Este archivo ha sido creado el 20/07/2024 por Ana Isabel Álvarez-López GENERAL INFORMATION ------------------ 1. Dataset title: Evaluación de la co-terapia con melatonina y metilprednisolona en la Encefalomielitis Autoinmune Experimental (EAE) [Dataset] 2. Authorship: Name: Ana Isabel Álvarez-López Institution: Instituto de Biomedicina de Sevilla Email: aialvarez-ibis@us.es ORCID: 0000-0002-9408-0472 Name: Nuria Álvarez-Sánchez Institution: Instituto de Biomedicina de Sevilla Email: nalvarez-ibis@us.es ORCID: 0000-0002-1313-0440 Name: Ivan Cruz-Chamorro Institution: Universidad de Sevilla Email: icruz-ibis@us.es ORCID: 0000-0002-6547-8078 Name: Guillermo Santos-Sánchez Institution: Universidad de Sevilla Email: gsantos-ibis@us.es ORCID: 0000-0001-5790-2302 Name: Eduardo Ponce-España Institution: Universidad de Sevilla Email: eponce-ibis@us.es ORCID: 0000-0001-6197-0041 Name: Ignacio Bejarano Institution: Universidad de Sevilla Email: ibejarano@us.es ORCID: 0000-0003-3727-3873 Name: Patricia Judith Lardone Institution: Universidad de Sevilla Email: plardone@us.es ORCID: 0000-0003-1793-3985 Name: Antonio Carrillo-Vico Institution: Universidad de Sevilla Email: vico@us.es ORCID: 0000-0002-8516-0999 DESCRIPTION ---------- 1. Dataset language: Inglés 2. Abstract: 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 FoxP3/transcription factor staining buffer set (eBioscience) and stained with anti-FoxP3 or anti-TNF, -IFN-γ and -IL-10, respectively. The intracellular production of cytokines was carried out in cells cultured at 2.5 × 106 cells/ml with RPMI 1640 supplemented with 5% fetal bovine serum, 1% L-glutamine and 1% of penicillin/streptomycin incubated in the presence or absence of Phorbol-Myristate-Acetate and Ionomycin (PMAI, Sigma) with brefeldin A (eBioscience) for 5 h. Dead cells were excluded from analyses using the LIVE/DEAD® Fixable Dead Cell Stain Kit (Invitrogen). FACS analysis was performed using a Cytek Aurora spectral cytometer, and data were analyzed using FlowJo software (Treestar). 3. Keywords: Melatonin; Multiple Sclerosis; Methylprednisolone; EAE 4. Date of data collection (fecha única o rango de fechas): 03-05-2018 - 18-05-2022 5. Publication Date: 01-08-2024 6. Grant information: Grant Agency [Organismo financiador]: Fundación Progreso y Salud Grant Number [Código del proyecto]: PC-0019- 2017 Grant Agency [Organismo financiador]: Fundación Progreso y Salud Grant Number [Código del proyecto]: PI-0015-2018 Grant Agency [Organismo financiador]: Fundación Progreso y Salud Grant Number [Código del proyecto]: PI-0485-2014 Grant Agency [Organismo financiador]: Junta de Andalucía (Consejería de Economía y Conocimiento) Grant Number [Código del proyecto]: US-1263804 7. Geographical location/s of data collection: 37.36461597772077, -5.9786894560670305, Instituto de Biomedicina de Sevilla, Seville, Andalusia, Spain ACCESS INFORMATION ------------------------ 1. Creative Commons License of the dataset: CC BY-NC-ND 2. Dataset DOI: https://doi.org/10.12795/11441/161856 3. Related publication: Ana Isabel Álvarez-López, Nuria Álvarez-Sánchez, Ivan Cruz-Chamorro, Guillermo Santos-Sánchez, Eduardo Ponce-España, Ignacio Bejarano, Patricia Judith Lardone, Antonio Carrillo-Vico. Melatonin synergistically potentiates the effect of methylprednisolone on reducing neuroinflammation in the experimental autoimmune encephalomyelitis mouse model of multiple sclerosis. Journal of Autoimmunity. Volume 148, 2024, 103298, ISSN 0896-8411 4. Link to related datasets: Ana Isabel Álvarez-López, Nuria Álvarez-Sánchez, Cruz-Chamorro, I., Santos-Sánchez, G., Ponce-España, E., Bejarano, I., Patricia Judith Lardone, & Carrillo-Vico, A. (2024). Melatonin synergistically potentiates the effect of methylprednisolone on reducing neuroinflammation in the experimental autoimmune encephalomyelitis mouse model of multiple sclerosis. Journal of Autoimmunity, 148, 103298–103298. https://doi.org/10.1016/j.jaut.2024.103298 DOI/URL: https://doi.org/10.1016/j.jaut.2024.103298 VERSIONING AND PROVENANCE --------------- 1. Last modification date: 18-05-2022 2. Were data derived from another source?: No METHODOLOGICAL INFORMATION ----------------------- 1. Description of the methods used to collect and generate the data: To evaluate 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 FoxP3/transcription factor staining buffer set (eBioscience) and stained with anti-FoxP3 or anti-TNF, -IFN-γ and -IL-10, respectively. The intracellular production of cytokines was carried out in cells cultured at 2.5 × 106 cells/ml with RPMI 1640 supplemented with 5% fetal bovine serum, 1% L-glutamine and 1% of penicillin/streptomycin incubated in the presence or absence of Phorbol-Myristate-Acetate and Ionomycin (PMAI, Sigma) with brefeldin A (eBioscience) for 5 h. Dead cells were excluded from analyses using the LIVE/DEAD® Fixable Dead Cell Stain Kit (Invitrogen). FACS analysis was performed using a Cytek Aurora spectral cytometer. 2. Data processing methods: Data were analyzed using FlowJo software (Treestar). 3. Software or instruments needed to interpret the data: Graphics were designed using GraphPad Prism v8 software (GraphPad Software, Boston, MA, USA) 4. Information about instruments, calibration and standards: Citómetro Analizador Spectral Aurora Cytek (Beckman Coulter) Cuatro láseres. Arrays de detectores: Violeta: 405 nm 100mW. 16 APDs. Azul: 488nm 50mW. 14APDs. Verde: 561nm 50mW. 10 APDs. Rojo: 640 nm 80 mW. 8 APDs. FILE OVERVIEW ---------------------- 2. File list: File name: Mel+MPD_co-therapy.xlsx Description: 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. 