human pmn  (Brinkmann Instruments)

Journal: PLOS Pathogens

Article Title: Adenovirus phagocytosis by neutrophils triggers a pro-inflammatory response

doi: 10.1371/journal.ppat.1013504

Figure Lengend Snippet: (A) Ad cytotoxicity assessed via Sytox Green assay. PMNs were incubated with HBSS, Triton-X-100, human serum (HS) or HS-opsonized Ads (MOI 10 4 vp/cell) in the presence of the cell-impermeant fluorescent probe Sytox Green probe, a cell-impermeant DNA fluorescent probe. Cell fluorescence was measured with a plate reader. The curves represent the percentage (± SEM, 5 independent experiments) of PMN death ( i.e., PMN fluorescence relative to Triton-X-100-lysed PMN fluorescence multiplied by 100) at different incubation times. Kruskal-Wallis test was performed followed by Dunn’s multiple comparison tests: *, p < 0.05 (comparison with HS). (B) Ad-induced NET (neutrophil extracellular trap) formation. PMNs were incubated with buffer (control), PMA 100 nM (positive control) or HS-opsonized A488-Ad5 or Ad3 (MOI 10 4 vp/cell) for 4h. Then, after cell fixation, cells were permeabilized, immunolabeled for myeloperoxidase (MPO) and stained with DAPI. The images are Z-projections generated from Z-stacks. Three independent experiments were performed. Scale bar = 10 µm.

Article Snippet: For the second method, PMNs were isolated using the MACSxpress Whole Blood Neutrophil Isolation Kit (Miltenyi Biotec, 130-104-434) and then the MACSxpress Erythrocyte Depletion Kit (Miltenyi Biotec, 130-098-196) making it possible to discard the remaining red blood cells.

Techniques: Incubation, Fluorescence, Comparison, Control, Positive Control, Immunolabeling, Staining, Generated

Journal: Cell reports

Article Title: Staphylococcus aureus promotes strain-dependent immunopathology during cutaneous leishmaniasis through induction of IL-1β.

doi: 10.1016/j.celrep.2025.115624

Figure Lengend Snippet: Figure 2. S. aureus triggers IL-1β to establish early inflammation and pathology during an L. major infection (A) Pathology scoring and ear thickness measurements in mice uninfected or infected with either L. major alone or with both S. aureus colonization and L. major infection (Wilcoxon, *p < 0.05 and **p < 0.01; mean ± SD representative of 2 experiments; L. major-S. aureus treatment, IL-1β−/−mouse, n = 8 mice, and all other conditions, n = 6 mice). (B) Bacterial burden (CFUs) and L. major parasite burden at day 7 post-L. major infection (Wilcoxon, *p < 0.05). (C) Flow cytometric analysis of immune cell recruitment (Ly6G+, CD4+, and CD8+ cells) in infected ear skin (Wilcoxon, *p < 0.05, **p < 0.01, and ***p < 0.001; mean representative of 2 experiments; L. major-S. aureus treatment, IL-1β−/−mouse, n = 8 mice and all other conditions, n = 6 mice). (D) Percentage of immune cells recruited to the site of infection in mice infected with L. major with and without colonization of S. aureus. (E) Pie chart showing percentage of IL-1β+ cells that co-stain with neutrophils, macrophages, monocytes, and unidentified cells at 7 days post-infection. Based on flow cytometric data.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Staphylococcus aureus CLSA51 this paper CLSA51 Staphylococcus aureus CLSA52 this paper CLSA52 Staphylococcus aureus CLSA54 this paper CLSA54 Staphylococcus aureus CLSA56 this paper CLSA56 Staphylococcus aureus CLSA57 this paper CLSA57 Staphylococcus aureus CLSA62 this paper CLSA62 Chemicals, peptides, and recombinant proteins blood agar Remel Cat#R01201 Tryptic soy broth, Difco Fisher Scientific Cat#DF0370 RPMI 1640 Invitrogen Cat#11875085 Liberase TL Roche Cat#540102000 Dnase I Sigma Aldrich Cat#DN25 Neutrophil isolation medium (Lympholytepoly cell separation media) Cedarlane Cat#CL5070 Red blood cell lysis buffer Roche Cat#11814389001 Trizol Ambion Inc Cat#15596026 BBL CHROMagar Staph aureus BD Cat#214982 Saponin 10% Sigma Cat#59700 Chocolate agar Remel Cat#R08240 Microbank microbial storage tubes Pro-lab Cat#PL.170C/M Schneiders Drosophila medium GIBCO Cat#21720024 Fetal Bovine Serum Avantor Cat#89510186 L-glutamine 200mM Sigma Cat#G7513 Ficoll 400 Sigma Cat#F2637 10% Rat IgG Sigma Cat#18015 Formaldehyde Aqueous Solution Electron Microscopy Sciences Cat#15700 Leukocyte Activation Cocktail BD Biosciences Cat#550583 Covaris milliTube 2mL Covaris Cat#520132 Covaris tissueTUBE TT05M Covaris Cat#520139 Percoll Cytiva Cat#17089101 ACK lysing buffer Quality Biological Cat#118156721 Penicillin-Streptomycin Sigma Cat#P0781 Critical commercial assays Quick DNA mini-prep plus kit Zymo research Cat#D4069 DNAase digestion Qiagen Cat#79294 2100 Bioanalyzer and Eukaryotic Total RNA 6000 Pico Kit Agilent Cat#5067-1513 Qubit by RNA Broad Range assay Invitrogen Cat#Q10210 Illumina Stranded Total RNA Prep with Ribo-Zero Plus Illumina Cat#20040525 ELISA MAXTM Deluxe Set Human IL-1β BioLegend Cat#437004 Deposited data Whole genome sequences of 30 S aureus clinical isolates this paper NCBI Bioproject: PRJNA922957 Murine RNAseq data this paper NCBI Bioproject: PRJNA1212200 Human RNAseq data Farias Amorim et al.9 NCBI Bioproject: PRJNA885131 Experimental models: Organisms/strains C57BL6/J Mice The Jackson Laboratory Cat#000664 C57BL6/J IL-1b− /− Mice Dr.

