lymphoprep cell gradient centrifugation  (STEMCELL Technologies Inc)

Journal: The Journal of Infectious Diseases

Article Title: Enhanced Human Immunodeficiency Virus-1 Replication in CD4 + T Cells Derived From Individuals With Latent Mycobacterium tuberculosis Infection

doi: 10.1093/infdis/jiaa257

Figure Lengend Snippet: Mycobacterium tuberculosis infection leaves an immunological footprint detected by elevated levels of inflammation-associated analytes in plasma and that is dependent on the infection status. (A) Experimental outline. Whole blood was collected from volunteers representing the subject groups shown in Table 1. A portion of blood was kept separate for harvesting plasma to be used for multiplex cytokine analysis and for supplementing the culture media to be used for CD4+ T cells. The rest of the blood was used for isolating peripheral blood mononuclear cells (PBMC) for flow cytometry analysis and for isolating CD4+ T cells for human immunodeficiency virus (HIV)-1 infection experiments. Human immunodeficiency virus-1 infection was monitored by enzyme-linked immunosorbent assay (ELISA) of HIV-1 antigen (p24), quantitative polymerase chain reaction (qPCR) analysis of HIV-1 deoxyribonucleic acid, and qPCR analysis of HIV-1 ribonucleic acid. (B) Plasma analyte concentrations were measured by multiplex analysis and are displayed as a heatmap for each blood donor. Heatmap shows the amount of each analyte per blood donor, relative to the maximum signal for set analyte. (C) Plots of representative analytes indicating data range, median, and interquartile range. Each point represents individual donors. Dotted red line indicates the limit of detection. P values were calculated based on the Mann-Whitney U test: *, P < .05 and **, P < .01. ‡ denotes P < .0019 after Bonferroni correction. The full data set of analytes is included as Supplementary Table and Supplementary Figure 1A–C. FACS, fluorescence-activated cell sorting; IFN, interferon; IL, interleukin; LTBI, latent tuberculosis infection; TB, tuberculosis.

Article Snippet: Peripheral Blood Mononuclear Cells Isolation and CD4 + T-Cell Purification Peripheral blood mononuclear cells (PBMC) were isolated using the Lymphoprep density gradient (StemCell Technologies).

Techniques: Infection, Clinical Proteomics, Multiplex Assay, Flow Cytometry, Virus, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Fluorescence, FACS

Journal: The Journal of Infectious Diseases

Article Title: Enhanced Human Immunodeficiency Virus-1 Replication in CD4 + T Cells Derived From Individuals With Latent Mycobacterium tuberculosis Infection

doi: 10.1093/infdis/jiaa257

Figure Lengend Snippet: Mycobacterium tuberculosis infection does not affect the overall proportion of activated CD4+ T cells in the blood. Peripheral blood mononuclear cells from study subjects were stained for flow cytometry analysis to determine the proportion of CD4+ T cells expressing any of the molecules associated with T-cell activation: CD25, CD69, and/or HLA-DR. This combination of markers captures both early and late T-cell activation states. Representative fluorescence-activated cell sorting plots for each subject group and a plot of all donors are shown. Plots indicate data range, median, and interquartile range. P values were calculated based on the Mann-Whitney U test. FSC, forward scatter; LTBI, latent tuberculosis infection; TB, tuberculosis.

Article Snippet: Peripheral Blood Mononuclear Cells Isolation and CD4 + T-Cell Purification Peripheral blood mononuclear cells (PBMC) were isolated using the Lymphoprep density gradient (StemCell Technologies).

Techniques: Infection, Staining, Flow Cytometry, Expressing, Activation Assay, Fluorescence, FACS, MANN-WHITNEY

Journal: The Journal of Infectious Diseases

Article Title: Enhanced Human Immunodeficiency Virus-1 Replication in CD4 + T Cells Derived From Individuals With Latent Mycobacterium tuberculosis Infection

doi: 10.1093/infdis/jiaa257

Figure Lengend Snippet: Mycobacterium tuberculosis infection leaves an immunological footprint detected by changes in the relative proportion of CD4+ T-cell memory subsets. Peripheral blood mononuclear cells from study subjects were stained for flow cytometry analysis to determine the proportion of CD4+ T-cell subsets based on the expression of molecules associated with memory phenotypes. A representative dot plot is shown with the gating strategy and the CD4+ T-cell subsets represented by each gate: effector memory (TEM), central memory (TCM), and terminally differentiated (TTD). CCR7+/CD45RA+ cells were further subdivided into true naive cells (TN) and stem cell memory T cells (TSCM) based on the expression of CD27 and CD95. Percentages shown for each gate indicate their proportion relative to the total CD4+ T-cell population. Plots represent the proportion of each CD4+ T-cell subset for all donors and show the data range, median, and interquartile range. P values were calculated based on the Mann-Whitney U test: *, P < .05 and **, P < .01. LTBI, latent tuberculosis infection; TB, tuberculosis.

Article Snippet: Peripheral Blood Mononuclear Cells Isolation and CD4 + T-Cell Purification Peripheral blood mononuclear cells (PBMC) were isolated using the Lymphoprep density gradient (StemCell Technologies).

Techniques: Infection, Staining, Flow Cytometry, Expressing, MANN-WHITNEY