Journal: Cell Reports Medicine

Article Title: Single-cell analysis shows that adipose tissue of persons with both HIV and diabetes is enriched for clonal, cytotoxic, and CMV-specific CD4+ T cells

doi: 10.1016/j.xcrm.2021.100205

Figure Lengend Snippet: Adipose tissue CD4 + T cell transcriptomes differ by HIV status, and CD4 + T cells are more commonly C-G-C + in HIV-positive versus CD69 + in HIV-negative persons (A) Adipose tissue CD4 + T cells from six HIV-positive and five HIV-negative diabetics were stained with the 12-antibody panel and analyzed by flow cytometry. The bar chart shows the distribution of adipose C-G-C + CD4 + T cells and CD69 + CD4 + T cells by HIV status. (B) We quantified (i) C-G-C + CD4 + T cells, (ii) CD69 + CD4 + T cells, and (iii) the combination of the two subsets. (C) scRNA-seq was performed on adipose tissue CD4 + memory T cells from HIV-positive (n = 2) versus HIV-negative diabetics (n = 5) on the same sequencing run, and differential gene expression was assessed using a panel of immune genes. (D) Gene enrichment pathway analysis was performed on differentially expressed genes higher in adipose tissue CD4 + T cells from HIV-positive diabetics with p < 0.05. Genes higher in HIV-negative diabetics did not enrich for Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways (data not shown). (E) Immunohistochemical stains of serial sections of perivascular adipose tissue show that CD4 + and CX3CR1 + cells are present in the adipose tissue of HIV-negative and HIV-positive diabetics. The p values were determined by Mann-Whitney U test and differential gene expression by Kruskal-Wallis test. Pathway enrichment analysis was performed using the web-based g:Profiler (version e100_eg47_p14_7733820, panel 1) and ShinyGO (v.0.61, panel 2). Analysis in g:Profiler was performed using the g:SCS method for multiple testing correction with an experiment-wide threshold of a = 0.05; ∗p < 0.05. See also and and .

Article Snippet: Perivascular Adipose Tissue (Formalin Fixed Paraffin Embedded) , CVPath Institute , https://www.cvpath.org/#OurResearchCapabilitiesandFacilities.

Techniques: Staining, Flow Cytometry, Sequencing, Gene Expression, Immunohistochemical staining, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Single-cell analysis shows that adipose tissue of persons with both HIV and diabetes is enriched for clonal, cytotoxic, and CMV-specific CD4+ T cells

doi: 10.1016/j.xcrm.2021.100205

Figure Lengend Snippet: Adipose tissue CD4 + T cells in HIV-positive diabetics have a cytotoxic RNA transcriptome signature (A) Bar plot and boxplot showing a higher combined proportion of CD69 + and C-G-C + CD4 + T cells in adipose tissue of HIV-positive diabetics (n = 6) versus non-diabetics (n = 5). (B) Differential gene expression of immune genes by adipose tissue CD4 + T cells from diabetics (n = 6, 354 cells) were compared with non-diabetics (n = 4, 273 cells). (C) Venn diagram showing overlap of genes with higher expression (p < 0.05) in matched adipose tissue and blood CD4 + T cells from diabetics versus non-diabetics. (D) GO processes enriched by overlapping genes differentially expressed by adipose and blood CD4 + T cells from diabetics. (E) Intracellular cytokine staining (ICS) showing the percentage of granzyme B + blood CD4 + T cells in HIV-positive non-diabetics (n = 15) and diabetics (n = 8) in unstimulated samples as well as co-expression of interferon γ (IFN-γ) and tumor necrosis factor alpha (TNF-α) with granzyme B after stimulation with staphylococcal enterotoxin B (SEB). (F and G) Percentage of IFN-γ and TNF-α co-expression by CD4 + T cells (F) and CD8 + T cells (G) after SEB and CMV-pp65 peptide pool stimulation. (H) Immunohistochemistry stains showing granzyme B + cells in perivascular adipose tissue of HIV-positive diabetics (n = 2) versus non-diabetics (n = 2). Differential gene expression was determined by Kruskal-Wallis test. Mann-Whitney U test was used to compare cytokine expression. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001. See also Figure S4 and .

