Adipose-derived mesenchymal stem cells ameliorate lung epithelial injury through mitigating of oxidative stress in mustard lung.

Aim: We investigated potential efficacy of autologous adipose-derived mesenchymal stem cell (MSC) on oxidative stress (OS) and airway remodeling in patients with chronic mustard lung. Patients & methods: Ten patients received 100 × 106 cells every 20 days for 4 injections over a 2-month period. Results: A gradual improvement was observed for 6 min walk test scores, pulmonary function tests and respiratory quality after MSCs therapy. A significant decrease was found for the mean levels of Mucin-1 protein (KL-6; p = 0.022) and Clara cell protein 16 (CC16; p = 0.005). Antioxidants had a tendency to be higher after each injection. Conclusion: Our findings revealed that MSCs therapy can be safely used for improvement of lung injury and regeneration in these patients without adverse effects. Trial registration number: NCT02749448 (ClinicalTrials.gov).

Safety and efficacy of stem cell therapy for treatment of neural damage in patients with multiple sclerosis.

Multiple sclerosis (MS) is a multifocal inflammatory disease that involves the central nervous system and associated with limbs paralysis and serious problems in sensation, limbs, visual and sphincter. This disease is a result of autoimmune mechanism in which autoantibodies target the self-myelin antigens and cause demyelination. Because of the myelin dysfunction, MS is clinically identified with neurological disabilities. Furthermore, it can be entered into the progressive phase because of irreversible neurodegeneration and axons damage. Unfortunately, there is no effective therapeutic method for this disease and current medications have been focused on amelioration of symptoms and chronic inflammation. Although current immunotherapies ameliorate the reactivity of autoimmune anti-myelin and MS relapse rate, there is no approved method for improvement of the disease progression and repairing of the damaged myelin. Therefore, finding an appropriate clinical treatment for improvement of neurological damages in MS patients is essential. Mesenchymal stem cells (MSCs) are multipotent cells with high proliferative and self-renewal capacities, as well as immunomodulatory and neuroregenerative effects. Bone marrow and adipose tissues derived MSCs have been considered for the treatment of different diseases because not only they can be easily isolated from these tissues, but also a patient can be served as a donor for himself without the risk of rejection. More importantly, autologous MSCs carry a safer pattern without the risk of malignant transformation. Here, we will discuss the effectiveness of MSCs therapy for MS patients by reviewing of clinical trials.

Stem cells therapy: a review on approaches that can be used for treatment of respiratory failures in sulfur mustard-injured patients.

Sulfur mustard (SM) is a toxic agent which causes severe abnormalities in an airway system such as necrosis and inflammation, oxidative stress, chronic bronchitis, shortness of breath, and chronic obstructive pulmonary disease. Although possible mechanisms of SM toxicity have been extensively considered, there is still need to find an appropriate clinical treatment to decrease chronic lung injuries caused by SM. Due to extensive progresses and achievement in tissue repairing through stem cells therapy, the importance of cell therapy for the treatment of lung injuries has been increased. However, several factors such as types of stem cells, necessary conditions for growth and proliferation of stem cells, and their homing into the target tissues are considered as the most important problems in this issue. Mesenchymal stem cells (MSCs) are a class of multipotent stem cells with proliferative and self-renewal capacity which are able to differentiate into different cell lines such as lung epithelial cells. They have a potential repairing and immune modulatory properties which make them as a good candidate for the regeneration of bronchioles tract in SM-exposed patients. Unlike chemical drugs, the differentiation and high-level safety properties of MSCs can be considered as a new strategy for the treatment of SM-injured patients with pulmonary complications. This review aims to consider the therapeutic effects of MSCs in the treatment of SM-induced pulmonary injuries in both animals and humans.

Adipose-Derived Mesenchymal Stem Cells for Treatment of Airway Injuries in A Patient after Long-Term Exposure to Sulfur Mustard.

