Journal: Microbiology Spectrum

Article Title: Attenuation of Avian Flavivirus by Rewiring the Leucine and Serine Codons of Its E-NS1 Protein toward Stop Mutation To Redirect Virus Evolution

doi: 10.1128/spectrum.02921-22

Figure Lengend Snippet: Immunogenicity characteristics of CQW1-OTS in 25-day-old ducks. (A) Survival rate of duck postinfection. The statistical significance of survival was analyzed using a survival curve and the log-rank (Mantel-Cox) test, with significance defined. Ns, not significant. n = 5. (B) Viral copies (at 3 and 5 dpi) in the heart, liver, spleen, lung, kidney, brain, and thymus of 25-day-old ducks were detected by RT-qPCR. (C) Relative mRNA expression of IFN-α, IFN-β, and IL-1β in duck spleen at 5 dpi. (D) Peripheral T-lymphocyte (from infected ducks) proliferation activated by TMUV infection. (E) Serum (14 dpi) levels of IFN-γ and IL-4 were determined using ELISA. (F) Neutralization antibody levels in the serum were determined with PRNT. Statistical significance was determined using an unpaired t test. Ns, not significant; *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

Article Snippet: Lymphocytes were separated from blood using a peripheral blood lymphocyte separation kit (Solarbio, Beijing, China) following the manufacturer’s recommendations.

Techniques: Immunopeptidomics, Quantitative RT-PCR, Expressing, Infection, Enzyme-linked Immunosorbent Assay, Neutralization

Journal: Clinical Cancer Research

Article Title: Integrating Multisector Molecular Characterization into Personalized Peptide Vaccine Design for Patients with Newly Diagnosed Glioblastoma

doi: 10.1158/1078-0432.CCR-23-3077

Figure Lengend Snippet: T-cell characterization. A–C, immunoSEQ Analyzer differential abundance analysis displayed in pairwise scatter plots are shown for subjects 1, 2, and 3, respectively. The frequency of productive TCRβ rearrangements (clones) from the peripheral blood before and after NeoVax is compared. No data are available for subject 4 because they did not receive NeoVax. D–M, Bar graphs displaying IFNγ spots from IFNγ ELISPOT assays. Experiments were performed with duplicate or triplicate wells, and biological replicates were performed at least twice. *, P < 0.05 (Student t test) between pre- and post-NeoVax responses. There were 84, 77, and 35 days between pre- and post-NeoVax peripheral blood collections for subjects 1, 2, and 3, respectively. D, Direct ex vivo stimulation on select CD8 + T cells with minimal epitope peptide pools (m) or DMSO control for subject 1 post-NeoVax. E, Direct ex vivo stimulation on select CD4 + T cells with SLPs or DMSO control for subject 1 post-NeoVax. F, Direct ex vivo stimulation on select CD8 + T cells with minimal epitope peptide pools (m) or DMSO control for subject 1 pre-NeoVax. G, Direct ex vivo stimulation on select CD4 + T cells with SLPs or DMSO control for subject 1 pre-NeoVax. H–M, In vitro expanded PBMCs from pre- and postvaccination time points stimulated with IL2 and corresponding SLPs (green bars), minimal epitope peptide pools (dark orange bars), or DMSO control (purple bars) for 12 days prior to restimulation and analysis for subjects 1, 2, and 3.

Article Snippet: On day 13, cells were restimulated with 1 × 10 5 autologous T-cell depleted PBMCs (STEMCELL Technologies) and respective neoantigen minimal epitope peptide pools, the full-length peptide, or DMSO in a human IFNγ ELISPOT plate (ImmunoSpot).

Techniques: Clone Assay, Enzyme-linked Immunospot, Ex Vivo, Control, In Vitro

Journal: iScience

Article Title: CBP/p300 lysine acetyltransferases inhibit HIV-1 expression in latently infected T cells

doi: 10.1016/j.isci.2024.111244

Figure Lengend Snippet: CBP/p300 inhibition induces HIV-1 expression in CD4 + cells ex vivo (A) Experimental design for RGH infection of primary CD4 + T cells. CD4 + T cells isolated from an uninfected participant were infected with the RGH dual reporter virus at a low MOI. 3 days, post-infection, cells were incubated with a vehicle control (DMSO), 10 μM A-485, or 10 μM iP300w for 24 h after which proviral expression was assessed by flow cytometry. (B) Representative flow cytometry scatterplots of primary CD4 + T cells treated as in (A). Latent infections are in Q1 (mCherry+), productively infected T cells are in Q2 (GFP+/mCherry+), noise generated by viral recombination is in Q3 (GFP+), and uninfected cells are in Q4. (C and D) Following treatment of primary CD4 + T cells as in (A), proviral transcription was determined by flow cytometry and is expressed as the percentage of productive infections (C) and the GFP mean fluorescence intensity (MFI) of infected cells (D) ( n = 3, mean ± SD, one-way ANOVA with Dunnett’s multiple comparisons test). (E) Viability was determined for participant derived CD4 + T cells treated as in (A) ( n = 3, mean ± SD, one-way ANOVA with Dunnett’s multiple comparisons test). (F and G) Primary CD4 + PBMCs isolated from people living with HIV-1 who are receiving ART were treated with a vehicle (DMSO), 10 μM A-485, or 10 μM iP300w and were either left unstimulated or were treated with anti-CD3/anti-CD28. Following 24 h, intracellular RNA was extracted, and RT-PCR was preformed using oligos specific for multiply spliced Tat-Rev HIV-1 mRNA transcripts. HIV-1 mRNA expression is normalized to GAPDH ( n = 3, mean ± SD, unpaired t test). (H) Primary CD4 + PBMCs treated as in (F) and (G) were assessed for cellular viability ( n = 2, mean, unpaired t test). Statistical significance is indicated at ∗ p < 0.05, ∗∗ p < 0.01, or ∗∗∗ p < 0.001, with n.s. denoting non-significant p ≥ 0.05.

Article Snippet: Human Peripheral Blood CD4 + T-cells , StemCell Technologies , 200–0165.

Techniques: Inhibition, Expressing, Ex Vivo, Infection, Isolation, Virus, Incubation, Control, Flow Cytometry, Generated, Fluorescence, Derivative Assay, Reverse Transcription Polymerase Chain Reaction

Journal: iScience

Article Title: CBP/p300 lysine acetyltransferases inhibit HIV-1 expression in latently infected T cells

doi: 10.1016/j.isci.2024.111244

Figure Lengend Snippet:

Article Snippet: Human Peripheral Blood CD4 + T-cells , StemCell Technologies , 200–0165.

Techniques: Virus, Recombinant, SYBR Green Assay, Activation Assay