goat polyclonal anti cd4  (R&D Systems)

Journal: Biology

Article Title: Pilot Study of Anti-Th2 Immunotherapy for the Treatment of Breast Cancer-Related Upper Extremity Lymphedema

doi: 10.3390/biology10090934

Figure Lengend Snippet: Inclusion and exclusion criteria.

Article Snippet: Primary antibodies for immunohistochemical staining included rabbit polyclonal anti-human collagen I (ab34170; Abcam, Cambridge, UK), rabbit polyclonal anti-human Ki67 (ab15580; Abcam), goat polyclonal anti-human CD4 (AF379; R&D Systems, Minneapolis, MN, USA), goat polyclonal anti-human LYVE-1 (AF2089; R&D Systems), rabbit polyclonal anti-human periostin (ab14041; Abcam), rabbit polyclonal anti-human IL13 (ab9576; Abcam), mouse monoclonal anti-human mast cell tryptase (ab2378; Abcam), rabbit polyclonal anti-human thymic stromal lymphopoietin (TSLP; ab188766; Abcam), guinea pig polyclonal anti-human cytokeratin14 (ab192694; Abcam), goat polyclonal anti-human IL33 (AF3625; R&D System), rat monoclonal anti-IL25 (NBP2-11677; Novus Biologicals, Centennial, Colorado), rabbit polyclonal anti-human IL13 receptor (ab79277; Abcam), mouse monoclonal anti-human IL4 (MAB304; R&D Systems), and mouse monoclonal anti-human IL5 (MAB605; R&D Systems).

Techniques: Dissection, Bioprocessing

Journal: Biology

Article Title: Pilot Study of Anti-Th2 Immunotherapy for the Treatment of Breast Cancer-Related Upper Extremity Lymphedema

doi: 10.3390/biology10090934

Figure Lengend Snippet: Treatment with QBX258 decreases mast cell infiltration in lymphedematous skin. ( a ) Representative low-power (upper images) and high-power (lower images) immunofluorescent staining (left panels) and quantification (right panel) of DAPI (blue) and CD4 + cells (pink) in normal, LE, and LE + tx biopsy specimens. Area in dotted box is shown in high-power views. ( b ) Representative low-power (upper images) and high-power (lower images) immunofluorescent staining (left panels) and quantification (right panel) of DAPI (blue), LYVE-1 (red), and tryptase (green) in normal, LE, and LE + tx biopsy specimens. Area in dotted box is shown in high-power views. Scale bar (low-power images): 200 μm; scale bar (magnified images): 25 μm; * p < 0.05; ** p < 0.01.

Article Snippet: Primary antibodies for immunohistochemical staining included rabbit polyclonal anti-human collagen I (ab34170; Abcam, Cambridge, UK), rabbit polyclonal anti-human Ki67 (ab15580; Abcam), goat polyclonal anti-human CD4 (AF379; R&D Systems, Minneapolis, MN, USA), goat polyclonal anti-human LYVE-1 (AF2089; R&D Systems), rabbit polyclonal anti-human periostin (ab14041; Abcam), rabbit polyclonal anti-human IL13 (ab9576; Abcam), mouse monoclonal anti-human mast cell tryptase (ab2378; Abcam), rabbit polyclonal anti-human thymic stromal lymphopoietin (TSLP; ab188766; Abcam), guinea pig polyclonal anti-human cytokeratin14 (ab192694; Abcam), goat polyclonal anti-human IL33 (AF3625; R&D System), rat monoclonal anti-IL25 (NBP2-11677; Novus Biologicals, Centennial, Colorado), rabbit polyclonal anti-human IL13 receptor (ab79277; Abcam), mouse monoclonal anti-human IL4 (MAB304; R&D Systems), and mouse monoclonal anti-human IL5 (MAB605; R&D Systems).

Techniques: Staining

Journal: Science immunology

Article Title: Clonal expansion and activation of tissue-resident memory-like Th17 cells expressing GM-CSF in the lungs of severe COVID-19 patients.

