Journal: PloS one

Article Title: Menstrual blood as a potential source of endometrial derived CD3+ T cells.

doi: 10.1371/journal.pone.0028894

Figure Lengend Snippet: Figure 6. CMV-specific T cells derived from the menstrual blood. A. PBMC B. MBC from the same healthy woman (#1). Cells were stimulated with overlapping peptides from CMV pp65 and PMA/I as a positive control. Both IFN-c and TNF-a secretion are shown. Percent of cytokine positive cells (IFN-c or TNF-a) are shown above the gate. doi:10.1371/journal.pone.0028894.g006

Article Snippet: CMV-pp65 peptide pool (69 18-mers, overlapping by 10 spanning the entire amino acid sequence of the HCMV pp65 protein, New England Peptide, Gardner, MA) (2 mg/ml) or CMV-Lysate (Advance Biotechnologies, Columbia, MD) (2 mg/ ml) was used to stimulate T cells.

Techniques: Derivative Assay, Positive Control

Journal: PloS one

Article Title: Menstrual blood as a potential source of endometrial derived CD3+ T cells.

doi: 10.1371/journal.pone.0028894

Figure Lengend Snippet: Figure 7. Reduced frequency of CMV-specific T cells derived from the menstrual blood. PBMC and MBC from the same women were stimulated with overlapping peptides from CMV pp65 or CMV lysate and stained for IFN-c. Individual symbols represent paired samples from the same individual. Open symbols represent CD8 T cell responses, closed symbols represent CD4 T cell responses and gray symbols are responses from the healthy volunteer. Statistical compar- isons were made using Wilcoxon Signed Rank test. doi:10.1371/journal.pone.0028894.g007

Article Snippet: CMV-pp65 peptide pool (69 18-mers, overlapping by 10 spanning the entire amino acid sequence of the HCMV pp65 protein, New England Peptide, Gardner, MA) (2 mg/ml) or CMV-Lysate (Advance Biotechnologies, Columbia, MD) (2 mg/ ml) was used to stimulate T cells.

Techniques: Derivative Assay, Staining

Journal: Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico

Article Title: Evaluation of safety and efficacy of p53MVA vaccine combined with pembrolizumab in patients with advanced solid cancers

doi: 10.1007/s12094-018-1932-2

Figure Lengend Snippet: p53MVA/pembrolizumab activate p53-specific T cell responses and associated immune function pathways. The response of CD8+ and CD4+ T cells from PBMCs after 24-h stimulation culture with p53MVA, MVA, p5396, and pp65138, as determined by flow cytometric analysis, is shown for five patients in columns a and b. The upregulation of CD137 expression on the surface of T cells in response to specific recall stimuli reflects increased frequencies of p53-reactive T cells in the circulation after vaccination. Culture conditions: NIL – medium alone; p53(96) – pool of peptides derived from wild type p53 sequence; pp65(138) – control peptides derived from pp65 CMV; MVA – wild type MVA vaccinia virus; p53MVA – recombinant MVA virus. Column c shows CD4+/CD8+ T cell ratio data for patients UPN003, UPN008, and UPN006 with declining ratio who benefited from the treatment and two patients UPN002 and UPN004 (and all other patients in Fig. 2) whose ratio remained stable and unaffected by the treatment. Column d summarizes data from multiplexed gene expression analysis of PBMC samples from indicated patients using nCounter PanCancer Immune Profiling Panel. The analysis of 730 immune profiling genes included selected genes that define immune function pathways. The pathway scores are plotted to show how they vary across time during treatment. The T cell functions and associated immune response categories remained at elevated levels for prolonged period of time in 2/3 patients responding to the treatment. Data presented for UPN003 in columns a, b, and d have been modified with additional time points that were not available at the time of their original publication in Yuan et al. [15].

Article Snippet: In the initial analysis, PBMC were thawed and plated at 2 x 10 5 cells/0.2 ml/well in media (RPMI, FBS 10%, glutamine 2 mM, sodium pyruvate 1 mM, non-essential amino acids) with one of the following stimuli: media alone (NIL), p53MVA, MVA, pool of 96 15-mer overlapping peptides spanning the entire length of p53 (p53 96 ; 5 μg/ml; synthesized in-house), and a control pool of 138 peptides derived from CMV pp65 protein epitopes (pp65 138 ; 2 μg/ml; BEI Resources, NIH, Bethesda).

Techniques: Expressing, Derivative Assay, Sequencing, Control, Virus, Recombinant, Gene Expression, Modification