Journal: The EMBO Journal

Article Title: A novel pathway combining calreticulin exposure and ATP secretion in immunogenic cancer cell death

doi: 10.1038/emboj.2011.497

Figure Lengend Snippet: Tumour cells dying under phox-ER stress conditions induce DC maturation and activate the adaptive immune system. (A) In-vitro phagocytosis of T24 cells treated with Hyp-PDT (red) by human immature dendritic cells (hu-iDCs) (green). The confocal fluorescence images show various phagocytic interactions between dying T24 cells and hu-iDCs, such as tethering (a), initiation of engulfment by extending the pseudopodia (b), and final stages of engulfment (c); scale bar=20 μm. (B) Human DC maturation analysis. T24 cells were left untreated (CNTR), freeze/thawed (accidental necrosis=AN), or treated with a high PDT dose. They were then co-incubated with hu-iDCs. As a positive control, hu-iDCs were stimulated with LPS for 24 h. After co-incubation, the cells were immunostained in two separate groups for CD80/CD83 positivity and CD86/HLA-DR positivity and scored by FACS analysis. Data have been normalized to the ‘CNTR T24 + hu-iDCs' values. Fold change values are means of two independent experiments (two replicate determinations in each)±s.e.m. (*P<0.05, versus ‘CNTR T24+hu-iDCs'). (C, D) Cytokine and respiratory burst patterns exhibited by human DCs. The T24-hu-iDC co-incubation conditioned media obtained during the experiments detailed in (B) were recuperated followed by analysis for concentrations of nitrite (solubilized form of nitric oxide or NO) (C), and IL-10 (D). Absolute concentrations are the means of two independent experiments (four replicate determinations in each)±s.d. (*P<0.05 versus hu-iDC only). (E) Priming of adaptive immune system by dead/dying CT26 cells. Following immunization with PBS (CNTR) or with CT26 cells treated with tunicamycin (TUN), mitoxantrone (MTX) and the highest PDT dose, the mice were rechallenged with live CT26 tumour cells. Subsequently, the percentage of mice with tumour-free rechallenge site was determined (n represents the number of mice).

Article Snippet: Immunoreactive levels of human IL-1β (MPXHCYTO-60K-04) and IL-10 (MPXHCYTO-60K-04) were measured in the CCM by using Milliplex human cytokines (Merck Millipore, Billerica, MA, USA).

Techniques: In Vitro, Fluorescence, Incubation, Positive Control

Journal: Viruses

Article Title: Distinguishing Incubation and Acute Disease Stages of Mild-to-Moderate COVID-19

doi: 10.3390/v14020203

Figure Lengend Snippet: Kinetics of plasmablast proportions and SARS-CoV-2 specific immunoglobulin enrichment delineated disease stages during mild-to-moderate COVID-19. ( A ) Representative pseudocolor plots of CD38 and CD27 expression data of CD19 + CD45RA + B lymphocytes showed the disease stage-dependent increase of plasmablast proportions (rectangular gate). ( B ) Top left: Quantitative data of plasmablast abundances at different disease stage. Top right, bottom left and bottom right: Semi-quantitative data of SARS-CoV-2 specific IgM, IgG and IgA titers in plasma samples. The data for anti-RBD, -S1, -S2 and –N were combined for IgM and IgG, respectively.* p < 0.05; ** p < 0.01; *** p < 0.001; Tukey-Kramer test for plasmablasts, Kruskal-Wallis test with Dunn’s correction for multiple comparisons for Antibody isotypes. MFI: Median fluorescence intensity. A 450 : Absorbance at 450 nm.

Article Snippet: For the semi-quantitative analyses of IgM and IgG specific for the nucleocapsid (N) or the spike protein subunits S1, S2 and receptor binding domain (RBD), we used the respective MILLIPLEX SARS-CoV-2 antigen panel kits (Merck) to the manufacturer’s instructions.

Techniques: Expressing, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Obesity and Dyslipidemia Synergistically Exacerbate Psoriatic Skin Inflammation

doi: 10.3390/ijms23084312

Figure Lengend Snippet: Measurement of multiple adipokine levels in the mouse serum: ( a – d ) Serum levels of leptin, resistin, IL-6, and MCP-1. Serum samples were collected from wild-type and Apoe −/− mice fed with a normal diet or a high-fat diet and treated with imiquimod cream (25 mg/mouse) or Vaseline (control) ( n = 6 per group). Serum adipokine levels were measured by multiplex assay. ( e ) Serum IL-17A concentrations determined by ELISA. Values are presented as the mean ± standard deviation; * p < 0.05, † p < 0.05, compared to Vaseline-treated mice. IMQ, imiquimod; WT, wild-type; ND, normal diet; HFD, high-fat diet.

Article Snippet: Adipokine profiles in mouse sera were determined using a Milliplex mouse adipokine magnetic bead panel (Merck Millipore, Billerica, MA, USA), as reported previously [ ].

Techniques: Cream, Control, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation