Journal: Animals : an Open Access Journal from MDPI

Article Title: The Odontocete Ear Canal-Associated Lymphoid Tissue (ECALT) and Lymph Nodes: Morphological and Pathological Description with Immuno-Phenotypic Characterisation

doi: 10.3390/ani12172235

Figure Lengend Snippet: The primary antibodies.

Article Snippet: The following primary antibodies were used: monoclonal mouse anti-human CD3 (Dako M7254), monoclonal rabbit anti-human CD20 (Thermo Scientific #RB-9013-P, Runcorn, UK), polyclonal rabbit anti-human Iba-1 (Wako #019-19741), monoclonal mouse anti-human HLA-Dr (Dako M0746, Glostrup, Denmark), monoclonal mouse anti-porcine Vimentin (clone V9, Dako M0725, Glostrup, Denmark), monoclonal mouse anti-human Cytokeratin (Dako M3515, Glostrup, Denmark), and monoclonal rabbit anti-human von Willebrand factor (factor VIII) (Dako A0082, Glostrup, Denmark) (for details, see ).

Techniques: Staining, Blocking Assay, Positive Control

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Odontocete Ear Canal-Associated Lymphoid Tissue (ECALT) and Lymph Nodes: Morphological and Pathological Description with Immuno-Phenotypic Characterisation

doi: 10.3390/ani12172235

Figure Lengend Snippet: The histological images: ( A ) HE staining; ( B ) CD3; ( C ) CD20; ( D ) HLA-DR; ( E ) Iba-1; ( F ) PanCK; ( G ) Vimentin of a mononuclear infiltrate (asterisks) between the left ear canal and cartilage in a bottlenose dolphin (444_L18). Scale bars = 50 µm.

Article Snippet: The following primary antibodies were used: monoclonal mouse anti-human CD3 (Dako M7254), monoclonal rabbit anti-human CD20 (Thermo Scientific #RB-9013-P, Runcorn, UK), polyclonal rabbit anti-human Iba-1 (Wako #019-19741), monoclonal mouse anti-human HLA-Dr (Dako M0746, Glostrup, Denmark), monoclonal mouse anti-porcine Vimentin (clone V9, Dako M0725, Glostrup, Denmark), monoclonal mouse anti-human Cytokeratin (Dako M3515, Glostrup, Denmark), and monoclonal rabbit anti-human von Willebrand factor (factor VIII) (Dako A0082, Glostrup, Denmark) (for details, see ).

Techniques: Staining

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Odontocete Ear Canal-Associated Lymphoid Tissue (ECALT) and Lymph Nodes: Morphological and Pathological Description with Immuno-Phenotypic Characterisation

doi: 10.3390/ani12172235

Figure Lengend Snippet: A histological image (274/18_8R) of the immunohistochemical analysis of a lymphoid follicle stained with ( A ) anti-CD3, ( B ) anti-CD20, ( C ) anti-HLA-DR, ( D ) anti-Iba1, ( E ) anti-vimentin ( E ). Scale = 100 µm.

Article Snippet: The following primary antibodies were used: monoclonal mouse anti-human CD3 (Dako M7254), monoclonal rabbit anti-human CD20 (Thermo Scientific #RB-9013-P, Runcorn, UK), polyclonal rabbit anti-human Iba-1 (Wako #019-19741), monoclonal mouse anti-human HLA-Dr (Dako M0746, Glostrup, Denmark), monoclonal mouse anti-porcine Vimentin (clone V9, Dako M0725, Glostrup, Denmark), monoclonal mouse anti-human Cytokeratin (Dako M3515, Glostrup, Denmark), and monoclonal rabbit anti-human von Willebrand factor (factor VIII) (Dako A0082, Glostrup, Denmark) (for details, see ).

