Journal: Scientific Reports

Article Title: Targeting Neph1 and ZO-1 protein-protein interaction in podocytes prevents podocyte injury and preserves glomerular filtration function

doi: 10.1038/s41598-017-12134-8

Figure Lengend Snippet: ISD attenuates NTS induced injury in mice: ( A ) The schematic presentation of experimental plan is shown. ( B – D ) Representative images of pre-injection, 24 hours and day 5 post NTS injection urine samples from NTS + vehicle and NTS + ISD mice as analyzed by SDS-PAGE and coomassie blue staining (left panel). Quantitative analysis (right panel) shows a significant decrease in urine albumin/creatinine ratio in mice treated with ISD at day 5 (p < 0.05), when compared to the vehicle control. ( E ) Immunofluorescence analysis of kidney sections by DAPI (Blue) and Neph1 (Green) and ZO1 (Red) antibodies shows NTS-induced mislocalization of Neph1 and ZO-1 (NTS + Vehicle), whereas treatment with ISD restored the colocalization of Neph1 and ZO-1 at podocyte cell membrane (NTS + ISD).

Article Snippet: Urine albumin/creatinine ratios were obtained using an enzyme-linked immunosorbent assay (ELISA) Albuwell kit (Exocell) and creatinine companion kit (Exocell), and results were analyzed by an unpaired one-tailed t test (GraphPad Prism 7).

Techniques: Injection, SDS Page, Staining, Control, Immunofluorescence, Membrane

Journal: Cell reports. Medicine

Article Title: GRAd-COV2 vaccine provides potent and durable humoral and cellular immunity to SARS-CoV-2 in randomized placebo-controlled phase 2 trial.

doi: 10.1016/j.xcrm.2023.101084

Figure Lengend Snippet: Figure 4. Spike-specific T cell response after GRAd-COV2 vaccination PBMCs were isolated and cryopreserved for the analysis of T cell responses from a subset of 54 volunteers, 21 enrolled in placebo (PL), 17 in SD, and 16 in RD arms. (A) Total T cell response to SARS-CoV-2 spike at days 22 (post-dose 1-PD1) and 36 (post-dose 2-PD2), evaluated by IFNg ELISpot and expressed as IFNg spot- forming cells (SFC) per million PBMCs. (B) Breadth of response to spike: response to DMSO (negative control: gray symbols) and peptide pools covering the S1a (pink symbols), S1b (green symbols), S2a (purple symbols), and S2b (violet symbols) portions of spike, evaluated in ELISpot at day 22. (C) Cross-reactivity of the T cell response to variants of concern: total spike response to Wuhan and Delta or Wuhan and Omicron variants, evaluated in distinct ELISpot assays using day 22 PBMCs from all GRAd-COV2-vaccinated subjects. The response on each variant in an individual volunteer is connected by a line, and bars are set at GM. (D) IFNg (Th1) and IL-5 (Th2) production upon spike peptide pool stimulation, evaluated at the day 36 visit in eight subjects per vaccine arm and two PL recipients by two-color ELISpot. (E and F) Multiparametric flow cytometry analysis of CD4 and CD8 T cells responses at day 36 in all GRAd-COV2 and eight PL recipients. Total spike response (E) and breadth (F) of response on S1 and S2 spike domains. Data are expressed as the percentage of CD4 and CD8 expressing any combination of the analyzed functions (IFNg, TNF-a, IL-2, or CD107a) within the CD69+ fraction in response to spike antigen stimulation. Pie charts (base: median) representing the functional profile of spike-specific CD4 and CD8 are shown in (E) and are better described in Figure S7. In all panels, red circles represent SD arm subjects, blue circles represent RD arm subjects, gray circles represent PL arm subjects, and black horizontal lines indicate GM.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Brilliant Stain Buffer BD Biosciences Cat# 563794 BD Fixation/Permeabilization Kit BD Biosciences Cat# 554714 FVS780 BD Biosciences Cat# 565388 HyClone Defined FBS, US Origin, 500 mL Cytiva Cat# SH30070.03 CTL-WashTM Supplement 10x Immunospot Cat# CTLW-010 CellTrace Violet ThermoFisher Cat# C34571 Critical commercial assays COVID-SeroIndex, Kantaro Quantitative SARS-CoV-2 IgG Antibody Kit (Spike) R&D Systems Cat# DSR200 ARCHITECT SARS-CoV-2 IgG kit (Nucleocapsid) Abbot Cat# 6R86 ARCHITECT SARS-CoV-2 IgG II Quant (RBD) Abbot Cat# 6S60 Human IFN-g ELISpot plus kit Mabtech Cat# 3420-4APT IFNg-IL5 Fluorospot Mabtech Cat# FSP-0108 SARS-CoV-2 Spike B cell analysis kit Miltenyi Biotec Cat# 130-128-022 SARS-CoV-2 live neutralization assay Viroclinics Biosciences-Cerba N/A Pseudo-neutralization assay (D614 and D strains) Nexelis-Q2 solutions https://nexelis.com/our-expertise/ infectious-diseases/covid-19/ Pseudo-neutralization assay (D614) Monogram Biosciences-Labcorp https://monogrambio.labcorp.com/ phenosense-sars-cov-2neutralizing-antibody Software and algorithms Graphpad PRISM 7 GraphPad RRID:SCR_002798 FlowJo 10 BD Biosciences RRID:SCR_008520 SAS System software 9.4 SAS Institute Inc. RRID:SCR_008567 CTL Immunospot suite 2.7 Cellular Technology Limited RRID:SCR_011082 SPICE 6 NIH/NIAID https://niaid.github.io/spice/ RRID:SCR_016603 Article ll OPEN ACCESS

