Journal: Cell Reports Medicine

Article Title: Tissue-specific transcriptional profiles and heterogeneity of natural killer cells and group 1 innate lymphoid cells

doi: 10.1016/j.xcrm.2022.100812

Figure Lengend Snippet: Inter-tissue-specific signatures of NK and ILC1 subpopulations (A) Transcriptome-based clusters computed for each organ separately, projected onto the multi-organ integrated UMAP obtained with Harmony. (B) Heatmap representing overlap coefficient dissimilarity between transcriptome-based clusters calculated separately for each organ. (C) Dot plot of NK cell-specific versus ILC1-specific markers. Intermediate clusters were not taken into account. (D) Dot plot of specific markers for each cluster. (E) Flow cytometry profiles of CD49a, DNAM-1, PD-1H, and SDC4 expression, analyzed in Lin − NKp46 + NK1.1 + cells. Histograms represent the frequency of CD49a − CD49b + NK cells and CD49a + CD49b − ILC1s for each indicated marker across organs. Data are shown as mean ± SEM and are pooled from two independent experiments. Each point represents a pool of two mice.

Article Snippet: APC anti-mouse SDC4 (REA640) , Miltenyi Biotec , Cat# 130-109-831; RRID: AB_2653641.

Techniques: Flow Cytometry, Expressing, Marker

Journal: Cell Reports Medicine

Article Title: Tissue-specific transcriptional profiles and heterogeneity of natural killer cells and group 1 innate lymphoid cells

doi: 10.1016/j.xcrm.2022.100812

Figure Lengend Snippet:

Article Snippet: APC anti-mouse SDC4 (REA640) , Miltenyi Biotec , Cat# 130-109-831; RRID: AB_2653641.

Techniques: Recombinant, Saline, Staining, Cell Isolation, Sensitive Assay, Software

Journal: Brain Pathology

Article Title: Overexpression of the ubiquitin‐editing enzyme A20 in the brain lesions of Multiple Sclerosis patients: moving from systemic to central nervous system inflammation

doi: 10.1111/bpa.12906

Figure Lengend Snippet: List of antibodies used for neuropathological characterization of human post‐mortem snap‐frozen brain tissue by Immunohistochemistry (IHC) and immunofluorescence (IF). Abbreviations: MOG = myelin‐oligodendrocyte glycoprotein; MHC II = major histocompatibility complex class II molecules, GFAP = glial fibrillary acidic protein; NEFH = neurofilament heavy chain; PFA = paraformaldehyde.

Article Snippet: CD68 , Mouse , Monoclonal , 514H12 , IF 1:50 , 4% PFA , MCA1851T , AbD Serotec , macrophages.

Techniques: Immunohistochemistry, Immunofluorescence, Immunopeptidomics, Marker

Journal:

Article Title: Hepatitis C Virus-Like Particle Budding: Role of the Core Protein and Importance of Its Asp 111

doi: 10.1128/JVI.77.18.10131-10138.2003

Figure Lengend Snippet: Description of the SFV RNAs encoding HCV structural proteins and expression in BHK-21 cells. (A) RNAs transcribed from the four SFV DNA constructs. The SFV/Dj construct is our original Dj6.4 clone, encoding HCV core-E1-E2, and is described elsewhere (4). The horizontal line in each construct indicates the segment of the SFV genome. The position of the subgenomic SFV promoter is indicated (26S). Boxes show the inserted genes, and the shaded box in the SFV/Dj-C191 construct indicates the signal sequence targeting the E1 envelope glycoprotein to the ER. SPP, signal peptide peptidase processing between residues 173 and 174. The SFV/Dj-D111A construct was produced by introducing a D111-to-A substitution in the original SFV/Dj construct, i.e., introducing a viral L domain, PTAP. (B) Immunofluorescence staining, using a monoclonal anti-core antibody, of BHK-21 cells electroporated with SFV RNA β-Gal (a), Dj (b), Dj-C191 (c), Dj-C173 (d), and Dj-D111A (e). This assay was performed by following a standard procedure (35), using the mouse monoclonal anti-HCV core antibody MAb 1856 (Virostat, Portland, Maine). (C) Western blotting of these cells with monoclonal anti-core and anti-E2 antibodies. For this standard assay (4), the anti-core MAb 1856 gave poor results. We therefore carried out immunoblotting with the human monoclonal anti-core antibody B12.F8 (11) and the mouse monoclonal anti-E2 (10). Size markers (in kilodaltons) were obtained from New England Biolabs (Beverly, Mass.).