Techniques: Immunohistochemical staining, Staining

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Odontocete Ear Canal-Associated Lymphoid Tissue (ECALT) and Lymph Nodes: Morphological and Pathological Description with Immuno-Phenotypic Characterisation

doi: 10.3390/ani12172235

Figure Lengend Snippet: The histological images of IHC in a striped dolphin nodular lymphoid tissue (ID362/18). ( A ) CD3; ( B ) CD20; ( C ) Iba-1; ( D ) vWf. Scale bar = 500 µm.

Article Snippet: The following primary antibodies were used: monoclonal mouse anti-human CD3 (Dako M7254), monoclonal rabbit anti-human CD20 (Thermo Scientific #RB-9013-P, Runcorn, UK), polyclonal rabbit anti-human Iba-1 (Wako #019-19741), monoclonal mouse anti-human HLA-Dr (Dako M0746, Glostrup, Denmark), monoclonal mouse anti-porcine Vimentin (clone V9, Dako M0725, Glostrup, Denmark), monoclonal mouse anti-human Cytokeratin (Dako M3515, Glostrup, Denmark), and monoclonal rabbit anti-human von Willebrand factor (factor VIII) (Dako A0082, Glostrup, Denmark) (for details, see ).

Techniques:

Journal: Cell Reports Medicine

Article Title: Heritable vaginal bacteria influence immune tolerance and relate to early-life markers of allergic sensitization in infancy

doi: 10.1016/j.xcrm.2022.100713

Figure Lengend Snippet:

Article Snippet: CD20 biotin Mouse anti-human , Thermo Fisher Scientific , Clone 2H7; Cat# 13-0209-82; RRID: AB_657690.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Blocking Assay, Staining, Lysis, Sequencing, Software, Amplification

Journal: Immunity

Article Title: Multimodal profiling of lung granulomas in macaques reveals cellular correlates of tuberculosis control

doi: 10.1016/j.immuni.2022.04.004

Figure Lengend Snippet:

Article Snippet: Rabbit anti-human CD20, polyclonal , ThermoFisher , Cat# RB-9013; RRID: AB_149767.

Techniques: Recombinant, Expressing, Software

Journal: Veterinary Sciences

Article Title: Molecular and Pathological Detection of Hepatitis E Virus in Roe Deer ( Capreolus capreolus ) and Fallow Deer ( Dama dama ) in Central Italy

doi: 10.3390/vetsci9030100

Figure Lengend Snippet: Antibodies, reagents used for antigenic unmasking, blockade of endogenous enzymes and non-specific protein bonds, dilution of the primary antibody, and type of secondary antibody used.

Article Snippet: Specific IHC was conducted with an anti-human CD3 rabbit polyclonal antibody (Dako, Glostrup, Denmark) diluted 1:200 directed against T-lymphocytes; with an anti-human CD20 rabbit polyclonal antibody diluted 1:100 (Thermo Fisher Scientific, Rockford, IL, USA) directed against the B-lymphocytes; with an anti-Iba-1 rabbit polyclonal antibody diluted 1:300 for the detection of macrophages.

Techniques: Blocking Assay

Journal: Veterinary Sciences

Article Title: Molecular and Pathological Detection of Hepatitis E Virus in Roe Deer ( Capreolus capreolus ) and Fallow Deer ( Dama dama ) in Central Italy

doi: 10.3390/vetsci9030100

Figure Lengend Snippet: Results of the immunohistochemical investigation conducted on the six liver samples that scored positive for RT-qPCR. 0 = absence; 1 = slight presence; 2 = moderate presence; 3 = strong presence.

Article Snippet: Specific IHC was conducted with an anti-human CD3 rabbit polyclonal antibody (Dako, Glostrup, Denmark) diluted 1:200 directed against T-lymphocytes; with an anti-human CD20 rabbit polyclonal antibody diluted 1:100 (Thermo Fisher Scientific, Rockford, IL, USA) directed against the B-lymphocytes; with an anti-Iba-1 rabbit polyclonal antibody diluted 1:300 for the detection of macrophages.