OBJECTIVE: Sulfur mustard (SM) is a potent mutagenic agent that targets several organs, particularly lung tissue. Changes in morphological structure of the airway system are associated with chronic obstructive pulmonary deficiency following exposure to SM. Although numerous studies have demonstrated pathological effects of SM on respiratory organs, unfortunately there is no effective treatment to inhibit further respiratory injuries or induce repair in these patients. Due to the extensive progress and achievements in stem cell therapy, we have aimed to evaluate safety and potential efficacy of systemic mesenchymal stem cell (MSC) administration on a SM-exposed patient with chronic lung injuries. MATERIALS AND METHODS: In this clinical trial study, our patient received 100×106cells every 20 days for 4 injections over a 2-month period. After each injection we evaluated the safety, pulmonary function tests (PFT), chronic obstructive pulmonary disease (COPD) Assessment Test (CAT), St. George's Respiratory Questionnaire (SGRQ), Borg Scale Dyspnea Assessment (BSDA), and 6 Minute Walk Test (6MWT). One-way ANOVA test was used in this study which was not significant (P>0.05). RESULTS: There were no infusion toxicities or serious adverse events caused by MSC administration. Although there was no significant difference in PFTs, we found a significant improvement for 6MWT, as well as BSDA, SGRQ, and CAT scores after each injection. CONCLUSION: Systemic MSC administration appears to be safe in SM-exposed patients with moderate to severe injuries and provides a basis for subsequent cell therapy investigations in other patients with this disorder (Registration Number: IRCT2015110524890N1).

Immunomodulatory Properties of Mesenchymal Stem Cells Can Mitigate Oxidative Stress and Inflammation Process in Human Mustard Lung.

Oxidative stress and inflammation are one of the main pathological consequences of sulfur mustard on human lungs. Unfortunately, there is no effective treatment to mitigate pathological effects of sulfur mustard in mustard lungs. Here, we aimed to evaluate potential efficacy of systemic mesenchymal stem cells administration on expression of oxidative stress- and inflammation-related genes in sulfur mustard-exposed patients. Our patient received 100 million cells per injection, which was continued for four injections within 2 months. Sputum samples were provided after each injection. Oxidative stress was evaluated by determining sputum levels of malondialdehyde and glutathione. Furthermore, changes in expression of several oxidative stress- (metallothionein 3, glutathione reductase, oxidative stress responsive 1, glutathione peroxidase 2, lacto peroxidase, forkhead box M1) and inflammation-related genes (matrix metallopeptidase 2, matrix metallopeptidase 9, transforming growth factor-ß1, vascular endothelial growth factor, metallopeptidase inhibitor 1, metallopeptidase inhibitor 2) were also evaluated using real-time PCR after treatments. Two-lung epithelial-specific proteins including Clara cell protein 16 and Mucin-1 protein levels were measured using enzyme immunoassay method. No significant differences were found between serum levels of Clara cell protein 16 and serum Mucin-1 protein in patient before and after cell therapy. Most of the oxidative stress responsive genes, particularly oxidative stress responsive 1, were overexpressed after treatments. Expressions of antioxidants genes such as metallothionein 3, glutathione reductase and glutathione peroxidase 2 were increased after cell therapy. Upon comparison of inflammation-related genes, we observed upregulation of vascular endothelial growth factor and matrix metallopeptidase 9 after mesenchymal stem cells therapy. Additionally, a trend for increased value of glutathione and decreased levels of malondialdehyde was observed from baseline to final evaluation times. Mesenchymal stem cells administration mitigates oxidative stress and inflammation in sulfur mustard-exposed patients.

Therapeutic Potential of Mesenchymal Stem Cells for the Treatment of Airway Remodeling in Pulmonary Diseases.

According to significant improvements in the tissue engineering field over the past several years, lung tissue cells have recently attracted more attention due to the high prevalence and diversity in related diseases. However, selection of an appropriate cell type, screening of suitable conditions for growth and proliferation, as well as subsequent implantation into the body to repair and regenerate damaged tissues are considered as important issues in this context. It should also be noted that most studies have been described in animal models, but not in humans. Because of the high regenerative capacity, predominant immunomodulatory feature, and inhibition of T-lymphocyte proliferation, mesenchymal stem cells (MSCs) may play an important role in the reconstruction of damaged tissues including bronchioles in pulmonary diseases. Interestingly, clinical trial studies demonstrated that MSCs have the significant potential to treat a wide variety of diseases including acute myocardial infarction (AMI), liver cirrhosis, crohn's disease, and graft-versus-host disease (GVHD).

