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: Physiological Reports

Article Title: Depot and sex‐specific implications for adipose tissue expandability and functional traits in adulthood of late prenatal and early postnatal malnutrition in a precocial sheep model

doi: 10.14814/phy2.14600

Figure Lengend Snippet: Effects of prenatal and early postnatal nutrition on the adipocyte size distribution patterns in tissue slides of (a) subcutaneous (SUB), (b) epicardial (EPI), (c) perirenal (PER) and (d) mesenteric (MES) adipose tissue from male (♂) and female (♀) 2½ years old adult sheep. Values are expressed as the proportion (%) of adipocytes found in different cell size classes. Tissue slides were stained with Iron‐Hematoxylin except for EPI, which was stained with Hematoxylin and eosin for optimal staining of cell membranes. Cross‐sectional area of individual cells was determined in whole tissue scans using the Iron Haematoxylin Adipose Tissue software (APP ID 10,113; Visiopharm ® , Hoersholm, Denmark). NORM, HIGH, LOW, CONV and HCHF: See legends to Figure . Number of animals in the different groups: NORM ( N = 9, 3♂:6♀); HIGH ( N = 12, 5♂:7♀) and; LOW ( N = 15, 8♂:7♀). Number of animals in the early postnatal nutrition groups: CONV ( N = 20, 8♂:12♀) and HCHF ( N = 17, 8♂: 8♀). Ewed, Lambd, sex, Ewed*sex, Lambd*sex, Ewed*Lambd indicate significant effects found of the late gestation nutrition, early postnatal nutrition, sex of the sheep or their interactions, respectively

Article Snippet: The cell membrane, shape, and size of adipocytes were identified and measured using a specially designed morphometric application, Iron Hematoxylin Adipose Tissue (APP ID 10113; Visiopharm ® ; Figure ).

Techniques: Staining, Software