Techniques: Isolation, Enzyme-linked Immunospot, Negative Control, Variant Assay, Cytometry, Expressing, Functional Assay

Journal: Cell reports. Medicine

Article Title: GRAd-COV2 vaccine provides potent and durable humoral and cellular immunity to SARS-CoV-2 in randomized placebo-controlled phase 2 trial.

doi: 10.1016/j.xcrm.2023.101084

Figure Lengend Snippet: Figure 5. Long-term T and B memory response in the PBMC substudy (A) Kinetics of T cell response to SARS-CoV-2 spike at days 22 (post-dose 1-PD1), 36 (post-dose 2-PD2), and 180, evaluated by IFNg ELISpot and expressed as IFNg spot-forming cells (SFC) per million PBMCs. (B) Proliferative CD4 and CD8 T cell responses (percentage of CD4 or CD8 CellTrace low) following 5 day incubation with spike peptide pools, assessed in PBMCs collected at day 180 (day 36 for the four PL arm subjects). (C) Quantification of spike-specific (Spike++ as defined in STAR Methods) memory B cell (MBC) percentages in all 54 PBMC substudy subjects at day 36 and 180 visits. Data were analysed with two-tailed, paired Wilcoxon test, and only signifi- cant differences are shown. In all panels, gray symbols/lines indicate PL arm, while red and blue symbols/lines indicate SD and RD GRAd-COV2 arms, respectively. Open red symbols and pink shaded areas indicate volun- teers in the SD cohort that received an approved COVID-19 vaccine (defined SD+vax) between the day 57 and 180 visits. Horizontal black lines are set at GM.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Brilliant Stain Buffer BD Biosciences Cat# 563794 BD Fixation/Permeabilization Kit BD Biosciences Cat# 554714 FVS780 BD Biosciences Cat# 565388 HyClone Defined FBS, US Origin, 500 mL Cytiva Cat# SH30070.03 CTL-WashTM Supplement 10x Immunospot Cat# CTLW-010 CellTrace Violet ThermoFisher Cat# C34571 Critical commercial assays COVID-SeroIndex, Kantaro Quantitative SARS-CoV-2 IgG Antibody Kit (Spike) R&D Systems Cat# DSR200 ARCHITECT SARS-CoV-2 IgG kit (Nucleocapsid) Abbot Cat# 6R86 ARCHITECT SARS-CoV-2 IgG II Quant (RBD) Abbot Cat# 6S60 Human IFN-g ELISpot plus kit Mabtech Cat# 3420-4APT IFNg-IL5 Fluorospot Mabtech Cat# FSP-0108 SARS-CoV-2 Spike B cell analysis kit Miltenyi Biotec Cat# 130-128-022 SARS-CoV-2 live neutralization assay Viroclinics Biosciences-Cerba N/A Pseudo-neutralization assay (D614 and D strains) Nexelis-Q2 solutions https://nexelis.com/our-expertise/ infectious-diseases/covid-19/ Pseudo-neutralization assay (D614) Monogram Biosciences-Labcorp https://monogrambio.labcorp.com/ phenosense-sars-cov-2neutralizing-antibody Software and algorithms Graphpad PRISM 7 GraphPad RRID:SCR_002798 FlowJo 10 BD Biosciences RRID:SCR_008520 SAS System software 9.4 SAS Institute Inc. RRID:SCR_008567 CTL Immunospot suite 2.7 Cellular Technology Limited RRID:SCR_011082 SPICE 6 NIH/NIAID https://niaid.github.io/spice/ RRID:SCR_016603 Article ll OPEN ACCESS

Techniques: Enzyme-linked Immunospot, Incubation, Two Tailed Test