Article Snippet: Perivascular Adipose Tissue (Formalin Fixed Paraffin Embedded) , CVPath Institute , https://www.cvpath.org/#OurResearchCapabilitiesandFacilities.

Techniques: Gene Expression, Expressing, Staining, Immunohistochemistry, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Single-cell analysis shows that adipose tissue of persons with both HIV and diabetes is enriched for clonal, cytotoxic, and CMV-specific CD4+ T cells

doi: 10.1016/j.xcrm.2021.100205

Figure Lengend Snippet:

Article Snippet: Perivascular Adipose Tissue (Formalin Fixed Paraffin Embedded) , CVPath Institute , https://www.cvpath.org/#OurResearchCapabilitiesandFacilities.

Techniques: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Recombinant, Library Quantification, Multiplex Assay, Sequencing, Labeling, Software

Journal: Acta Neuropathologica

Article Title: Familial globular glial tauopathy linked to MAPT mutations: molecular neuropathology and seeding capacity of a prototypical mixed neuronal and glial tauopathy

doi: 10.1007/s00401-019-02122-9

Figure Lengend Snippet: Characteristics of the antibodies used for immunohistochemistry and double-labelling immunofluorescence in human and mouse brain samples

Article Snippet: UCP5: mitochondrial uncoupling protein 5 , Rabbit polyclonal , 1:25 , Novus Biologicals/Bionova , Madrid, Spain.

Techniques: Immunohistochemistry, Immunofluorescence, Ubiquitin Proteomics

Journal: Acta Neuropathologica

Article Title: Familial globular glial tauopathy linked to MAPT mutations: molecular neuropathology and seeding capacity of a prototypical mixed neuronal and glial tauopathy

doi: 10.1007/s00401-019-02122-9

Figure Lengend Snippet: Gene symbols and TaqMan probes used in frontal cortex and subcortical white matter

Article Snippet: UCP5: mitochondrial uncoupling protein 5 , Rabbit polyclonal , 1:25 , Novus Biologicals/Bionova , Madrid, Spain.

Techniques:

Journal: Acta Neuropathologica

Article Title: Familial globular glial tauopathy linked to MAPT mutations: molecular neuropathology and seeding capacity of a prototypical mixed neuronal and glial tauopathy

doi: 10.1007/s00401-019-02122-9

Figure Lengend Snippet: Frontal cortex in GGT linked to MAPT P301T mutation (case 1). Double-labelling immunofluorescence and confocal microcopy of the frontal cortex (FC) in case 1 using antibodies anti-MPC1 ( a ), UCP4 ( b ), or UCP5 ( c ) (green), and anti-P-tau Thr181 or AT8 (red). MPC1 is expressed equally in astrocytes with tau deposits (thin arrows) and without tau deposits (thick arrow) ( a ). Similarly, UCP4 is found in astrocytes bearing phosphorylated tau (thin arrows) and in cells without tau deposits (thick arrows) ( b ). In contrast, UCP5 is absent in GAIs deposits (thin arrows) when compared with cells without tau deposits (thick arrows) ( c ). Nuclei are counterstained with DRAQ5™ (blue). Paraffin sections, a – c , bar = 30 μm. d – g Expression of histones in frontal cortex and white matter in GGT (case 1). H3K9me2 immunoreactivity is expressed equally in cells with (thin arrows) and without (thick arrows) phosphorylated tau deposits (P-tauThr181 antibody) ( d , e ). Similarly, H4K12ac immunoreactivity is expressed equally in oligodendrocytes with coiled bodies ( f ) and globular inclusions ( g ) (AT8 antibody) (thin arrows), and in oligodendroglia cells without tau inclusions (thick arrows). Nuclei are counterstained with DRAQ5™ (blue). Paraffin sections, d bar = 20 μm; e – g bar = 30 μm. h phosphorylated p38 Thr180-Tyr182 (p38P) co-localizes with AT8-immunoreactive deposits (asterisk) in frontal cortex in GGT. Paraffin section without nuclear counterstaining, bar = 10 μm

Article Snippet: UCP5: mitochondrial uncoupling protein 5 , Rabbit polyclonal , 1:25 , Novus Biologicals/Bionova , Madrid, Spain.

Techniques: Mutagenesis, Immunofluorescence, Expressing, Paraffin Section