doi: 10.1126/sciimmunol.abf6692

Figure Lengend Snippet: Fig. 2. T cell clonality in pulmonary inflammation. (A) Blood-lung activation map of T cells from blood and BALF of all patients: UMAP dimensionality reduction em- bedding of T cells (left); clone size proportion (clone count divided by number of cells per sample) of T cells (middle), and the cytokine secretion score of T cells (right) from COVID-19 and bacterial pneumonia as indicated. (B) UMAP presentation of T cells from BALF of all patients. Clusters were annotated according to gene expression and epitopes measurement of key markers. TCM, T central memory; TSCM, T stem cell–like memory; lncRNA, long noncoding RNA. (C) Ratio of clonal expansion of bacterial pneumonia versus COVID-19 for the major expanded BALF T cell clusters. (D) Subclustering analysis of clonally expanded CD4+ T cells of all patients. Clusters were anno- tated according to gene expression presented in the heatmap. (E) Volcano plot showing differential gene expression between TH17 clusters 1 and 2 of all patients. Genes were considered significant with adjust P < 0.05. Nonsignificant genes are shown in black. (F) Heatmap of selected pathogenic gene markers of TH17 cells of all patients in comparison with other T cell clusters. (G) Clone size proportion of T cells in peripheral blood and BALF of patients with COVID-19 and presentation of high abundant clones (clone size > 5) that are shared between BALF and blood and BAL-specific clones as indicated. (H) CD4 migration and tissue residency score of TH17 cluster1 (TRM17) and 2 (TEM17) from all patients. (I) Possible model of intraclonal diversification of CD4+ T cell subsets (left); distribution of two representative BALF clones from a patient with COVID-19 (patient S1 clone239 and clone218) on the UMAP (middle and right). (J) Bar plot of top expanded BALF clones containing TRM17 cells from patients with COVID-19. COVID-19: n = 8 for BALF and n = 7 for blood; bacterial pneumonia: n = 4 for BALF and n = 4 for blood.

Article Snippet: Immunofluorescence microscopy was performed in 1-m paraffin-embedded sections, after 15-min antigen retrieval with pH 9 antigen retrieval solution (Agilent, Santa Clara, CA, USA) and incubation with polyclonal primary goat anti-CD4 antibody (R&D Systems, Minneapolis, MN, USA, AF-379) and rabbit anti-CCR6 antibody (Abcam, Cambridge, UK, ab140768).

Techniques: Activation Assay, Gene Expression, Comparison, Clone Assay, Migration

Journal: Science immunology

Article Title: Clonal expansion and activation of tissue-resident memory-like Th17 cells expressing GM-CSF in the lungs of severe COVID-19 patients.

doi: 10.1126/sciimmunol.abf6692

Figure Lengend Snippet: Fig. 5. Cytokine secretion profile and cellular source of GM-CSF. (A) GM-CSF and IL-17A protein in serum of patients with COVD-19 (n = 8) and healthy controls (n = 7) from Hamburg and of patients with moderate (n = 8) or severe COVID-19 (n = 11) from Halle as indicated. Cell map of (B) CSF2 (GM-CSF) expressing and (C) IL17A express- ing cells (scale bars indicate normalized expression). Three different UMAPs with different cellular granularity showing the respective gene expression of in total cells of the BALF (left), total T cells of blood and BALF (middle), and total T cells in BALF (right) from all patients. (D) Immunofluorescence of CD4+ (green) CCR6+ (red) TRM17 cells in the lungs of a deceased patient with COVID-19 infection [nuclear staining 4′,6-diamidino-2-phenylindol (DAPI), blue] (two additional samples are presented in fig. S10B). (E) Combined immunofluorescence (CCR6) and FISH (IL17A) of lung samples from one patient with COVID-19. (F) Concentrations of the indicated cytokines in the BALF of patients with COVID-19 and bacterial pneumonia.

Article Snippet: Immunofluorescence microscopy was performed in 1-m paraffin-embedded sections, after 15-min antigen retrieval with pH 9 antigen retrieval solution (Agilent, Santa Clara, CA, USA) and incubation with polyclonal primary goat anti-CD4 antibody (R&D Systems, Minneapolis, MN, USA, AF-379) and rabbit anti-CCR6 antibody (Abcam, Cambridge, UK, ab140768).

Techniques: Expressing, Gene Expression, Immunofluorescence, Infection, Staining

Journal: Frontiers in Immunology

Article Title: The Reversal of Immune Exclusion Mediated by Tadalafil and an Anti-tumor Vaccine Also Induces PDL1 Upregulation in Recurrent Head and Neck Squamous Cell Carcinoma: Interim Analysis of a Phase I Clinical Trial

doi: 10.3389/fimmu.2019.01206

Figure Lengend Snippet: Tumors from patients treated with Tadalafil and MUC1/polyICLC vaccine show a lower infiltration of MDSCs and Treg and a higher infiltration of activated CD8 in the tumor bed. Computer based image cytometry was performed to enumerate the number of (A) MDSC, (B) IL4Rα − myeloid cells, (C) CD4 + T cell subsets, or (D) CD8 + T cells. (E) CD69 expression within the CD8 is reported normalized on the CD69 expression on all the cells evaluated. Depending on the region of interest evaluated, at least 10 5 -10 6 cells were analyzed. (F) The expression of CD69 in CD8 + T cells was plotted against MUC1 IHC score of the corresponding tumor. Two ways T -test and relevant pearson correlation parameters are reported.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-human FOXP3 antibody (clone 237/E7, dilution 1/25, Abcam) and the goat polyclonal anti-human CD4 antibody, (dilution 1/20, R&D System).

