Journal: Cells

Article Title: Current Paradigms of Tolerogenic Dendritic Cells and Clinical Implications for Systemic Lupus Erythematosus

doi: 10.3390/cells8101291

Figure Lengend Snippet: Clinical trials involving tolerogenic dendritic cells in SLE.

Article Snippet: Laquinimod (AhR agonist) , Teva Pharmaceutical industries/ NCT01085084 , July 2010–Dec 2012 , Phase II clinical trial in SLE with active lupus arthritis (N = 82) , No result posted yet , , .

Techniques: Expressing

Journal: Molecules

Article Title: Laquinimod Modulates Human Astrocyte Function and Dampens Astrocyte-Induced Neurotoxicity during Inflammation

doi: 10.3390/molecules25225403

Figure Lengend Snippet: Laq inhibits NFκB nuclear translocation and maintains glutamate transporters expression in astrocytes exposed to IL1β. ( A ) Chemical structures of laquinimod and de-laquinimod. ( B ) Representative immunofluorescence stainings for NFκB in human iAstrocytes exposed to drugs and/or IL1β. White arrows highlight positive nuclei. ( C ) The graph reports the percentage of NFκB positive nuclei under distinct conditions. ( D ) IL6 protein levels in supernatants from CTRL or IL1β-stimulated iAstrocytes after pre-exposure to drugs detected by the ELISA assay. ( E – H ) Representative immunofluorescence stainings for GLAST ( E ) and GLT1 ( G ) in human iAstrocytes and relative quantifications ( F , H ) under distinct conditions. For drug treatment, cells were exposed to 250 nM laquinimod or 100 nM de-laquinimod. DAPI was used for nuclear staining. Data are shown as mean ± SD of a representative experiment out of 2–3 independent experiments. Scale bars: 30 µm. # indicates statistical significance versus CTRL. * above the bar indicates statistical significance between specific experimental conditions. ** p < 0.01, *** p < 0.001, ## p < 0.01, ### p < 0.001.

Article Snippet: To detect NFkB activation and glutamate transporter expression, human iAstrocytes were eventually incubated with 10 µM CH223191 (Sigma, Milan, Italy) or vehicle (PBS or DMSO max 0.2% v/v) for 1 h, then exposed to 100 or 250 nM laquinimod (TEVA Pharmaceutical Industries, Rho, Milan, Italy), 100 nM de-laquinimod (TEVA Pharmaceutical Industries) or a vehicle for an additional 4 h, and finally stimulated with 10 ng/mL IL1β (Thermo Fisher Scientific) for 30 min for the NFκB assay or 24 h for the glutamate transporter assay.

Techniques: Translocation Assay, Expressing, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Staining

Journal: Molecules

Article Title: Laquinimod Modulates Human Astrocyte Function and Dampens Astrocyte-Induced Neurotoxicity during Inflammation

doi: 10.3390/molecules25225403

Figure Lengend Snippet: Laq activates nuclear AHR translocation but its effects on astrocyte functions are independent from AHR activity. ( A , C ) Representative immunofluorescence images for AHR in human iAstrocytes under distinct conditions and relative quantifications ( B , D ). ( E , F ) Immunofluorescence stainings for NFκB upon CH223191 treatment ( E ) and relative quantifications ( F ). ( G ) Images depicting GLAST stainings in human iAstrocytes and ( H , I ) frequency of GLAST ( H ) or GLT1 ( I ) highly expressing cells under the different experimental conditions. For drug treatment, cells were exposed to 250 nM laquinimod, 100 nM de-laquinimod and/or 10 µM CH223191. DAPI was used for nuclear staining. Data are shown as mean ± SD of a representative experiment out of 2–3 independent experiments. Scale bars: 30 µm. # indicates statistical significance versus CTRL. * above the bar indicates statistical significance between specific experimental conditions. * p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001.

Article Snippet: To detect NFkB activation and glutamate transporter expression, human iAstrocytes were eventually incubated with 10 µM CH223191 (Sigma, Milan, Italy) or vehicle (PBS or DMSO max 0.2% v/v) for 1 h, then exposed to 100 or 250 nM laquinimod (TEVA Pharmaceutical Industries, Rho, Milan, Italy), 100 nM de-laquinimod (TEVA Pharmaceutical Industries) or a vehicle for an additional 4 h, and finally stimulated with 10 ng/mL IL1β (Thermo Fisher Scientific) for 30 min for the NFκB assay or 24 h for the glutamate transporter assay.

Techniques: Translocation Assay, Activity Assay, Immunofluorescence, Expressing, Staining

Journal: Molecules

Article Title: Laquinimod Modulates Human Astrocyte Function and Dampens Astrocyte-Induced Neurotoxicity during Inflammation

doi: 10.3390/molecules25225403

Figure Lengend Snippet: Laq blocks neurodegeneration induced by astrocyte responses to IL1β. ( A ) Representative immunofluorescence stainings for DAPI (upper panels) and β-tubulin (lower panels) in neuronal cultures exposed to IL1β alone or pre-treated with Laq. ( B , C ) Quantification of cell number ( B ) and β-tubulin signal ( C ) expressed as the percentage with respect to control cultures. ( D ) Representative images showing DAPI (upper panels) and β-tubulin (lower panels) stainings in neuronal cultures exposure to distinct iAstrocyte-conditioned media and relative quantification ( E , F ). ( G – I ) Representative immunofluorescence images for neuronal cultures exposed to conditioned media from iAstrocytes stimulated in the presence of the AHR antagonist CH22319. For drug treatment, cells were exposed to 250 nM laquinimod, 100 nM de-laquinimod and/or 10 µM CH223191. Graphs show cumulative results from 2 independent experiments. Data are represented as mean ± SEM. Scale bars: 50 µm. # indicates statistical significance versus CTRL. * above the bar indicates statistical significance between specific experimental conditions. * p < 0.05, ** p < 0.01, *** p < 0.001, # p < 0.05, ## p < 0.01, ### p < 0.001.

Article Snippet: To detect NFkB activation and glutamate transporter expression, human iAstrocytes were eventually incubated with 10 µM CH223191 (Sigma, Milan, Italy) or vehicle (PBS or DMSO max 0.2% v/v) for 1 h, then exposed to 100 or 250 nM laquinimod (TEVA Pharmaceutical Industries, Rho, Milan, Italy), 100 nM de-laquinimod (TEVA Pharmaceutical Industries) or a vehicle for an additional 4 h, and finally stimulated with 10 ng/mL IL1β (Thermo Fisher Scientific) for 30 min for the NFκB assay or 24 h for the glutamate transporter assay.

Techniques: Immunofluorescence