Journal: PLoS ONE

Article Title: Evaluation of Cellular Phenotypes Implicated in Immunopathogenesis and Monitoring Immune Reconstitution Inflammatory Syndrome in HIV/Leprosy Cases

doi: 10.1371/journal.pone.0028735

Figure Lengend Snippet: Median and interquartile range (IQR) of T cell immunophenotyping percentage values or absolute counts obtained from HIV/leprosy individuals according to viral load (VL) values and from HIV-monoinfected, leprosy patients and healthy controls.

Article Snippet: CD45RA- fluorescein isothiocyanate (FITC, clone HI100, IgG2b) and CD45RO-phycoerythrin (PE, clone UCHL1, IgG2a) (BD-Biosciences, Franklin Lakes, NJ, USA) were used to define cellular differentiation status and dual-color staining with CD8 (FITC, IgG1, clone MCD8) and CD38 (PE, IgG1, clone T16) or CD3 (FITC, IgG2b, clone UCHT1) and HLA-DR (PE, IgG2a, clone BRA30) (IQ Products, Groningen, The Netherlands) antibodies to analyze cellular activation.

Techniques:

Journal: Molecules

Article Title: Recombinant Lactaptin Induces Immunogenic Cell Death and Creates an Antitumor Vaccination Effect in Vivo with Enhancement by an IDO Inhibitor

doi: 10.3390/molecules25122804

Figure Lengend Snippet: Distribution of immune cell populations in peripheral blood. PBMCs were harvested from whole blood samples, stained with CD45, CD3, and CD19 markers, and analyzed by flow cytometry. The number of positive cells is shown as a percentage of total blood CD45+ lymphocytes. ( a ) B-lymphocytes; ( b ) T-lymphocytes. Healthy mice were used as a negative control; “Control vaccination” mice were vaccinated with live MX-7 cells. Statistical differences between control and experimental groups are indicated by * for p < 0.05; ** for p < 0.01.

Article Snippet: The following antibodies and chemicals were obtained from commercial sources: rabbit ant-h/m/rat CRT (Abcam, ab2907, Cambridge, UK), rabbit ant-h/m HMGB1 (Abcam, EPR3507, Cambridge, UK), goat anti-h/m/r GAPDH (R&D Systems, AF5718, Minneapolis, USA), mouse anti-human b-Tubulin (Sigma-Aldrich, T8328, St. Louis, MO, USA), mouse ant-h/rat HSP70 (R&D Systems, 841680, Minneapolis, USA), Alexa Fluor 594-conjugated chicken anti-rabbit antibodies (Invitrogen, Carlsbad, CA, USA), Alexa Fluor 555-conjugated goat anti-mouse antibodies (Invitrogen, Carlsbad, CA, USA), polyclonal rabbit-anti-mouse and mouse-anti-rabbit HRP-conjugated antibodies (Biosan, Novosibirsk, Russia), anti-mouse CD45 PerCP/Cy5.5 (Sony Biotechnologies, 1338530, San Jose, CA, USA), anti-mouse CD19 APC (Sony Biotechnologies, 1177650, San Jose, CA, USA), anti-mouse CD3 FITC (Sony Biotechnologies, 1101015, San Jose, CA, USA), anti-mouse CD25 PE (Sony Biotechnologies, 1110040, San Jose, CA, USA), and anti-mouse CD8a PE (R&D Systems, FAB116P, Minneapolis, USA).

Techniques: Staining, Flow Cytometry, Negative Control, Control

Journal: Nature Communications

Article Title: Tumor response and endogenous immune reactivity after administration of HER2 CAR T cells in a child with metastatic rhabdomyosarcoma

doi: 10.1038/s41467-020-17175-8

Figure Lengend Snippet: a Histological examination of the bone marrow 4 weeks after the salvage chemotherapy (ARST0921) and prior to initiating CAR T-cell infusions showing hypocellularity and presence of rhabdomyosarcoma (RMS) cells on hematoxylin and eosin (H&E) staining and immunoreactivity to desmin and myogenin, b complete disease response (CR1), evidenced by recovery of trilineage hematopoiesis and absence of immunoreactivity to desmin and myogenin after three HER2 CAR T-cell infusions. Panels ( a , b ) show representative microscopic images; scale bar 100 µm. c Representative image from positron emission tomography–computed tomography (PET-CT) with no evidence of FDG-avid disease in bone marrow or other sites 6 weeks after the third HER2 CAR T-cell infusion. d Detection of HER2 CAR-expressing T cells in the peripheral blood 7 days after the second infusion using flow cytometry. HER2 CAR was specifically recognized using HER2.Fc chimeric protein followed by a goat anti-human Fc conjugated with PE as a secondary antibody. SSC side scatter. e The proportion of CD3+ HER2 CAR-expressing T cells on day +7 after each infusion during the induction period. f Histograms showing the PD-1 and LAG3 surface expression in CAR-positive CD8+ (in blue) in comparison to CAR-negative CD8+ T cells (in black) at peak expansion (day +7) after each infusion during induction, and g the corresponding median fluorescence intensity (MFI) of PD-1 and LAG3 surface expression in CAR-positive and CAR-negative CD8+ T cells.

Article Snippet: The following monoclonal antibodies were used: mouse anti-human CD3 FITC (stock solution, 5 μL/100 μL test at 1:20 dilution, clone: SK7, Cat# 349201, Lot# 7221481, Becton Dickinson Biosciences, San Jose, CA), mouse anti-human CD8 Pacific Blue (stock solution, 5 μL/100 μL test at 1:20 dilution, clone: B9.11, Cat# A82791, Lot# 38, Beckman Coulter, Indianapolis, IN), mouse anti-human CD279 (PD-1) APC (stock solution, 5 μL/100 μL test at 1:20 dilution, clone: MIH4, Cat# 558694, Lot# 8012764, Becton Dickinson Biosciences, San Jose, CA), mouse anti-human CD223 (LAG3) FITC (stock solution, 5 μL/100 μL test at 1:20 dilution, clone: 3DS223H, Cat# 11-2239-42, Lot# 4337363, Thermo Fisher Scientific, Waltham, MA).

Techniques: Staining, Positron Emission Tomography, Computed Tomography, Positron Emission Tomography-Computed Tomography, Expressing, Flow Cytometry, Comparison, Fluorescence