Techniques: Infection, Staining

Journal: Cell reports

Article Title: Staphylococcus aureus promotes strain-dependent immunopathology during cutaneous leishmaniasis through induction of IL-1β.

doi: 10.1016/j.celrep.2025.115624

Figure Lengend Snippet: Figure 3. Pro-inflammatory pathways associated with IL-1β, cytotoxicity, cell death, and neutrophil recruitment are enriched in human Leishmania infections complicated by S. aureus (A) Schematic outlining the study design of sample collection from patients with L. braziliensis infections. (B) Gene expression of IL-1β in lesion biopsies with an S. aureus-dominant or heterogeneous microbiome profile (Wilcoxon, **p < 0.01). (C) Relative neutrophils in lesion biopsies as determined by cell estimation (XCell) (Wilcoxon, *p < 0.05). (D) GO pathway analysis comparing lesion biopsies with S. aureus dominance or a heterogeneous microbiome (Fisher’s one-tailed test with Benjamini-Hochberg adjustment, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Staphylococcus aureus CLSA51 this paper CLSA51 Staphylococcus aureus CLSA52 this paper CLSA52 Staphylococcus aureus CLSA54 this paper CLSA54 Staphylococcus aureus CLSA56 this paper CLSA56 Staphylococcus aureus CLSA57 this paper CLSA57 Staphylococcus aureus CLSA62 this paper CLSA62 Chemicals, peptides, and recombinant proteins blood agar Remel Cat#R01201 Tryptic soy broth, Difco Fisher Scientific Cat#DF0370 RPMI 1640 Invitrogen Cat#11875085 Liberase TL Roche Cat#540102000 Dnase I Sigma Aldrich Cat#DN25 Neutrophil isolation medium (Lympholytepoly cell separation media) Cedarlane Cat#CL5070 Red blood cell lysis buffer Roche Cat#11814389001 Trizol Ambion Inc Cat#15596026 BBL CHROMagar Staph aureus BD Cat#214982 Saponin 10% Sigma Cat#59700 Chocolate agar Remel Cat#R08240 Microbank microbial storage tubes Pro-lab Cat#PL.170C/M Schneiders Drosophila medium GIBCO Cat#21720024 Fetal Bovine Serum Avantor Cat#89510186 L-glutamine 200mM Sigma Cat#G7513 Ficoll 400 Sigma Cat#F2637 10% Rat IgG Sigma Cat#18015 Formaldehyde Aqueous Solution Electron Microscopy Sciences Cat#15700 Leukocyte Activation Cocktail BD Biosciences Cat#550583 Covaris milliTube 2mL Covaris Cat#520132 Covaris tissueTUBE TT05M Covaris Cat#520139 Percoll Cytiva Cat#17089101 ACK lysing buffer Quality Biological Cat#118156721 Penicillin-Streptomycin Sigma Cat#P0781 Critical commercial assays Quick DNA mini-prep plus kit Zymo research Cat#D4069 DNAase digestion Qiagen Cat#79294 2100 Bioanalyzer and Eukaryotic Total RNA 6000 Pico Kit Agilent Cat#5067-1513 Qubit by RNA Broad Range assay Invitrogen Cat#Q10210 Illumina Stranded Total RNA Prep with Ribo-Zero Plus Illumina Cat#20040525 ELISA MAXTM Deluxe Set Human IL-1β BioLegend Cat#437004 Deposited data Whole genome sequences of 30 S aureus clinical isolates this paper NCBI Bioproject: PRJNA922957 Murine RNAseq data this paper NCBI Bioproject: PRJNA1212200 Human RNAseq data Farias Amorim et al.9 NCBI Bioproject: PRJNA885131 Experimental models: Organisms/strains C57BL6/J Mice The Jackson Laboratory Cat#000664 C57BL6/J IL-1b− /− Mice Dr.