Techniques: Immunohistochemical staining

Journal: PLoS Pathogens

Article Title: CAR/CXCR5-T cell immunotherapy is safe and potentially efficacious in promoting sustained remission of SIV infection

doi: 10.1371/journal.ppat.1009831

Figure Lengend Snippet: Location of cell trace violet (CTV)-labeled infused T cells (pseudo-colored magenta) was determined in a chronically SIV-infected ART-naïve rhesus macaque, animal R14025 at two days post-treatment (DPT). CTV-labeled infused T cell product was detected in both the follicular and extrafollicular areas of lymphoid tissues. Representative images from spleen, lymph node (LN), bone marrow, ileum, lung, liver, brain, and rectum. Tissues were stained with anti-IgM or anti-CD20 (green) to label B cells and delineate B cell follicles (green). Arrowheads point to CTV-labeled cells. Scale bars = 100 μm.

Article Snippet: For immunofluorescence staining, sections were blocked with 4% normal goat serum (NGS) and incubated overnight with the following primary antibodies: mouse-anti-human CD20 (Clone L26, Biocare), mouse-anti-CD68 (KP1; Biocare), rabbit anti-CD20 (Polyclonal, Thermo Scientific), rabbit anti-CD3 (SP7; Labvision/Thermo Scientific), rabbit anti-CD4 (EPR6855; Abcam), or rabbit anti-Ki67 (Clone SP6, Invitrogen/Thermo Scientific).

Techniques: Labeling, Infection, Staining

Journal: PLoS Pathogens

Article Title: CAR/CXCR5-T cell immunotherapy is safe and potentially efficacious in promoting sustained remission of SIV infection

doi: 10.1371/journal.ppat.1009831

Figure Lengend Snippet: Location of CAR/CXCR5-T cells (red) within lymphoid tissues was determined in a chronically SIV-infected ART-naïve rhesus macaque, animal R14025 at 2 days post-treatment (DPT). Representative image of spleen tissue section showing duplex detection of CAR/CXCR5 construct (red) and SIV (pseudo-colored white) using RNAscope ISH combined with immunofluorescence using a custom-made probe for detection of CAR/CXCR5 construct and a probe specific for SIV. The white haze within the B cell follicle represents SIV virions trapped by the follicular dendritic cells (FDC) network. Scale bar = 100 μm. The right panels are enlargements showing an interaction between two CAR/CXCR5-transduced T cells and an SIV-infected cell. The tissue was stained with DAPI (blue), and anti-CD20 (green) to label B cells and delineate B cell follicles. Scale bar = 10 μm. Confocal images were collected using a 20× objective. The curves tool in Photoshop was used to increase the contrast of each image in a similar manner.

Article Snippet: For immunofluorescence staining, sections were blocked with 4% normal goat serum (NGS) and incubated overnight with the following primary antibodies: mouse-anti-human CD20 (Clone L26, Biocare), mouse-anti-CD68 (KP1; Biocare), rabbit anti-CD20 (Polyclonal, Thermo Scientific), rabbit anti-CD3 (SP7; Labvision/Thermo Scientific), rabbit anti-CD4 (EPR6855; Abcam), or rabbit anti-Ki67 (Clone SP6, Invitrogen/Thermo Scientific).

Techniques: Infection, Construct, Immunofluorescence, Staining

Journal: PLoS Pathogens

Article Title: CAR/CXCR5-T cell immunotherapy is safe and potentially efficacious in promoting sustained remission of SIV infection