4. File format: Excel (.xlsx) 5. If the dataset includes multiple files, specify the directory structure and relationships between the files: Not related files SPECIFIC INFORMATION FOR TABULAR DATA ------------------------------------------- 1. Name file: Mel+MPD_co-therapy.xlsx 2. Number of rows and columns: -1st sheet "Flow cytometry data": 235 rows and 35 columns -2nd sheet "Score data": 294 rows and 4 columns 3. Variables list: - 1st sheet "Flow cytometry data": Variable name: CD45high (% of total cells) Description: Total leukocytes infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CD4+ T lymphocytes (% of total cells) Description: CD4+ T lymphocytes infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CD8+ T lymphocytes (% of total cells) Description: CD8+ T lymphocytes infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CD19+ B lymphocytes (% of total cells) Description: B lymphocytes infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CD11b+ macrophages (% of total cells) Description: Macrophages infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CD11c+ dendritic cells (% of total cells) Description: Dendritic cells infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: TEM (% of CD4+ cells) Description: T Effector Memory cells in CD4+ infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: TNF+ (% of CD4+ cells) Description: CD4+ T lymphocytes producing TNF Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: IFN+ (% of CD4+ cells) Description: CD4+ T lymphocytes producing IFN Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: TNF+IFN+ (% of CD4+ cells) Description: CD4+ T lymphocytes producing TNF and IFN Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: TNF+ (% of TEM cells) Description: TEM cells producing TNF Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: IFN+ (% of TEM cells) Description: TEM cells producing IFN Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: TNF+IFN+ (% of TEM cells) Description: TEM cells producing TNF and IFN Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: PD-1+ (% of CD4+ cells) Description: CD4+ lymphocytes expressing PD-1 Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CTLA-4+ (% of CD4+ cells) Description: CD4+ lymphocytes expressing CTLA-4 Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: FAS+ (% of CD4+ cells) Description: CD4+ lymphocytes expressing FAS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CD25+FOXP3+ (% of CD4+ cells) Description: Treg infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: IL-10+ (% of CD4+ cells) Description: CD4+ T lymphocytes producing IL-10 Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: PD-1-IL-10- (% of CD8+ cells) Description: CD8 effector cells infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: TNF+ (% of CD8+PD-1-IL-10- cells) Description: CD8 effector cells producing TNF Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: IFN+ (% of CD8+PD-1-IL-10- cells) Description: CD8 effector cells producing IFN Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: PD-1+ (% of CD8+ cells) Description: CD8+ cells expressing PD-1 Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: IL-10+ (% of CD8+ cells) Description: CD8+ cells producing IL-10 Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: PD-1+IL-10+ (% of CD8+ cells) Description: CD8+ cells expressing PD-1 and producing IL-10 Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CD19+B220lowCD138+ plasmablasts (% of total cells) Description: Plasmablasts infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: CD19-B220lowCD138+ plasma cells (% of total cells) Description: Plasma cells infiltrating the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: TNF+ (% of CD19+ B cells) Description: B cells producing TNF Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: IL-10+ (% of CD19+ B cells) Description: B cells producing IL-10 Units of measure or value labels | Unidades de medida o etiquetas de valor: percentages Variable name: nº of total CD4+ cells Description: Total number of CD4+ cells infiltraing the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: Cell count Variable name: nº of total CD8+ cells Description: Total number of CD8+ cells infiltraing the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: Cell count Variable name: nº of total B cells Description: Total number of B cells infiltraing the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: Cell count Variable name: nº of total macrophages Description: Total number of macrophages infiltraing the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: Cell count Variable name: nº of total dendritic cells Description: Total number of dendritic cells infiltraing the CNS Units of measure or value labels | Unidades de medida o etiquetas de valor: Cell count - 2nd sheet "Score data": Variable name: Cumulative score Description: Cumulative score presented by the animals Units of measure or value labels | Unidades de medida o etiquetas de valor: Ordinal Scale Variable name: Maximum score Description: Maximum score presented by the animals Units of measure or value labels | Unidades de medida o etiquetas de valor: Ordinal Scale