Journal: Frontiers in Immunology

Article Title: Antagonizing FcαR1 (CD89) as treatment in IgA-mediated chronic inflammation and autoimmunity

doi: 10.3389/fimmu.2023.1118539

Figure Lengend Snippet: Anti-CD89 mAb 10E7 reduces neutrophil influx in a mouse model for linear IgA bullous disease. (A) Injection schedule of mice; Human IgA x human CD89 transgenic mice were injected 7 times with anti-mouse COLXVII human IgA in the right and PBS in the left ears. Treatment with anti-CD89 mAb clone 10E7 or the isotype mouse IgG1 control was given at day 7 and 11. (B) Neutrophil influx in PBS-injected ears (left panels) or human IgA anti-mouse COLXVII- injected ears (right panels) of human IgA x human CD89 transgenic mice that had been treated with anti-CD89 mAb 10E7 (upper panels) or isotype control (lower panels). Dapi (blue (in magnification) or grey (for overview ear)), GR-1 (green). (C) Quantification of neutrophil influx in ears. Mean ± SD are shown. Black circles and black squares represent number of neutrophils in PBS injected or anti-mouse collagen XVII human IgA injected ears of individual mice respectively. Mann-Whitney U tests,*; P<0.05, NS; not significant, mAb; monoclonal antibody, LABD; linear IgA bullous disease, COL; collagen, PBS; phosphate buffered saline, SD; standard deviation.

Article Snippet: Transgenic mice expressing human CD89 and knock-in for human IgA were subcutaneously (sc) injected with 10 μl (7mg/ml) human-anti-mouse Collagen XVII IgA (auto)antibodies (Amsterdam UMC and Polpharma Biologics Utrecht) in the right ear and 10 μl PBS in the left ear as control.

Techniques: Injection, Transgenic Assay, Control, MANN-WHITNEY, Saline, Standard Deviation

Journal: The Journal of Biological Chemistry

Article Title: Externalized Glycolytic Enzymes Are Novel, Conserved, and Early Biomarkers of Apoptosis *

doi: 10.1074/jbc.M111.314971

Figure Lengend Snippet: Underrepresented apoptotic membrane-associated proteins Proteins identified by iTRAQ analysis and found to be underrepresented among apoptotic membrane vesicles by at least 20% at a confidence level of >95% are listed.

Article Snippet: Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) ( A ) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) ( B ).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Externalized Glycolytic Enzymes Are Novel, Conserved, and Early Biomarkers of Apoptosis *

doi: 10.1074/jbc.M111.314971

Figure Lengend Snippet: Characterization of appearance of exposed glycolytic enzyme molecules. Murine splenocytes that were induced to undergo apoptosis with staurosporine were analyzed cytofluorometrically following staining with PE-conjugated annexin V, 7-AAD, and FITC-conjugated goat anti-EnoA peptide IgG polyclonal antibody (A), rabbit anti-GAPDH IgG polyclonal antibody and FITC-conjugated goat anti-rabbit IgG secondary antibody (B), and rabbit anti-TPI IgG polyclonal antibody and FITC-conjugated goat anti-rabbit IgG secondary antibody (C). Cells that met the criteria of staining positively with annexin V and negatively with 7-AAD (Annexin V+ 7-AAD−; the R1 region indicated in red in the upper dot plots) were gated electronically, and the fluorescein signal of those cells was analyzed (shown as solid green histograms in the lower panels). The fluorescein signal of annexin V+/7-AAD− cells stained with secondary antibody alone also is presented (gray dotted lines).

Article Snippet: Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) ( A ) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) ( B ).

Techniques: Staining

Journal: The Journal of Biological Chemistry

Article Title: Externalized Glycolytic Enzymes Are Novel, Conserved, and Early Biomarkers of Apoptosis *

doi: 10.1074/jbc.M111.314971

Figure Lengend Snippet: Cytofluorometric analysis of plasminogen binding to apoptotic cells. A, murine splenocytes that had undergone apoptosis spontaneously in culture (12 h) and freshly isolated, viable splenocytes were analyzed cytofluorometrically as described in the legend of Fig. 6 following staining with FITC-conjugated plasminogen. B, murine splenocytes that were induced to undergo apoptosis with staurosporine were analyzed cytofluorometrically following staining with PE-conjugated annexin V, 7-AAD, and FITC-conjugated plasminogen and analyzed as described in the legend of Fig. 8.

Article Snippet: Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) ( A ) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) ( B ).

Techniques: Binding Assay, Isolation, Staining

Journal: The Journal of Biological Chemistry

Article Title: Externalized Glycolytic Enzymes Are Novel, Conserved, and Early Biomarkers of Apoptosis *

doi: 10.1074/jbc.M111.314971

Figure Lengend Snippet: Cytofluorometric analysis of externalization of other molecules. Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) (A) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) (B). Apoptotic (solid green histograms) and viable (dashed lines) cells were identified by scatter properties and gated electronically.

Article Snippet: Human transformed (Jurkat) T lymphocytes that had been induced to undergo apoptosis by treatment with actinomycin D or that had been left untreated were analyzed cytofluorometrically following staining with FITC-conjugated mouse anti-human annexin II monoclonal antibody (BD Biosciences) ( A ) or mouse anti-calreticulin monoclonal and FITC-conjugated goat anti-rabbit IgG secondary antibodies (Enzo Life Sciences) ( B ).

Techniques: Transformation Assay, Staining