doi: 10.1371/journal.ppat.1009831

Figure Lengend Snippet: CAR/CXCR5-T cells expanded at the edges of B cell follicles in vivo at 2 DPT and accumulated within B cell follicles at 6 DPT. (A) Representative image from LN tissue from Rh2850 stained 2 DPT showing CTV-labeled cell proliferation in the extrafollicular (EF) area near the follicular (F) zone. Tissues were stained with anti-CD3 (blue) to label T cells and anti-CD20 (green) to label B cells and delineate B cell follicles (F). The infused T cell product labeled with CTV is pseudo-colored magenta. Cells within F showed lower fluorescence intensity indicating division. Scale bar = 50 μm. (B) Representative image of LN tissue section, from Rh2858 visualized using RNAScope ISH combined with IF, showing expansion of CAR/CXCR5-T cells at the edge of follicles at 2 DPT. The right panel is an enlargement from the left panel showing a cluster of CAR/CXCR5-T cells (red) that appear to be expanding at the edge of the follicle. Tissues were stained with DAPI (blue) and anti-IgM (green) to label B cells and delineate B cell follicles (F), with the more brightly stained germinal center in the center of the F. (C) Representative image of LN tissue from Rh2850, showing CAR/CXCR5-T cell (red) proliferation in F and EF at 6 DPT detected by RNAScope ISH combined with IF. Tissues were stained with anti-IgM (Blue) and anti-Ki67 (green) to mark activation and proliferation. B cell follicles are delineated with white lines. Scale bars = 100 μm. Confocal images were collected using a 20× objective. (D) Percentage of Ki67 + CAR/CXCR5 T cells in the F (green) and EF (blue) areas for each of the T2 animals.

Article Snippet: For immunofluorescence staining, sections were blocked with 4% normal goat serum (NGS) and incubated overnight with the following primary antibodies: mouse-anti-human CD20 (Clone L26, Biocare), mouse-anti-CD68 (KP1; Biocare), rabbit anti-CD20 (Polyclonal, Thermo Scientific), rabbit anti-CD3 (SP7; Labvision/Thermo Scientific), rabbit anti-CD4 (EPR6855; Abcam), or rabbit anti-Ki67 (Clone SP6, Invitrogen/Thermo Scientific).

Techniques: In Vivo, Staining, Labeling, Fluorescence, Activation Assay

Journal: PLoS Pathogens

Article Title: CAR/CXCR5-T cell immunotherapy is safe and potentially efficacious in promoting sustained remission of SIV infection

doi: 10.1371/journal.ppat.1009831

Figure Lengend Snippet: CAR/CXCR5-T cells successfully homed to over 90% of the B cell follicles at 6 DPT and persisted for up to 28 DPT in SIV-infected ART-suppressed/released animals. Representative images of LN tissue section from Rh2858, showing CAR/CXCR5-T cells (red) detected using RNAScope ISH combined with IF using a custom-made probe for detection of the CAR/CXCR5 construct. (A) Confocal image showing a whole LN tissue section. (B) Enlargement of the delineated area in (6A) showing that CAR/CXCR5-transduced T cells (red) successfully homed to a B cell follicle (green). The tissue was stained with DAPI (blue), and anti-CD20 (green) to label B cells and delineate B cell follicles. The confocal image is collected with a 20× objective. Scale bar = 100 μm. (C) Levels of CAR/CXCR5-T cells over time after infusion in F (green) and EF areas (blue) of LN. The animal that lost control of the virus infection is marked with a red star. Samples not determined are marked ND. (D) Percentage of follicles that contained CAR/CXCR5-T cells over time post-infusion. (E) Frequency of CD4-MBL + cells in CD3 + PBMCs as determined by flow cytometry and (F) copies of CAR in the total cell population in PBMCs as determined by quantitative real-time PCR at the indicated time points post-infusion.

Article Snippet: For immunofluorescence staining, sections were blocked with 4% normal goat serum (NGS) and incubated overnight with the following primary antibodies: mouse-anti-human CD20 (Clone L26, Biocare), mouse-anti-CD68 (KP1; Biocare), rabbit anti-CD20 (Polyclonal, Thermo Scientific), rabbit anti-CD3 (SP7; Labvision/Thermo Scientific), rabbit anti-CD4 (EPR6855; Abcam), or rabbit anti-Ki67 (Clone SP6, Invitrogen/Thermo Scientific).

