Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: S. hominis makes six AIP variants. (A) Schematic of the conserved staphylococcal accessory gene regulator quorum sensing system. (B) Predicted agrD sequences for each S. hominis agr type identified by PCR. Nonconserved residues are marked in red. The Clustal Omega sequence alignment between all strains is shown at the bottom, with asterisks representing fully conserved residues, colons representing residues with strongly similar properties, and periods representing residues with weakly similar properties. The confirmed peptide sequence of each AIP is underlined. (C) Representative images of predicted AIP structures for each agr type. Previously confirmed structures are marked with an asterisk.

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Sequencing

Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: S. hominis conditioned medium (CM) inhibits MRSA agr signaling. MRSA agr -I P3::YFP (A), agr -II P3::YFP (B), agr -III P3::YFP (C), and agr -IV P3::YFP (D) reporter strains were incubated with 10% cell-free CM from S. hominis strains for 24 h. The 24-h fluorescent point (relative fluorescence units [RFU]) is shown relative to the reporter-only control. Data were pooled from three independent experiments. Significance was determined by ordinary one-way ANOVA with Dunnett’s multiple-comparison test (degree of freedom [df] = 32, F = 12.00 [A], 12.56 [B], 21.65 [C], 1.78 [D]). Mean values ± SD are shown. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. NS, not significant.

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Incubation, Fluorescence, Control, Comparison

Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: Identification and validation of three novel S. hominis AIPs. The amino acid sequence and thiolactone structure of S. hominis AIP-II (A), AIP-IV (B), and AIP-V (C) were solved and confirmed using MS-MS analysis. Characteristic y ions are listed for each AIP. The calculated mass of the protonated peptide (Calc.) is shown beneath each structure in comparison to the experimental mass (Exp.) and mass error.

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Biomarker Discovery, Sequencing, Tandem Mass Spectroscopy, Comparison

Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: Synthetic S. hominis AIPs inhibit MRSA agr signaling. (A to E) MRSA P3::YFP reporters were incubated with increasing doses of S. hominis synthetic AIP-I (A), AIP-II (B), AIP-III (C), AIP-IV (D), or AIP-V (E). Fluorescent intensity relative to the reporter only at the 24-h time point is shown. (F) IC 50 values (nM) were calculated for each AIP-reporter pair from a four-parameter nonlinear regression curve. The 95% confidence interval for each IC 50 value is reported in parentheses. Results were pooled from three independent experiments. Mean ± SD values are shown. NC, not calculated.

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Incubation

Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: S. hominis synthetic AIPs inhibit S. epidermidis agr signaling. (A to E) S. epidermidis P3::sGFP reporters were incubated with increasing doses of S. hominis synthetic AIP-I (A), AIP-II (B), AIP-III (C), AIP-IV (D), or AIP-V (E). Fluorescent intensity relative to the reporter only at the 24-h time point is shown. (F) IC 50 values (nM) were calculated for each AIP-reporter pair from a four-parameter nonlinear regression curve. The 95% confidence interval for each IC 50 value is reported in parentheses. Results are pooled from three independent experiments. Mean ± SD values are shown. NC, not calculated.

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Incubation

Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: S. hominis agr- I and agr -II are the most abundant agr types on healthy human skin. (A) Relative frequency of S. hominis agr types identified in all published S. hominis genomes in the NCBI database. (B) Absolute abundance by quantitative PCR of S. hominis agr types in a 5-cm 2 swab of the antecubital crease pooled from 14 healthy skin swabs. (C) Relative abundance of each S. hominis agr type stratified by patient swab.

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Real-time Polymerase Chain Reaction

Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: S. hominis agr -I and agr -II intraspecies cross talk. (A) Relative fluorescent values of S. hominis agr -I P3::YFP incubated with increasing doses of synthetic AIP-I at 8 h of incubation. (B) Relative fluorescent values of S. hominis agr -I P3::YFP incubated with increasing doses of synthetic AIP-II at 8 h of incubation. (C) Relative fluorescent values of S. hominis agr -II P3::YFP incubated with increasing doses of synthetic AIP-II at 8 h of incubation. (D) Relative fluorescent values of S. hominis agr -II P3::YFP incubated with increasing doses of synthetic AIP-I at 8 h of incubation. Results were pooled from three independent experiments. Significance was determined by ordinary one-way ANOVA with Dunnett’s multiple-comparison test (df = 20, F = 8.53 [A], 9.75 [B], 6.40 [C], 17.23 [D]). Mean values ± SD are shown. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Incubation, Comparison

Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: S. hominis AIP-II protects murine skin from MRSA damage. (A) Dermonecrotic lesion size for mice administered a 10- or 50-μg dose of synthetic S. hominis AIP-II compared to that of mice administered wild type (WT) plus vehicle (DMSO) or Δ agr mutant plus vehicle control. (B) Weight change for indicated groups. (C) Representative images of dermonecrotic lesion size 5 days postinfection. Data were pooled from two independent experiments ( n = 10 per group). Mean ± SEM is shown. Significance was determined by two-way ANOVA with Dunnett’s multiple-comparison test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (D) Transepithelial water loss 72 h postinfection for mice administered a 10- or 50-μg dose of synthetic S. hominis AIP-II compared to that of mice administered WT plus vehicle (DMSO) or Δ agr mutant plus vehicle control. (E) Total CFU burden in epicutaneous lesions at 72 h postinfection (hpi) for the indicated groups. (F) Representative images of epicutaneous damage at 72 hpi. Data were pooled from two independent experiments ( n = 10 per group). Mean ± SD is shown. Significance was determined by ordinary one-way ANOVA with Dunnett’s multiple-comparison test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. N.S., not significant.

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Mutagenesis, Control, Comparison

Journal: mBio

Article Title: The Ubiquitous Human Skin Commensal Staphylococcus hominis Protects against Opportunistic Pathogens

doi: 10.1128/mbio.00930-22

Figure Lengend Snippet: Genes under transcriptional control of S. hominis AgrA. (A) RNA-seq heat map of genes with greater than a 4-fold change in expression and a false discovery rate P value of less than 0.05 in 100 μM apicidin-treated S. hominis compared to wild-type vehicle (DMSO)-treated cells. Fold changes are indicated in each box. Numbers in italics refer to the last five digits of the locus tag. (B) Mass spectrometric validation of the absence of PSM-β1 production in S. hominis treated with 100 μM apicidin. Results are averages from triplicate injections of biological triplicates (9 independent injections). The PSM-β1 predicted sequence and locus tag number are shown above the graph. The ion detected was at m/z 908.6785, which corresponds to the predicted mass (908.6819 Δm = 3.7 ppm) of PSM-β1with five protons and a +5 charge ([M + 5H] 5+ ).

Article Snippet: S. hominis synthetic AIPs were custom synthesized by AnaSpec, Inc.

Techniques: Control, RNA Sequencing, Expressing, Biomarker Discovery, Sequencing