Journal: Diabetes

Article Title: Loss of NADPH Oxidase–Derived Superoxide Skews Macrophage Phenotypes to Delay Type 1 Diabetes

doi: 10.2337/db14-0929

Figure Lengend Snippet: Pancreas-infiltrating macrophages from superoxide-deficient NOD.Rag mice display a decrease in TNF-α and IL-1β synthesis, and a concomitant increase in Arg-1 synthesis after diabetogenic CD4 T-cell transfer. Dot plots of TNF-α– (A), IL-1β–, and Arg-1– expressing (B) macrophages in the pancreata of NOD.Rag and NOD.Rag.Ncf1m1J recipient mice at 7 days post-transfer with NOD.BDC-2.5 CD4 T cells. Dot plots were generated by first gating on granulocytes and then gating on F4/80+ or CD11b+ cells via side scatter. For TNF-α and IL-1β, expression is shown by F4/80+, I-Ag7+ cells while Arg-1 expression by CD11b+, I-Ag7+ cells is displayed. Mean fluorescence intensity (MFI) values of TNF-α and IL-1β were quantified by F4/80+, I-Ag7+ cells, while those of Arg-1 were calculated from CD11b+, I-Ag7+ cells. Data are representative of four independent experiments.

Article Snippet: CD4 T cells were purified by negative selection according to the manufacturer’s instructions via the RoboSep CD4 T-cell isolation kit (STEMCELL Technologies) with >90% purity ( 17 ).

Techniques: Expressing, Generated, Fluorescence

Journal: Diabetes

Article Title: Loss of NADPH Oxidase–Derived Superoxide Skews Macrophage Phenotypes to Delay Type 1 Diabetes

doi: 10.2337/db14-0929

Figure Lengend Snippet: Pancreas-infiltrating macrophages from NOD.Rag mice treated with MnP display a decrease in TNF-α and a concomitant increase in Arg-1 synthesis after diabetogenic CD4 T-cell transfer. Dot plots of TNF-α– (A), IL-1β–, and Arg-1–expressing (B) macrophages in the pancreata of NOD.Rag recipient mice at 7 days post-transfer with NOD.BDC-2.5 CD4 T cells treated with HBSS or MnP. Dot plots were generated by first gating on granulocytes and then gating on F4/80+ or CD11b+ cells via side scatter. Mean fluorescence intensity (MFI) values of TNF-α and IL-1β were quantified by F4/80+, I-Ag7+ cells, while those of Arg-1 mean fluorescence intensity were calculated from CD11b+, I-Ag7+ cells. For TNF-α and IL-1β, expression is shown by F4/80+, I-Ag7+ cells, while Arg-1 expression by CD11b+, I-Ag7+ cells is displayed. Data are representative of four independent experiments.

Article Snippet: CD4 T cells were purified by negative selection according to the manufacturer’s instructions via the RoboSep CD4 T-cell isolation kit (STEMCELL Technologies) with >90% purity ( 17 ).

Techniques: Expressing, Generated, Fluorescence

Journal: medRxiv

Article Title: Elevation of cell-associated HIV-1 RNA transcripts in CSF CD4+ T cells, despite suppressive antiretroviral therapy, is linked to in vivo brain injury

doi: 10.1101/2021.12.22.21268288

Figure Lengend Snippet: ( a ) Range of recovered CSF cell counts for different CD45+ cells obtained in the pellets, including CD3+CD4+ and CD3+CD8+ T lymphocytes, CD14+ monocytes, CD19+ B cells and CD16+CD56+ NK cells; ( b ) Rate of detectability of cell free virus using single copy assay, in CSF and plasma, respectively and cell-associated (CA) HIV-1 RNA and DNA in CSF cells and PBMC, respectively; ( c ) comparison of HIV-1 CA-RNA copies, normalized per 10 6 CD4 T cells, for CSF cells and PBMC, respectively; ( d ) comparison of HIV-1 CA-DNA copies, normalized per 10 6 CD4 T cells, for CSF cells and PBMC, respectively; ( e ) numbers of CSF cells and PBMC used for extractions; ( f ) correlation of copy numbers of HIV-1 CA-RNA vs DNA in CSF cells; ( g ) correlation of copy numbers of HIV-1 CA-RNA vs DNA in CSF cells; and ( h ) correlation of copy numbers of HIV-1 CA-RNA in CSF cells vs HIV-1 CA-RNA in PBMC.

Article Snippet: CD14+ monocytes were isolated from cryopreserved PBMCs, which were thawed and resuspended in PBS with 2% FCS, using positive selection on the automated RoboSep platform (CD14+ cell positive separation kit, Cat No. 17858RF, StemCell Technology, Vancouver, Canada), according to the manufacturer’s directions.

Techniques:

Journal: medRxiv

Article Title: Elevation of cell-associated HIV-1 RNA transcripts in CSF CD4+ T cells, despite suppressive antiretroviral therapy, is linked to in vivo brain injury

doi: 10.1101/2021.12.22.21268288

Figure Lengend Snippet: ( a ) Rate of detectability of cell-associated (CA) HIV-1 RNA and DNA, respectively in highly purified CD14+ monocytes from PBMC; ( b ) number of highly purified CD14+ monocytes and number of CD4+ T cells from CD14-neg PBMC used for extractions for the Double R assay; ( c ) relative contribution of CD4 T cells and CD14+ monocytes to total number of CA HIV-1 RNA transcripts from PBMC; ( d ) comparison of CA HIV-1 RNA copy numbers in CD14+ monocytes from PBMC vs CD4 T cells in PBMC, in individual patient samples; ( e ) consistent results for CA HIV-1 RNA from CSF cells, PBMC CD4 T cells and PBMC CD14+ monocytes, respectively, for longitudinal follow up samples from two participants.

Article Snippet: CD14+ monocytes were isolated from cryopreserved PBMCs, which were thawed and resuspended in PBS with 2% FCS, using positive selection on the automated RoboSep platform (CD14+ cell positive separation kit, Cat No. 17858RF, StemCell Technology, Vancouver, Canada), according to the manufacturer’s directions.

Techniques: Purification