Journal: Frontiers in Immunology

Article Title: Presence of neutrophil extracellular traps (NETs) in different types of human urinary tract infections (UTI). A pilot study

doi: 10.3389/fimmu.2026.1745166

Figure Lengend Snippet: Detection of cathelicidin (LL-37) and myeloperoxidase (MPO) in human urine samples. Protein concentrations in human urine samples were detected by ELISA. The samples were analyzed in a microplate reader. Bar represents mean ± SD of each group. Protein levels were compared across groups (A, C) and subgroups (B, D). (A, C) LL-37 and MPO concentrations are higher in infected (Group A and B) than in healthy urines (Control). (B, D) No difference in protein concentrations was found when comparing the subgroups of Group A. ****p < 0.0001.

Article Snippet: For the MPO-specific ELISA kit, a 96-well high-binding polystyrene microplate (F-bottom, Greiner Bio-One, #655061) was used.

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Control

Journal: Nature Communications

Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection

doi: 10.1038/s41467-022-28898-1

Figure Lengend Snippet: Humoral immune responses were assessed in acute and convalescent by binding antibody ELISA for total IgG specific to the a Spike glycoprotein and b Nucleocapsid, quantification of c IgG memory B cells specific to the spike glycoprotein, and d pseudoneutralisation antibody titres. A two-tailed Wilcoxon rank-sum test was used to compare between study time points. The boxplots all display the median values with the first and third quartile, and the whiskers represent the highest and lowest values no more than 1.5 times the interquartile range from the corresponding hinge. A generalised additive mixed model (GAMM) by restricted maximum likelihood—right-hand plots—was used to fit the immunological measures (log10 transformed) taken at multiple study time points, using Gaussian process smooth term. The GAMM plots the ribbon represents the 95% confidence interval around the fitted value. Disease severity group was included in the GAMM as a linear predictor and a participant identifier was included as a random effect. See Table for number of individuals evaluated per assay.

Article Snippet: For detection of anti-spike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Transformation Assay

Journal: Nature Communications

Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection

doi: 10.1038/s41467-022-28898-1

Figure Lengend Snippet: SARS-CoV-2 spike-specific antibody isotype and subclasses measured post-infection: a IgM, b IgA, c IgG1 and d IgG3. Antibody function measure post-SARS-CoV-2 infection: e antibody-dependent NK cell activation (ADNKA), f antibody-dependent neutrophil phagocytosis (ADNP), g antibody-dependent monocyte phagocytosis (ADMP) and h antibody-dependent complement deposition (ADCD). i Polar plot of various antibody isotype, subclass and function data, minimum-maximum normalised. The boxplots all display the median values with the first and third quartile, and the whiskers represent the highest and lowest values no more than 1.5 times the interquartile range from the corresponding hinge. A two-tailed Wilcoxon rank-sum test was used to compare between study time points. A generalised additive mixed model (GAMM) by restricted maximum likelihood—right-hand plots—was used to fit the immunological measures (log10 transformed) taken at multiple study time points, using Gaussian process smooth term. The GAMM plots the ribbon represents the 95% confidence interval around the fitted value. Disease severity group was included in the GAMM as a linear predictor and a participant identifier was included as a random effect. See Table for number of individuals evaluated per assay.

Article Snippet: For detection of anti-spike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405.

Techniques: Infection, Activation Assay, Two Tailed Test, Transformation Assay

Journal: Immunity

Article Title: An interleukin-9-ZBTB18 axis promotes germinal center development of memory B cells.

