Journal: Retrovirology

Article Title: Opposing regulation of endolysosomal pathways by long-acting nanoformulated antiretroviral therapy and HIV-1 in human macrophages

doi: 10.1186/s12977-014-0133-5

Figure Lengend Snippet: NanoATV regulation of cytokine profiles in HIV-1 infected MDM. MDM were treated with 100 μM native ATV or nanoATV and infected with HIV-1 at day 0, 5 or 10 post-drug treatment. Untreated, uninfected cells were used as controls. After 24 hours of viral infection, cell culture media were collected and analyzed using a cytokine bead array. (A) Density plots show expression of IL-12, TNF, IL-10, IL-6, IL-1β and IL-8 for control, treated, HIV-1 infected and treated infected MDM. Cytokine levels were detected by FACSArray cytometer and data was plotted using FlowJo (version 10.7) software. (B) Levels of IL-12 and TNF after treatment and infection are shown. Data are analyzed using FCAP software and values of cytokine expression were expressed as pg/mL.

Article Snippet: 24 hours after drug treatment, 50 μL cell culture media from treated and infected MDM were tested to determine the concentrations of inflammatory cytokines measured by a cytokine bead array (CBA) detection kit (Becton Dickinson Biosciences; Mississauga, ON, USA) and performed according to instructions of the manufacturer.

Techniques: Infection, Cell Culture, Expressing, Control, Cytometry, Software

Journal: The Journal of Biological Chemistry

Article Title: Human Memory, but Not Naive, CD4 + T Cells Expressing Transcription Factor T-bet Might Drive Rapid Cytokine Production *

doi: 10.1074/jbc.M114.608745

Figure Lengend Snippet: Kinetics of cytokine secretion by activated naive and memory CD4+ T cells. Naive and memory CD4+ T cells were isolated from CD4+ T cells by MACS microbeads, and the purity of naive or memory CD4+ T cells was more than 97% as determined by flow cytometry (a). Naive and memory CD4+ T cells were stimulated with (open histogram) or without (shaded histogram) PMA and ionomycin in the presence of BFA for 6 h. Cells were stained by intracellular staining and analyzed by FACS (b). Purified naive and memory CD4+ T cells were stimulated with immobilized anti-CD3 (1 μg/ml) and anti-CD28 (1 μg/ml) for 0 to 72 h, and the concentrations of cytokines IFN-γ, IL-2, and TNF-α were detected by ELISA (c). Data are representative of three separate experiments with similar results.

Article Snippet: Human ELISA kits for cytokines IFN-γ, IL-2, and TNF-α were purchased from BD Biosciences.

Techniques: Isolation, Flow Cytometry, Staining, Purification, Enzyme-linked Immunosorbent Assay

Journal: BioMed Research International

Article Title: A De- O -acylated Lipooligosaccharide-Based Adjuvant System Promotes Antibody and Th1-Type Immune Responses to H1N1 Pandemic Influenza Vaccine in Mice

doi: 10.1155/2016/3713656

Figure Lengend Snippet: Th1-type-predominant immune response to CIA06-adjuvanted influenza vaccine. Splenocytes were isolated from the mice that had been immunized as described in and cultured for 72 h in the absence (■) or presence of Greenflu-S (■). The levels of IFN- γ (a) and IL-5 (b) in the culture media were determined by sandwich ELISA, and IFN- γ : IL-5 ratios were calculated for each group (c). Statistical differences were analyzed by one-way ANOVA followed by Tukey's multiple comparison test. Results are expressed as the means ± SD of values obtained from triplicate cultures that used two spleens each. Data shown are representative of three independent experiments. ∗ P < 0.05; ∗∗ P < 0.01; and ∗∗∗ P < 0.001. NS, not significant.

Article Snippet: Recombinant mouse IL-2 was acquired from R&D Systems (Minneapolis, MN, USA), and IFN- γ and IL-5 cytokine ELISA kits were obtained from BD Biosciences (San Diego, CA, USA) and R&D Systems.

Techniques: Isolation, Cell Culture, Sandwich ELISA, Comparison

Journal: BioMed Research International

Article Title: A De- O -acylated Lipooligosaccharide-Based Adjuvant System Promotes Antibody and Th1-Type Immune Responses to H1N1 Pandemic Influenza Vaccine in Mice

doi: 10.1155/2016/3713656

Figure Lengend Snippet: CIA06-adjuvanted influenza vaccine stimulates both CD4 + and CD8 + T cell responses. Splenocytes were isolated from the mice ( n = 6) that had been immunized twice at a 3-week interval with nonadjuvanted or adjuvanted Greenflu-S (0.2 μ g) and stimulated with the vaccine for 72 h in the absence or presence of anti-CD4 and/or anti-CD8 antibodies. IFN- γ levels in the culture media were determined by sandwich ELISA. Statistical differences were analyzed by one-way ANOVA followed by Tukey's multiple comparison test. Results are expressed as the means ± SD of values obtained from triplicate cultures that used two spleens each. Data shown are representative of three independent experiments. ∗ P < 0.05; ∗∗ P < 0.01; and ∗∗∗ P < 0.001. NS, not significant.

Article Snippet: Recombinant mouse IL-2 was acquired from R&D Systems (Minneapolis, MN, USA), and IFN- γ and IL-5 cytokine ELISA kits were obtained from BD Biosciences (San Diego, CA, USA) and R&D Systems.

Techniques: Isolation, Sandwich ELISA, Comparison

Journal: Oncotarget

Article Title: Roles of calpain-calpastatin system (CCS) in human T cell activation

doi: 10.18632/oncotarget.13259

Figure Lengend Snippet: Concentrations of various Th1, Th2, Th17 and proinflammatory cytokines (indicated in respective panels) were estimated using the CBA Flex™ cytometric technique as described in the Materials and Methods. Mean concentrations (bars) and standard deviations (whiskers) are indicated. P values are given in the panels whenever the difference between control and CI-treated cytokine concentration was statistically significant. N.s. - not significant. Student T test with Welch's correction was applied. N = 11 for each cytokine determination.

Article Snippet: Concentrations of various cytokines (IFNγ, IL-1β, IL-2, IL-6, IL-8, IL-10, IL-12p70, IL-17 and TNFα) in the culture supernatants of untreated PBMC or of PBMC treated with 10 μMCI-II or CI-IV were determined cytometrically using the Cytokine Beads Array FlexTM kits (Becton Dickinson, USA) according to the manufacturer's instructions.

Techniques: Control, Concentration Assay