Techniques: Infection, Construct, Staining, Flow Cytometry, Real-time Polymerase Chain Reaction

Journal: PLoS Pathogens

Article Title: CAR/CXCR5-T cell immunotherapy is safe and potentially efficacious in promoting sustained remission of SIV infection

doi: 10.1371/journal.ppat.1009831

Figure Lengend Snippet: At 28 DPT, treated animals showed a reduction in viral (v)RNA compared to untreated control animals. (A) A representative image from a LN tissue section showing the abundance of SIV vRNA + cells and free virions trapped by follicular dendritic cells network (pseudo-colored white) detected using RNAscope ISH combined with IF in treated from Rh2850 (left panels) versus untreated control from R11002 (right panels). The tissue was stained with DAPI (blue), and anti-CD20 (green) to label B cells and delineate B cell follicles. The white haze within the B cell follicle represents SIV virions trapped by the follicular dendritic cells (FDC) network. Confocal images were collected with a 20× objective. Scale bar = 100 μm. (B) Levels of viral RNA in F and EF areas. The animal that lost viral control of the virus infection is marked with a red star. (C) The percentage of follicles with free virions bound by FDC.

Article Snippet: For immunofluorescence staining, sections were blocked with 4% normal goat serum (NGS) and incubated overnight with the following primary antibodies: mouse-anti-human CD20 (Clone L26, Biocare), mouse-anti-CD68 (KP1; Biocare), rabbit anti-CD20 (Polyclonal, Thermo Scientific), rabbit anti-CD3 (SP7; Labvision/Thermo Scientific), rabbit anti-CD4 (EPR6855; Abcam), or rabbit anti-Ki67 (Clone SP6, Invitrogen/Thermo Scientific).

Techniques: Staining, Infection

Journal: Cell Reports

Article Title: Cellular and humoral functional responses after BNT162b2 mRNA vaccination differ longitudinally between naive and subjects recovered from COVID-19

doi: 10.1016/j.celrep.2021.110235

Figure Lengend Snippet:

Article Snippet: Anti-human CD20 Pacific Orange (clone 2H7) , ThermoFisher Scientific , Cat# MHCD2030; RRID: AB_10375578.

Techniques: Recombinant, Staining, Enzyme-linked Immunosorbent Assay, Software

Journal: Frontiers in Immunology

Article Title: Epstein–Barr Virus+ B Cells in Breast Cancer Immune Response: A Case Report

doi: 10.3389/fimmu.2021.761798

Figure Lengend Snippet: CD3+ T cells (both CD4+ and CD8+) and CD20+ B cells were present and colocalized in the patient biopsy. Hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) from the triple-negative breast cancer (TNBC) core biopsy realized after diagnosis. (A) Histological section stain from breast carcinoma (HE ×200); (B) CD3 IHC stain in breast carcinoma highlighted numerous CD3+ T cells in the infiltrating front of neoplasia (×200); (C) CD20 IHC stain showed a similar distribution of CD20+ B cells compared with CD3+ T lymphocytes (×200); (D) CD4 IHC stain revealed that most lymphocytes are CD4+ T cells (×200); (E) CD8 IHC stain highlighted less CD8+ T cells but nearest to the tumor in the infiltrating front (×200).

Article Snippet: The primary antibodies used were rabbit polyclonal anti-human CD3 (Dako) and rabbit polyclonal anti-human CD20 (Thermo Fisher).

Techniques: Staining, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: Epstein–Barr Virus+ B Cells in Breast Cancer Immune Response: A Case Report

doi: 10.3389/fimmu.2021.761798

Figure Lengend Snippet: The tumor derived from a TNBC patient implanted in a PDX was a B-cell lymphocytic tumor (XABCL). CD20 immunohistochemical stain in the histological section revealed that most of the cells were CD20+ B cells. Cells with a bilobed nucleus, reminding of the Reed–Sternberg cells associated with Hodgkin’s lymphoma, were observed (pointed with a red arrow).

Article Snippet: The primary antibodies used were rabbit polyclonal anti-human CD3 (Dako) and rabbit polyclonal anti-human CD20 (Thermo Fisher).

Techniques: Derivative Assay, Immunohistochemical staining, Staining