anti itga7 alexa fluor 700  (R&D Systems)

Journal: Nature Medicine

Article Title: Innate antiviral and immune functions associated with the HIV reservoir decay after anti-PD-1 therapy

doi: 10.1038/s41591-025-04139-y

Figure Lengend Snippet: a , Ridge regression model weights of features associated with anti-PD-1 therapy at baseline (C01D01) and 24 hours after therapy (C01D02) that correlated with frequencies of HIV DNA-containing cells at EOT. b , Correlation between predicted and actual HIV DNA levels per million CD4 + T cells based on the ridge model ( ρ = 0.59, P = 0.035). c , Model illustrating cell-subset-specific ISG expression. The left heatmap shows ISG expression across monocyte, CD4 + and CD8 + T cell subclusters (red, high; gray, low). The middle bar plots display the relative ( z -scaled) fold change in gene expression at C01D02 versus C01D01 (values >0 indicate upregulation). The right heatmap shows expression of monocyte-specific and T-cell-specific ISGs at EOT in group 1 versus group 2 participants, normalized per gene (row z -score; red, increased expression; blue, decreased expression). Bar plots above indicate total HIV DNA levels per participant at EOT. d , In vitro stimulation assay. Memory CD4 + T cells and CD8-depleted PBMCs from five donors ( n = 5) were cultured with or without innate immune stimuli (IFNα, IFNγ, poly I:C or R848). Mean fluorescence intensity (MFI) of the antiviral proteins APOBEC3G and IFIT1 in memory CD4 + T cells was quantified by flow cytometry. Each point represents a biological replicate (donor); bars denote the mean ± s.e.m. Comparisons between memory CD4 + -only cultures and CD8-depleted PBMC co-cultures for each condition were performed using paired t -tests. e , f , In vitro HIV challenge assay. Memory CD4 + T cells and CD8-depleted PBMCs from five donors ( n = 5) were infected with HIV in the presence or absence of IFN agonists or signaling inhibitors. e , Representative flow cytometry plots show p24 + cells after 4 days of infection. f , Frequencies of p24 + cells (of total CD3 + T cells) were quantified per donor under each stimulation condition. Treatment with IFNα, IFNγ, poly I:C or R848 reduced infection, whereas blockade of IFN signaling (using a combination of antibodies against the IFNα/β receptor chain (IFNARi) and IFNγ) increased infection frequency. Comparisons among stimulation conditions (control, IFNα, IFNγ, poly I:C and R848) were analyzed by Friedman’s test followed by Dunn’s post hoc test. All bar and scatter plots display mean ± s.e.m. Unless otherwise indicated, all statistical tests were two-sided, with multiple comparison adjustments applied where specified; otherwise, exact nominal P values are reported in the figures. FMO, fluorescence minus one; Unstim, unstimulated.

Article Snippet: The staining panel included: LIVE/DEAD BV510, Life Technologies, L34957 ; CD3 BUV615, BD Biosciences, 612992, clone UCHT1; CD4 BV605, BioLegend, 317438, clone OKT4; CD45RA BV650, BioLegend, 304136, clone HI100; CD27 AF647, BD Biosciences, 567026, clone O323; CCR7 BUV563, BD Biosciences, 741317, clone 3D12; pIRF3 PerCP, Novus Biologicals, BS-3195R-PERCP, clone polyclonal; pIRF7 BUV496, BD Bioscience, custom, clone K47-671; pSTAT1 (p701) AF488, BD Biosciences, AB_2737715, clone 4a; IFIT1 APC, Novus Biologicals, NBP2-71005APC, clone OTI3G8; APOBEC3G AF700, Novus Biologicals, NBP1-77206AF700, clone polyclonal; and p24 RD1, Beckman Coulter, 6604667, clone KC57.

Techniques: Expressing, Gene Expression, In Vitro, Cell Culture, Fluorescence, Flow Cytometry, Infection, Control, Comparison