doi: 10.1016/j.immuni.2025.02.021

Figure Lengend Snippet: Figure 3. ZBTB18 does not impinge on B cell activation or GC formation (A) Intracellular calcium responses of naive B cells from Cd79acre/+ and Cd79acre/+Zbtb18fl/flmice upon anti-IgM stimulation. Cells were pooled from 2 to 3 mice. Data represent three independent experiments. (B and C) Proliferation of Cd79acre/+ and Cd79acre/+Zbtb18fl/flnaive B cells 72 h after anti-IgM, LPS, or anti-CD40 stimulation. Shown are representative CellTrace Violet dilution profiles (B) and summary data of the fraction of divided cells (C). Each symbol represents one independent experiment with cells pooled from 2 to 3 mice per genotype. (D–H) The primary response of Aicda+/CreERT2;Zbtb18fl/fl;Rosa26-Ai14 and Aicda+/CreERT2;Zbtb18+/+;Rosa26-Ai14 mice. (D) The experimental design. Tamoxifen was given every other day starting from day 3. (E) Representative FACS profiles for gating GCs, GCMP cells, and NP-specific tdTomato+ MBCs. Fractional abundance of GCs in total B220+ cells (F), GCMP cells in GCs (G), and total MBCs or tdTomato+ MBCs in total B220+ cells (H). Data are pooled from 3 independent experiments.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Mouse: S1pr2-creERT2 Dr. T. Kurosaki; Osaka University N/A Mouse: B1-8hi Dr. M. Nussenzweig, Rockefeller University N/A Mouse: Cd79acre/+ Dr. M. Reth, University of Freiburg N/A Mouse: Il9-/- This paper N/A Mouse: Zbtb18fl/fl This paper N/A Mouse: Zbtb18Flag-in/Flag-in This paper N/A Oligonucleotides qPCR primers, see Table S1 This paper N/A ChIP-qPCR primers, see Table S1 This paper N/A Software and algorithms Adobe Illustrator Adobe https://www.adobe.com/products/illustrator.html Prism v8 Graphpad http://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 Tree Star https://www.flowjo.com/ Imaris Bitplane AG http://www.bitplane.com/imaris/imaris RRID:SCR_007370 Image J Wayne Rasband https://imagej.net/software/fiji/downloads Other BD FACS Aria III Cell Sorter BD Biosciences N/A LSR II cytometer BD Biosciences N/A iMark plate reader Bio-Rad Cat# 1681135 Aurora cytometer Cytek Biosciences N/A 96-well ELISA plates Costar Cat# 42592 CellDrop BF Brightfield Cell Counter DeNovix N/A Leica STELLARIS 8 Falcon confocal microscope Leica N/A Qubit 4 Fluorometer Invitrogen N/A Immunity 58, 1–14.e1–e6, April 8, 2025 e3 were immunized intraperitoneally with 10 mg NP-OVA (Biosearch Technologies) in PBS.

Techniques: Activation Assay

Journal: Immunity

Article Title: An interleukin-9-ZBTB18 axis promotes germinal center development of memory B cells.

doi: 10.1016/j.immuni.2025.02.021

Figure Lengend Snippet: Figure 5. ZBTB18 promotes survival of GCMP cells (A) The relative mRNA expression of Casp3 and Bid by qRT-PCR, with wild type set as 1. Each dot represents an independent sort of 200 cells. (B) Diagrams of 2-kb promoters of indicated gene, with ZBTB18-binding motifs highlighted and primers for ChIP-qPCR indicated by arrows. (C) Normalized luciferase activities, transcriptionally driven by 2-kb promoters with or without exogenous ZBTB18 transduced in 293T cells. Data are from 3 independent experiments, p values by t tests. (D) ChIP-qPCR analysis of ZBTB18-regulated apoptosis-promoting genes in MBCs from Zbtb18FLAG-in/FLAG-in mice. Shown are enrichment of promoter frag- ments, with immunoprecipitation performed using the anti-FLAG or control IgG antibody. One of two independent experiments with similar results is shown. (E) Representative FACS profiles (left) and summary data (right) of GCMP cells expressing active caspases. In scatter plots, each dot represents one mouse. Data are pooled from four independent experiments. p values by t tests, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, not significant.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Mouse: S1pr2-creERT2 Dr. T. Kurosaki; Osaka University N/A Mouse: B1-8hi Dr. M. Nussenzweig, Rockefeller University N/A Mouse: Cd79acre/+ Dr. M. Reth, University of Freiburg N/A Mouse: Il9-/- This paper N/A Mouse: Zbtb18fl/fl This paper N/A Mouse: Zbtb18Flag-in/Flag-in This paper N/A Oligonucleotides qPCR primers, see Table S1 This paper N/A ChIP-qPCR primers, see Table S1 This paper N/A Software and algorithms Adobe Illustrator Adobe https://www.adobe.com/products/illustrator.html Prism v8 Graphpad http://www.graphpad.com/ RRID:SCR_002798 FlowJo v10 Tree Star https://www.flowjo.com/ Imaris Bitplane AG http://www.bitplane.com/imaris/imaris RRID:SCR_007370 Image J Wayne Rasband https://imagej.net/software/fiji/downloads Other BD FACS Aria III Cell Sorter BD Biosciences N/A LSR II cytometer BD Biosciences N/A iMark plate reader Bio-Rad Cat# 1681135 Aurora cytometer Cytek Biosciences N/A 96-well ELISA plates Costar Cat# 42592 CellDrop BF Brightfield Cell Counter DeNovix N/A Leica STELLARIS 8 Falcon confocal microscope Leica N/A Qubit 4 Fluorometer Invitrogen N/A Immunity 58, 1–14.e1–e6, April 8, 2025 e3 were immunized intraperitoneally with 10 mg NP-OVA (Biosearch Technologies) in PBS.

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, ChIP-qPCR, Luciferase, Immunoprecipitation, Control