Monoclonal antibodies against 24 kDa surface antigen of hepatitis B viruses.

Hepatitis B virus (HBV) infection is one of the major public health problems worldwide. Effective control of HBV transmission in areas of high and intermediate endemicity would not be possible without vaccination of the vulnerable group. The diagnosis of acute and chronic hepatitis B infection is based on the detection of hepatitis B surface antigen (HBsAg). In this study, we prepared nine hybridomas that produced monoclonal antibodies specific for HBsAg. BALB/c mice were immunized with 24 kDa HBsAg and the immune spleen cells were fused with SP2/0 myeloma cell line. We obtained seven IgG1, one IgM, and one IgG2b positive clones. The stable hybrids were sub-cloned and ascitic fluid was prepared in the BALB/c mice. After antibody purification by protein G, the affinity column was prepared and the pure 24 kDa HBsAg from cell extract was eluted from the column. Western blot analysis showed that all monoclonal antibodies are specific for 24 kDa antigen. Since 24 kDa HBsAg is used for vaccination against hepatitis, these monoclonal antibodies are the best candidate for the isolation and purification of recombinant HBsAg from yeast expression vector by affinity column to be used for vaccination.

Epitope mapping of a common 57 kDa antigen of Leishmania species by monoclonal antibodies.

BALB/c mice were immunized with freeze-thawed promastigote of Leishmania infantum. Five monoclonal antibodies (mAb) were selected, four IgM (designated as P1A9, P2G8, P5E3 and P6B3) and one IgG1 (P3D2). ELISA and Western blot analysis suggested that all monoclonal antibodies are specific to a band of 57 kDa antigen of L. infantum as well as other three Leishmania species (L. tropica, L. major and L. donovani). ELISA additivity tests revealed four epitopes on 57 kDa antigen as defined by four IgM monoclonal antibodies. Three distinct epitopes were recognized by P1A9, P2G8, and P6B3 antibodies and one epitope recognized by P5E3 antibody that shared with P2G8, and P6B3 epitopes. The 57 kDa protein was purified with affinity column and was shown to possess proteolytic activity. It seems that 57 kDa protein is the major surface Leishmania antigen (gp63) that has been used as subunit vaccine with appropriate adjuvant and induced protection against L. major infection in BALB/c mice.

Production and characterization of monoclonal antibodies recognizing a common 57-kDa antigen of Leishmania species.

BACKGROUND: The therapy of leishmania infection is difficult and each year 1.5 million new cases of cutaneous leishmaniasis and 500,000 new cases of visceral leishmaniasis are estimated, therefore, there is a need for an effective vaccine. Monoclonal antibody (mAb) is one of the suitable methods for isolation and purification of leishmania antigens. In this report, we produced several mAb against leishmania infantum antigens for antigen purification to be used as candidate vaccine. METHODS: BALB/c mice were injected with freeze-thawed promastigote twice together with Freund adjuvant. Three days before fusion, antigen in saline was injected into the tail vain and then mice were killed and the spleen lymphocytes were fused with myeloma SP2/0. RESULTS: Five mAb against promastigote form of Leishmania infantum parasite were obtained. Western-blot analysis showed that these mAb recognize a band of 57- kDa protein either in parasite lysate or on whole L. infantum, L. tropica, L. major and L. donovani. It seems that the 57 kDa-protein is the major surface leishmania antigen (gp63) that is neither stage-specific nor differentially regulated. These mAb do not recognize the recombinant gp63 antigen and seems recognizing only the native form of a gp63 isoform. The IgG1 mAb was purified by affinity column and was used to purify 57 kDa antigens from Leishmania lysate. CONCLUSION: Since these antibodies recognizing one specific protein band in 4 different strains of leishmania, they could be used for leishmania diagnostic kits and also for purification of antigen to be tested for its protective effect against leishmania infection.