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