Evaluation of co-therapy with melatonin and methylprednisolone in Experimental Autoimmune Encephalomyelitis (EAE) [Dataset]

Description

The dataset includes the raw data from the clinical score and flow cytometry analyzes carried out in the work titled " Melatonin synergistically potentiates the effect of methylprednisolone on reducing neuroinflammation in the experimental autoimmune encephalomyelitis mouse model of multiple sclerosis". This study shows the protective synergistic effect of co-treatment with melatonin and methylprednisolone on reducing the severity of EAE by decreasing CD4+ lymphocytes, B cells, macrophages and dendritic cells in the CNS, as well as modulating the population of infiltrated T and B cells toward regulatory phenotypes to the detriment of pro-inflammatory effector functions. In addition, treatment with melatonin from the clinical onset of EAE improves the natural course of the EAE and the response to a subsequent treatment with methylprednisolone in a later relapse of the disease. Eight-week-old female C57BL/6N mice were immunized immunization with 100 μg of MOG35–55 (Cambridge Research Biochemicals, Cleveland) emulsified in CFA (Sigma) containing 50 μg of heat-killed Mycobacterium tuberculosis (H37Ra, ATCC 25177) by subcutaneous injection in both hind legs and two doses of intraperitoneal pertussis toxin (200 ng/day) (List Labs, California) on days 0 and 2 post-induction. Animals were randomly divided to receive melatonin at a concentration of 80 mg/kg and/or methylprednisolone at a concentration of 40 or 160 mg/kg in different treatment regimens. Mice were sacrificed at the peak of the disease (day 15 after induction) and after perfusion, the CNS was collected, homogenized and enzymatically dissociated with 1.87 mg/ml of collagenase IV (Worthington) and 0.25 mg/ml of DNase I (AppliChem) for 35 min at 37°C to obtain a suspension of single cells. Subsequently, a 37%:70% discontinous percoll gradient was carried out to isolate CNS-infiltrating mononuclear cells. To assess the profile of infiltrated immune cells in the CNS, cells were stained for the following antibodies against surface markers: CD45, CD4, CD8α, CD19, CD11b, CD11c, CD44, CD62L, B220, CD138, PD-1 (CD279), CTLA-4 (CD152), FAS (CD95), and CD25. To identify Treg and analyze intracellular production of TNF, IFN-γ and IL-10, after surface staining, cells were fixed and permeabilized using the The dataset includes the raw data from the clinical score and flow cytometry analyzes carried out in the work titled " Melatonin synergistically potentiates the effect of methylprednisolone on reducing neuroinflammation in the experimental autoimmune encephalomyelitis mouse model of multiple sclerosis". This study shows the protective synergistic effect of co-treatment with melatonin and methylprednisolone on reducing the severity of EAE by decreasing CD4+ lymphocytes, B cells, macrophages and dendritic cells in the CNS, as well as modulating the population of infiltrated T and B cells toward regulatory phenotypes to the detriment of pro-inflammatory effector functions. In addition, treatment with melatonin from the clinical onset of EAE improves the natural course of the EAE and the response to a subsequent treatment with methylprednisolone in a later relapse of the disease. Eight-week-old female C57BL/6N mice were immunized immunization with 100 μg of MOG35–55 (Cambridge Research Biochemicals, Cleveland) emulsified in CFA (Sigma) containing 50 μg of heat-killed Mycobacterium tuberculosis (H37Ra, ATCC 25177) by subcutaneous injection in both hind legs and two doses of intraperitoneal pertussis toxin (200 ng/day) (List Labs, California) on days 0 and 2 post-induction. Animals were randomly divided to receive melatonin at a concentration of 80 mg/kg and/or methylprednisolone at a concentration of 40 or 160 mg/kg in different treatment regimens. Mice were sacrificed at the peak of the disease (day 15 after induction) and after perfusion, the CNS was collected, homogenized and enzymatically dissociated with 1.87 mg/ml of collagenase IV (Worthington) and 0.25 mg/ml of DNase I (AppliChem) for 35 min at 37°C to obtain a suspension of single cells. Subsequently, a 37%:70% discontinous percoll gradient was carried out to isolate CNS-infiltrating mononuclear cells. To assess the profile of infiltrated immune cells in the CNS, cells were stained for the following antibodies against surface markers: CD45, CD4, CD8α, CD19, CD11b, CD11c, CD44, CD62L, B220, CD138, PD-1 (CD279), CTLA-4 (CD152), FAS (CD95), and CD25. To identify Treg and analyze intracellular production of TNF, IFN-γ and IL-10, after surface staining, cells were fixed and permeabilized using the

Abstract

The data set titled "Mel+MPD co-therapy.xlsx" includes numerical data from flow cytometry analysis (first sheet) and clinical score analysis (second sheet). A legend is also incorporated into the third sheet detailing the different experimental groups of the study.

Abstract

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