human pmn  (Brinkmann Instruments)

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

Journal: Frontiers in Molecular Biosciences

Article Title: Development of an enzyme-linked immunosorbent assay (ELISA) for determining neutrophil elastase (NE) – a potential useful marker of multi-organ damage observed in COVID-19 and post-Covid-19 (PCS)

doi: 10.3389/fmolb.2025.1542898

Figure Lengend Snippet: Scheme of the designed ELISA assay for the quantitative assessment of neutrophil elastase in blood serum.

Article Snippet: NE concentration was determined using an ELISA kit (Human PMN (Neutrophil) Elastase ELISA Kit, ThermoFisher Scientific, Massachusetts, GA, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Development of an enzyme-linked immunosorbent assay (ELISA) for determining neutrophil elastase (NE) – a potential useful marker of multi-organ damage observed in COVID-19 and post-Covid-19 (PCS)

doi: 10.3389/fmolb.2025.1542898

Figure Lengend Snippet: Absorbance values for the individual concentrations of the standard (human native neutrophil elastase) – samples 1-22, and for the background (BLANK); standard number: 1-22 - in the order of increasing standard dilutions. The main steps of the ELISA test were carried out under the following conditions: I) Incubation with AbI: 440x, 2.5 h, 37°C; II) blocking solutions: 5% skimmed milk, overnight, 4°C; III) AbII: 5000x, 1.5 h, 37°C.

Article Snippet: NE concentration was determined using an ELISA kit (Human PMN (Neutrophil) Elastase ELISA Kit, ThermoFisher Scientific, Massachusetts, GA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Blocking Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Development of an enzyme-linked immunosorbent assay (ELISA) for determining neutrophil elastase (NE) – a potential useful marker of multi-organ damage observed in COVID-19 and post-Covid-19 (PCS)

doi: 10.3389/fmolb.2025.1542898

Figure Lengend Snippet: Standard curve for quantitative assessment of neutrophil elastase in blood serum (for concentration range of standard: 20 pg/mL to 300 pg/mL) under the given ELISA test conditions of the main steps: I) Incubation with AbI: 440x, 2.5 h, 37°C; II) blocking solutions: 5% skimmed milk, overnight, 4°C; III) AbII: 5000x, 1.5 h, 37°C.

Article Snippet: NE concentration was determined using an ELISA kit (Human PMN (Neutrophil) Elastase ELISA Kit, ThermoFisher Scientific, Massachusetts, GA, USA).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Incubation, Blocking Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Development of an enzyme-linked immunosorbent assay (ELISA) for determining neutrophil elastase (NE) – a potential useful marker of multi-organ damage observed in COVID-19 and post-Covid-19 (PCS)

doi: 10.3389/fmolb.2025.1542898

Figure Lengend Snippet: Standard curve for quantitative assessment of neutrophil elastase, corrected with control values, in the concentration range of standard: 1.5 pg/μL to 50 pg/μL. The main steps of the ELISA test were carried out under the following conditions: I) Incubation with AbI: 440x, 2.5 h, 37°C; II) blocking solutions: 7.5% skimmed milk, overnight, 4°C; III) AbII: 5000x, 1.5 h, 37°C.

Article Snippet: NE concentration was determined using an ELISA kit (Human PMN (Neutrophil) Elastase ELISA Kit, ThermoFisher Scientific, Massachusetts, GA, USA).

Techniques: Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Incubation, Blocking Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Development of an enzyme-linked immunosorbent assay (ELISA) for determining neutrophil elastase (NE) – a potential useful marker of multi-organ damage observed in COVID-19 and post-Covid-19 (PCS)

doi: 10.3389/fmolb.2025.1542898

Figure Lengend Snippet: Standard curve for human native neutrophil elastase determined based on absorbance values obtained using primary antibodies diluted 440 times, in the concentration range of standard: 14 pg/μL to 200 pg/μL. The main steps of the ELISA test were carried out under the following conditions: I) Incubation with AbI: 440x, 2.5 h, 37°C; II) blocking solutions: 7.5% skimmed milk, overnight, 4°C; III) AbII: 5000x, 1.5 h, 37°C.

Article Snippet: NE concentration was determined using an ELISA kit (Human PMN (Neutrophil) Elastase ELISA Kit, ThermoFisher Scientific, Massachusetts, GA, USA).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Incubation, Blocking Assay