Techniques: Gene Expression, One-tailed Test

Journal: Cell reports

Article Title: Staphylococcus aureus promotes strain-dependent immunopathology during cutaneous leishmaniasis through induction of IL-1β.

doi: 10.1016/j.celrep.2025.115624

Figure Lengend Snippet: Figure 4. S. aureus isolates cultured from human cutaneous leishmaniasis lesions are phylogenetically and phenotypically diverse (A) Schematic of bacterial swab extraction and isolate identification by MALDI-TOF. (B) Quantification of the frequency of bacterial species recovered from 44 human cutaneous leishmaniasis lesions. Each species is only counted once per lesion. (C) Phylogenetic tree of S. aureus clinical isolates and reference genomes. Layered onto the tree is each isolate’s methicillin resistance status and clonal complex. (D) ELISA quantifying levels of IL-1β recovered from mouse and human neutrophils infected with CLSA2 and CLSA50 strains (Wilcoxon, ***p < 0.001). (E) Bacterial survival of CLSA2 and CLSA50 in response to infection of human and mouse neutrophils for 1 h (paired t test, *p < 0.05). n = 4 human donors; n = 5 mice.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Staphylococcus aureus CLSA51 this paper CLSA51 Staphylococcus aureus CLSA52 this paper CLSA52 Staphylococcus aureus CLSA54 this paper CLSA54 Staphylococcus aureus CLSA56 this paper CLSA56 Staphylococcus aureus CLSA57 this paper CLSA57 Staphylococcus aureus CLSA62 this paper CLSA62 Chemicals, peptides, and recombinant proteins blood agar Remel Cat#R01201 Tryptic soy broth, Difco Fisher Scientific Cat#DF0370 RPMI 1640 Invitrogen Cat#11875085 Liberase TL Roche Cat#540102000 Dnase I Sigma Aldrich Cat#DN25 Neutrophil isolation medium (Lympholytepoly cell separation media) Cedarlane Cat#CL5070 Red blood cell lysis buffer Roche Cat#11814389001 Trizol Ambion Inc Cat#15596026 BBL CHROMagar Staph aureus BD Cat#214982 Saponin 10% Sigma Cat#59700 Chocolate agar Remel Cat#R08240 Microbank microbial storage tubes Pro-lab Cat#PL.170C/M Schneiders Drosophila medium GIBCO Cat#21720024 Fetal Bovine Serum Avantor Cat#89510186 L-glutamine 200mM Sigma Cat#G7513 Ficoll 400 Sigma Cat#F2637 10% Rat IgG Sigma Cat#18015 Formaldehyde Aqueous Solution Electron Microscopy Sciences Cat#15700 Leukocyte Activation Cocktail BD Biosciences Cat#550583 Covaris milliTube 2mL Covaris Cat#520132 Covaris tissueTUBE TT05M Covaris Cat#520139 Percoll Cytiva Cat#17089101 ACK lysing buffer Quality Biological Cat#118156721 Penicillin-Streptomycin Sigma Cat#P0781 Critical commercial assays Quick DNA mini-prep plus kit Zymo research Cat#D4069 DNAase digestion Qiagen Cat#79294 2100 Bioanalyzer and Eukaryotic Total RNA 6000 Pico Kit Agilent Cat#5067-1513 Qubit by RNA Broad Range assay Invitrogen Cat#Q10210 Illumina Stranded Total RNA Prep with Ribo-Zero Plus Illumina Cat#20040525 ELISA MAXTM Deluxe Set Human IL-1β BioLegend Cat#437004 Deposited data Whole genome sequences of 30 S aureus clinical isolates this paper NCBI Bioproject: PRJNA922957 Murine RNAseq data this paper NCBI Bioproject: PRJNA1212200 Human RNAseq data Farias Amorim et al.9 NCBI Bioproject: PRJNA885131 Experimental models: Organisms/strains C57BL6/J Mice The Jackson Laboratory Cat#000664 C57BL6/J IL-1b− /− Mice Dr.

Techniques: Cell Culture, Extraction, Enzyme-linked Immunosorbent Assay, Infection