Journal: iScience

Article Title: Chemical and biological characterization of vaccine adjuvant QS-21 produced via plant cell culture

doi: 10.1016/j.isci.2024.109006

Figure Lengend Snippet: In vitro biological characterization; ccQS-21 and beQS-21 equivalently induce hemolysis, trigger the innate immune cell stress response, and enhance antigen processing and presentation to enhance the potency of the vaccinal immune synapse (A) Upon incubation with human red blood cells both beQS-21 and ccQS-21 retain equivalent saponin-specific hemolytic properties at pH-7.4 with matching half maximal effective concentration (EC50). (B) Upon pre-conditioning of THP1 human myeloid cells with a TLR4 agonist (LPS), both beQS-21 and ccQS-21 induce inflammasome-dependent secretion of IL-1β and the production of alarmins associated with pyroptosis such as HMGB1. Both IL-1β secretion and pyroptosis can be regulated by the canonical inflammasome pathway and can be suppressed by the disruption of the genes encoding for NLRP3, ASC and Caspase-1 or by the use of Caspase-1 inhibitor, ZVAD. (C) Both beQS-21 and ccQS-21 can comparably induce a broader inflammatory response in vitro . The breadth of the inflammatory response induced by the stimulation of human granulocyte macrophage colony stimulating factor (GMCSF) and IL-4 differentiated Monocyte derived Dendritic Cells (Mo-DCs) using both QS-21 was assessed with multiplex cytokine quantification. Both beQS-21 (BE) and ccQS-21 (CC) induce the same inflammatory signature which is further enhanced by formulating with TLR4 agonist. (D) Pseudo cross-presentation assay using cell viability to demonstrate that beQS-21 and ccQS-21 equivalently induce cytotoxicity through the regulation of the cytosolic translocation from the phagolysosome (e.g., as of the indicated toxin: saporin), a critical process for antigen cross presentation. (E) Flow cytometry analysis demonstrating that both QS-21 equivalently enhance large antigen processing and cross-presentation on MHC-I using a T cell receptor (TCR)-like antibody against the H-2K b -SIINFEKL complex. (F) In vitro antigen cross-presentation assay demonstrating that ccQS-21 and beQS-21 equivalently enhance antigen processing and minimal epitope cross-presentation to activate transgenic T cells with matched OT1 TCR to express NFAT luciferase reporter. (G) OT1 T cell proliferation shows ccQS-21 and beQS-21 equivalently enhance antigen cross-presentation to induce the proliferation of primary T cells with matched OT1 TCR. OVA: Ovalbumin, OT1 long peptide: 29mer peptide including SIINFEKL, BFM: Bafilomycin A1, inhibiting the vacuolar type H + -ATPase (v-ATPase) to transfer protons into the lysosome.

Article Snippet: Cells were then plated in flat-bottom culture 96-well plates (0.5 × 10 6 cells per well). gE overlapping peptide mix (0.25 μg/mL, JPT Peptide Technologies, Germany), or OT1 and OT-II peptide mix (5 μg/mL, GenScript, China) were added for restimulation.

Techniques: In Vitro, Incubation, Concentration Assay, Disruption, Derivative Assay, Multiplex Assay, Translocation Assay, Flow Cytometry, Transgenic Assay, Luciferase

Journal: iScience

Article Title: Chemical and biological characterization of vaccine adjuvant QS-21 produced via plant cell culture

doi: 10.1016/j.isci.2024.109006

Figure Lengend Snippet:

Article Snippet: Cells were then plated in flat-bottom culture 96-well plates (0.5 × 10 6 cells per well). gE overlapping peptide mix (0.25 μg/mL, JPT Peptide Technologies, Germany), or OT1 and OT-II peptide mix (5 μg/mL, GenScript, China) were added for restimulation.

Techniques: Recombinant, Cell Isolation, Enzyme-linked Immunospot, Cytotoxicity Assay, Luciferase, Expressing

Journal: Frontiers in Oncology

Article Title: Biological Predictors of De Novo Tumors in Solid Organ Transplanted Patients During Oncological Surveillance: Potential Role of Circulating TERT mRNA

doi: 10.3389/fonc.2021.772348

Figure Lengend Snippet: Biological parameters in the 49 SOT patients after stratification by tumor occurrence.

Article Snippet: CMV, EBV, Survivin, TERT peptide mixes (ProImmune, Oxford UK; 0.2ng/mL of each peptide mix) or unspecific stimuli (0.5mg/mL αCD3/αCD28) were resuspended in CTL-test Medium, plated in triplicate and incubated for 10-20 minutes.

Techniques:

Journal: Frontiers in Oncology

Article Title: Biological Predictors of De Novo Tumors in Solid Organ Transplanted Patients During Oncological Surveillance: Potential Role of Circulating TERT mRNA

doi: 10.3389/fonc.2021.772348

Figure Lengend Snippet: Cox regression analysis evaluating the associations between baseline demographic, clinical or biological parameters and tumor onset.

Article Snippet: CMV, EBV, Survivin, TERT peptide mixes (ProImmune, Oxford UK; 0.2ng/mL of each peptide mix) or unspecific stimuli (0.5mg/mL αCD3/αCD28) were resuspended in CTL-test Medium, plated in triplicate and incubated for 10-20 minutes.

Techniques:

Journal: Scientific Data

Article Title: Quantitative Mass Spectrometry Analysis of Cerebrospinal Fluid Protein Biomarkers in Alzheimer’s Disease

doi: 10.1038/s41597-023-02158-3

Figure Lengend Snippet: Isotopologue peptide internal reference standards to determine consistency of LC-MS/MS platform. Each of the CSF samples were spiked with a six-peptide, 5 isotopologue concentration LC-MS/MS Peptide Reference Mix from Promega (50 fmol/µL). ( a ) Extracted ion chromatogram for the 6 peptide (1pmol) mixture illustrating the wide range of retention times due to their hydrophobicity. ( b ) The raw peak areas in 423 injections over 5 days were used to determine the label-free CV for each peptide isotopologue estimating the lowest limits of detection to be between 1–10 fmole for each peptide. ( c ) The 5 unique isotopologues are used to assess the dynamic range across the gradient profile and each peptide demonstrates linearity across 3–4 orders of magnitude in the batch of 423 injections. Error bars represent the standard deviation across 423 injections.

Article Snippet: Each of the CSF samples were spiked with a six-peptide, 5 isotopologue concentration LC-MS/MS Peptide Reference Mix from Promega (50 fmol/μL). ( a ) Extracted ion chromatogram for the 6 peptide (1pmol) mixture illustrating the wide range of retention times due to their hydrophobicity. ( b ) The raw peak areas in 423 injections over 5 days were used to determine the label-free CV for each peptide isotopologue estimating the lowest limits of detection to be between 1–10 fmole for each peptide. ( c ) The 5 unique isotopologues are used to assess the dynamic range across the gradient profile and each peptide demonstrates linearity across 3–4 orders of magnitude in the batch of 423 injections.

Techniques: Liquid Chromatography with Mass Spectroscopy, Concentration Assay, Standard Deviation