Article Snippet: This assay was performed by following a standard procedure ( 35 ), using the mouse monoclonal anti-HCV core antibody MAb 1856 (Virostat, Portland, Maine). (C) Western blotting of these cells with monoclonal anti-core and anti-E2 antibodies.

Techniques: Expressing, Construct, Sequencing, Produced, Immunofluorescence, Staining, Western Blot

Journal: Journal of Virology

Article Title: Novel Infectious cDNA Clones of Hepatitis C Virus Genotype 3a (Strain S52) and 4a (Strain ED43): Genetic Analyses and In Vivo Pathogenesis Studies ▿ †

doi: 10.1128/JVI.02667-09

Figure Lengend Snippet: Course of HCV infection and host immune responses in chimpanzee CH5276 following intrahepatic transfection with RNA transcripts of pS52 (genotype 3a). (A) Course of infection. Serum HCV RNA was monitored by in-house TaqMan assay (detection limit of 10 IU/ml) and/or by Roche Monitor test 2.0 (detection limit of 600 IU/ml). At the top of the panel, HCV RNA test results, anti-HCV antibody results, and liver histology scores are shown. Serum samples that were positive by TaqMan and/or by Monitor test 2.0 (solid black rectangles) and serum samples negative by TaqMan (white rectangle) are indicated in the top row. Anti-HCV antibodies were detected by 2nd generation ELISA: +, positive; −, negative. Necroinflammatory liver changes were scored as follows: 0, healthy or normal; 1, mild; 2, mild-moderate; 3, moderate-severe; 4, severe. HCV Monitor titers are indicated by the solid black circles in the graph; samples below the detection limit are shown as not detected (ND). The gray shaded area in the graph shows serum ALT levels (U/liter). The time points of direct sequence analysis of recovered viral genomes are indicated by the two white arrows. (B) Serum neutralizing antibodies. Percent neutralization of JFH1-based intergenotypic recombinants expressing S52 envelope proteins. The serum samples were diluted 1:20; >50% neutralization was considered significant. Values represent the means of three neutralizations; the standard error of the mean (SEM) range was 3 to 13%. Negative values are shown as 0%. For 1:80 serum dilutions, neutralization was <20% at all time points. (C and D) Peripheral and intrahepatic CD4+/CD8+ T-cell responses. The number of IFN-γ-secreting cells after stimulation with a panel of overlapping peptides specific for genotype 3a (strain K3a/650) and spanning the entire HCV polyprotein was determined in ELISPOT assays. PBMC were used directly. Intrahepatic CD4+ and CD8+ T cells were expanded from liver biopsy samples as described in Materials and Methods. The bars represent the total numbers of IFN-γ-secreting CD4+ and CD8+ T cells following stimulation with the different pools, after background subtraction. Cutoff points (dotted lines) were determined for individual experiments as described in Materials and Methods. Results below the cutoff are indicated by bars up to the dotted line. ND, not detectable.

Article Snippet: Huh7.5 cells were immunostained for HCV Core antigen using the primary mouse anti-HCV Core protein monoclonal antibody (B2) (Anogen, Yes Biotech Laboratories) at 1:200 in phosphate-buffered saline (PBS) with 5% bovine serum albumin and the secondary antibody Alexa Fluor 594-labeled goat anti-mouse IgG (Invitrogen) at 1:500 in PBS containing Tween; cell nuclei were counterstained with Hoechst 33342 (Invitrogen).

Techniques: Infection, Transfection, TaqMan Assay, Enzyme-linked Immunosorbent Assay, Sequencing, Neutralization, Expressing, Enzyme-linked Immunospot