Techniques: Cytometry, Expressing

Journal: PLoS ONE

Article Title: FOXP3 Subcellular Localization Predicts Recurrence in Oral Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0071908

Figure Lengend Snippet: A) Immune fluorescence images 20X were taken on slides labeled with anti-CD4 (green) and anti-FOXP3 (red) antibodies and counterstained with DAPI for the nuclei. Images were segmented using Cellprofiler using the setting optimized to correctly segment lymphocytes as described in the material and methods. B) processedpicture showing the nuclei in a blue scale and the cytoplasm in red/yellow gradient. As shown in C), MFI values assume a “quasi” Gaussian distribution close to the 0. Empirically we found that a cell is considered positive by different investigators when its MFI is greater than a threshold (blue line) defined as: median (MFI)+1.7×(2XQ3-median). D) This method and this threshold were validated by counting CD4 + Foxp3 - (green circle), CD4 + nFOXP3 + cells (black triangle) or CD4 + cFOXP3 + (red triangle) cells in 20 pictures and by correlating those counts with the one obtained from the computer. The reported R 2 and equation correspond to the interpolated line when all the 3 cell types are included in the analysis. When CD4 + FOXP3 - , CD4 + cFOXP3 + or CD4 + nFOXP3 + are counted the R 2 values are 0.942, 0.956, and 0.890 respectively while the slope values are 0.983, 1.019, and 0.999 indicating a good correspondence between the “automatic” and the manual counting. E) CD4+FOXP3+ (regardless of the localization) were evaluated in recurrent or tumor free patients.

Article Snippet: Samples were incubated O/N at 4°C with the mouse monoclonal, anti-human FOXP3 antibody ab20034 (clone237/E7, dilution 1/25, Abcam) and the goat polyclonal anti-human CD4 antibodyAF-379-NK, (R&D bioscience, dilution 1/20) in PBS with 1% BSA.

Techniques: Fluorescence, Labeling

Journal: PLoS ONE

Article Title: FOXP3 Subcellular Localization Predicts Recurrence in Oral Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0071908

Figure Lengend Snippet: Tumor specimens werelabeled with anti-CD4 (green) and anti-FOXP3 (red) antibody and counterstained with DAPI (blue). Representative immune fluorescence microscopy photograph from ( A ) recurrent or ( C ) tumor free patients are shown. The corresponding images from the confocal microscope are shown in the inner panel. Different localization confirmed by confocal microscopy and examples of nuclear ( B ) or cytoplasmic ( D ) localization are shown. Images were analyzed by cell profiler and the percentage of CD4 + cells positive for FOXP3 only in the cytoplasm (cFOXP3), or only in the nucleus (nFOXP3), are reported respectively in E and F . Wilcoxon rank-sum test p value is reported.

Article Snippet: Samples were incubated O/N at 4°C with the mouse monoclonal, anti-human FOXP3 antibody ab20034 (clone237/E7, dilution 1/25, Abcam) and the goat polyclonal anti-human CD4 antibodyAF-379-NK, (R&D bioscience, dilution 1/20) in PBS with 1% BSA.

Techniques: Fluorescence, Microscopy, Confocal Microscopy

Journal: PLoS ONE

Article Title: FOXP3 Subcellular Localization Predicts Recurrence in Oral Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0071908

Figure Lengend Snippet: A) The percentage of CD4 + cFOXP3 + cells was plotted against the percentage of nFOXP3 + cells in tumor free (white circle) or recurrent (red circle) patients. Linear correlation between the two parameters was evaluated. B) Data from the percentage of FOXP3 positive cells in the nucleus or in the cytoplasm were transformed by adding 0.001 to 0 values and taking the ratioof nuclear to cytoplasmicFOXP3 + CD4 + tumor infiltrating T cells. Results are depicted after log 2 transformation with Wilcoxon rank sum test p -value. C) ROC plots. D) Univariate logistic regression and ROC analysis. OR : Odds ratio estimate for the increased risk of recurrence per twofold increase (log 2 ) or 50% decrease (−log 2 ) in marker shown in left hand column. CI : Confidence interval. AUC : area under the empirical receiver operating curve (ROC) shown in upper right panel.

Article Snippet: Samples were incubated O/N at 4°C with the mouse monoclonal, anti-human FOXP3 antibody ab20034 (clone237/E7, dilution 1/25, Abcam) and the goat polyclonal anti-human CD4 antibodyAF-379-NK, (R&D bioscience, dilution 1/20) in PBS with 1% BSA.

Techniques: Transformation Assay, Marker

Journal: PLoS ONE

Article Title: FOXP3 Subcellular Localization Predicts Recurrence in Oral Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0071908

Figure Lengend Snippet: Prognostic effect of the ratio of nuclear to cytoplasmic FOXP3 + CD4 + cells, adjusted for baseline characteristics.

Article Snippet: Samples were incubated O/N at 4°C with the mouse monoclonal, anti-human FOXP3 antibody ab20034 (clone237/E7, dilution 1/25, Abcam) and the goat polyclonal anti-human CD4 antibodyAF-379-NK, (R&D bioscience, dilution 1/20) in PBS with 1% BSA.

Techniques: