Journal: Journal of neuroimmunology

Article Title: Corticotropin-releasing hormone receptor-1 and 2 activity produces divergent resistance against stress-induced pulmonary Streptococcus pneumoniae infection

doi: 10.1016/j.jneuroim.2011.06.016

Figure Lengend Snippet: Administration of antagonist resulted in changes of BALF IL-17A cytokine production. IL-17A cytokine production was determined from BALF of mice (n=5) per group by ELISA. Group differences were analyzed by one-way ANOVA. Asterisk (*) indicates significant (p≤0.05) difference between all experimental groups.

Article Snippet: ELISA antibody set and recombinant cytokine for standard were purchased from R&D Systems (R&D Systems Inc. Minneapolis, MN) for recombinant IL-17A.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cardiovascular Medicine

Article Title: Inhibition of IL17A Using an Affibody Molecule Attenuates Inflammation in ApoE-Deficient Mice

doi: 10.3389/fcvm.2022.831039

Figure Lengend Snippet: In vitro efficacy of Affibody molecules in blocking IL17A induced responses in human aortic smooth muscle cells (SMCs) and mouse NIH/3T3 fibroblast cells. ELISA data showing the release of CXCL1 and IL6 into the medium (24 h) induced by increasing concentration of recombinant IL17A in human SMCs ( n = 4) (A,B) and CXCL1 in mouse 3T3 fibroblast cells ( n = 4) (E) . Human SMCs were stimulated with 5 ng/ml recombinant IL17A and without or with increasing concentrations of human Affibody molecules against IL17A and release of CXCL1 and IL6 (24 h) was assessed by ELISA ( n = 3) (C,D) . Mouse NIH/3T3 fibroblast cells were stimulated with 25 ng/ml recombinant IL17A and without or with increasing concentration of Affibody molecule against mouse IL17A and release of CXCL1 (24 h) was assessed by ELISA ( n = 5) (F) . Data is presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control or as mentioned in graph (vs. IL17A).

Article Snippet: The next day, the media was replaced with fresh antibiotics free medium, and cells were treated with different concentrations of human or murine recombinant IL17A (R&D systems, USA) and cultured in the presence or absence of human or murine Affibody molecule at different concentrations to block IL17A (Affibody AB, Sweden), for 24 h. Unstimulated cells served as reference.

Techniques: In Vitro, Blocking Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Recombinant, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Inhibition of IL17A Using an Affibody Molecule Attenuates Inflammation in ApoE-Deficient Mice

doi: 10.3389/fcvm.2022.831039

Figure Lengend Snippet: Effect of Affibody molecule targeting IL17A in atherosclerotic lesion development in ApoE −/− mice. Quantification of lesion size (%) in the aortic arch (A) and brachiocephalic artery (B) of control mice and αIL17A Affibody molecule treated mice. Quatification of lesion areas (μm 2 ) in the aortic root (C) . Representative Oil Red O immunostainings from the aortic root of control (D) and mice treated with αIL17A Affibody molecule (E) . Data are presented as Median with IQR.

Article Snippet: The next day, the media was replaced with fresh antibiotics free medium, and cells were treated with different concentrations of human or murine recombinant IL17A (R&D systems, USA) and cultured in the presence or absence of human or murine Affibody molecule at different concentrations to block IL17A (Affibody AB, Sweden), for 24 h. Unstimulated cells served as reference.

Techniques: Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Inhibition of IL17A Using an Affibody Molecule Attenuates Inflammation in ApoE-Deficient Mice

doi: 10.3389/fcvm.2022.831039

Figure Lengend Snippet: Blood and morphometric parameters ^a and T cell subsets in spleen ^b .

Article Snippet: The next day, the media was replaced with fresh antibiotics free medium, and cells were treated with different concentrations of human or murine recombinant IL17A (R&D systems, USA) and cultured in the presence or absence of human or murine Affibody molecule at different concentrations to block IL17A (Affibody AB, Sweden), for 24 h. Unstimulated cells served as reference.

Techniques: Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Inhibition of IL17A Using an Affibody Molecule Attenuates Inflammation in ApoE-Deficient Mice

doi: 10.3389/fcvm.2022.831039

Figure Lengend Snippet: Affibody molecule against IL17A reduces plasma levels of inflammatory and/or atherosclerosis-associated proteins in ApoE −/− mice. The Volcano plot shows fold changes (FC) and p -values of differentially altered plasma proteins analyzed by olink proteomics, in mice treated with αIL17A Affibody molecule ( n = 11) compared to sham ( n = 12) (A) . The p -values are presented on log10 scale while the FCs were calculated from linearized normalized protein expression (NPX) values and presented on log2 scale. Positive log2 fold change values correspond to higher protein levels and negative values correspond to reduced protein levels in mice treated with αIL17A Affibody molecule. The horizontal dotted line reflects the cut-off for statistical significance ( p < 0.05). Blue circles highlight proteins with a significant decrease ( p < 0.05) and red circles highlight proteins with a significant increase ( p < 0.05). Gray circles represent proteins with no statistical significance. Correlation between the plasma concentration of CXCL1 protein and the area of aortic root lesion in atherosclerotic prone ApoE −/− mice ( n = 8) (B) .

Article Snippet: The next day, the media was replaced with fresh antibiotics free medium, and cells were treated with different concentrations of human or murine recombinant IL17A (R&D systems, USA) and cultured in the presence or absence of human or murine Affibody molecule at different concentrations to block IL17A (Affibody AB, Sweden), for 24 h. Unstimulated cells served as reference.

Techniques: Expressing, Concentration Assay

Journal: Frontiers in Cardiovascular Medicine

Article Title: Inhibition of IL17A Using an Affibody Molecule Attenuates Inflammation in ApoE-Deficient Mice

doi: 10.3389/fcvm.2022.831039

Figure Lengend Snippet: Effects of Affibody molecule against IL17A on gene expression in splenocytes and thoracic aorta from ApoE −/− mice. mRNA levels were assessed by quantitative real-time reverse transcription polymerase chain reaction (qPCR) analysis. Analysis of Cxcl1, Il6, Ccl20 , and Vcam1 genes in splenocytes from sham ( n = 6) and αIL17A Affibody molecule treated mice ( n = 5) (A-D) . Analysis of Casp3, Cd3e Cxcl1 , and Il6 transcripts in the thoracic aorta from sham ( n = 12) and αIL17A Affibody molecules treated mice ( n = 10-11) (E-H) . Data are presented as Median with IQR. NS, Not significant.

Article Snippet: The next day, the media was replaced with fresh antibiotics free medium, and cells were treated with different concentrations of human or murine recombinant IL17A (R&D systems, USA) and cultured in the presence or absence of human or murine Affibody molecule at different concentrations to block IL17A (Affibody AB, Sweden), for 24 h. Unstimulated cells served as reference.

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction