. Relative number of eosinophils in the TW was 86.1% higher in M1 than M0 for the CG; in the TG; however, eosinophils were absent and remained within normal limits until M2. In conclusion, this study revealed a positive influence of probiotic and prebiotic supplementation on the respiratory immune response of young TB race horses. Further investigations are recommended.

RESUMO: As doenças respiratórias são uma grande preocupação em cavalos Puro Sangue Inglês (PSI) jovens durante o primeiro ano de treinamento para corrida. Existem evidências da influência da flora intestinal sobre a condição imunológica do organismo, incluindo a resposta de defesa das vias aéreas, porém essas informações são escassas nos cavalos. O presente estudo preliminar teve como objetivo investigar a influência da suplementação com núcleo mineral comercial com probiótico e prebiótico sobre a resposta imunológica respiratória de cavalos PSI durante os primeiros meses de estabulagem e treinamento. Doze cavalos PSI na fase de sobreano, machos e fêmeas, foram examinados ainda no haras (M0) e, um mês (M1) e cinco meses (M5) após estabulação no hipódromo em Curitiba, Brasil. Os cavalos foram submetidos a exames clínico e hematológico, endoscopia das vias aéreas e citologia do lavado traqueal (LT). Após o primeiro exame, os cavalos foram divididos aleatoriamente em grupo tratado (GT), que recebeu 10g por dia do suplemento, e grupo de controle (GC). Não houve diferença nos parâmetros clínicos, já no exame hematológico do GT, a contagem de eosinófilos foi menor em M1 quando comparado a M0 (P = 0,045). A hiperplasia folicular linfoide ficou dentro do limite normal e menores no GT em M1 quando comparado a M0 (P = 0.007), o que não ocorreu para o GC. O número relativo de eosinófilos no LT foi 86,1% maior em M1 versus M0 para o GC, enquanto no GT, os eosinófilos estavam ausentes e permaneceram dentro dos limites normais até M2. Concluindo, neste estudo preliminar, evidenciou-se uma influência positiva da suplementação probiótica e prebiótica na resposta imunológica respiratória de cavalos PSI jovens, justificando investigações adicionais.