Journal: Clinical and Experimental Immunology

Article Title: The effect of human immunodeficiency virus-1 on monocyte-derived dendritic cell maturation and function

doi: 10.1111/j.1365-2249.2012.04628.x

Figure Lengend Snippet: Human immunodeficiency virus (HIV)-1 infection of monocyte-derived dendritic cells (MDDC) inhibits antigen presentation to autologous CD8+ T cells. HIV-1CS204-infected immature MDDC (iMDDC) were pulsed with a cytomegalovirus, Epstein–Barr and influenza viruses (CEF) peptide pool and then co-cultured with autologous 5 mM CellTrace™ carboxyfluorescein succinimidyl ester (CFSE)-stained peripheral blood mononuclear cells (PBMC) at a ratio of 1:10 for 7 days. While preincubation of uninfected iMDDC induced the proliferation of autologous monocyte depleted PBMC, HIV-1CS204 infection of iMDDCs impaired the presentation of a CEF peptide pool as measured by CFSE dilution. Results are expressed as means ± standard deviation. *P = 0·02 by paired t-test (n = 6).

Article Snippet: MDDCs, differentiated and infected as above, were pulsed for 3 h with 3 μg/ml of a CEF peptide pool containing 23 human leucocyte antigen (HLA)-ABC-restricted T cell epitopes from human cytomegalovirus, Epstein–Barr and influenza viruses (CEF) (Anaspec Inc., Fremont, CA, USA).

Techniques: Virus, Infection, Derivative Assay, Immunopeptidomics, Cell Culture, Staining, Standard Deviation

Journal: Frontiers in Immunology

Article Title: A SARS-CoV-2 NSP7 homolog of a Treg epitope suppresses CD4+ and CD8+ T cell memory responses

doi: 10.3389/fimmu.2023.1290688

Figure Lengend Snippet: Effect of SARS-CoV-2-NSP7 peptide on CD8+ T cell response in PBMCs from healthy donors stimulated with CEF peptides. PBMCs from healthy donors were stimulated with 0.5 µg/ml of CEF alone or in combination with FV621 or increasing concentrations of 289, 289z or SARS-CoV-2 NSP7 peptide (10, 20 and 40μg/ml). Proliferation of CD8+ T cells were measured six days post-stimulation by flow cytometry based on CFSE dilution. (A) Left panel shows the gating of CD8+ T cells. In the right panel, representative flow plots display the dose effect on the proliferation of CD8+ T cells resulting from increasing concentrations of SARS-CoV-2-NSP7-homolog peptide. (B, C) Graphs show the (%) proliferation of CD8+ T cells for each peptide tested in the assay, compared against CEF stimulation alone so that percent inhibition can be calculated or inferred. FV621 was used as a positive control as it has demonstrated consistent suppression of CEF response across donors in our previous publication . Original data is presented in panel B, and data normalized for CEF stimulation as 100% proliferation in panel C. The percent inhibition of CD8+ T cell proliferation for each peptide at each concentration tested in the assay compared to CEF stimulation alone. Data are combined from five donors in the experiment. P values * = <0.05, ** = <0.01 and *** = <0.0002 represent statistical significance vs . CEF stimulation alone using a two-tailed t test.

Article Snippet: To determine the effects of NSP7-289 on CD8+ T cell recall responses, and demonstrate that both CD4+ and CD8+ T cells can be suppressed by the Treg epitopes, PBMCs from healthy donors were stimulated with 0.5 μg/ml of a mixture of MHC class I peptides derived from cytomegalovirus, Epstein-Barr virus, and influenza virus designated CEF (CEF peptide pool, Mabtech, cat no. 3615-1), alone or with the of test peptides.

Techniques: Flow Cytometry, Inhibition, Positive Control, Concentration Assay, Two Tailed Test