DOI: 10.6076/d10019
title: Antimicrobials from a feline skin commensal bacterium inhibit skin colonization and infection by drug-resistant S. pseudintermedius

description: Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is an
 important emerging zoonotic pathogen that causes severe skin infections.
 To combat infections from drug-resistant bacteria, the transplantation of
 commensal antimicrobial bacteria as a therapeutic has shown clinical
 promise. We screened a collection of diverse staphylococcus species from
 domestic dogs and cats for antimicrobial activity against MRSP. A unique
 strain (S. felis C4) was isolated from feline skin that inhibited MRSP and
 multiple gram-positive pathogens. Competition experiments in mice showed
 that S. felis significantly reduced MRSP skin colonization and an
 antimicrobial extract from S. felis significantly reduced necrotic skin
 injury from MRSP infection. Fluorescence and electron microscopy revealed
 that S. felis antimicrobials disrupted bacterial but not eukaryotic cell
 membranes. LC/MS identified several S. felis phenol-soluble modulin beta
 (PSMβ) peptides that exhibited antimicrobial and anti-inflammatory
 activity. These findings indicate a feline commensal bacterium that could
 be utilized in bacteriotherapy against difficult-to-treat animal and human
 skin infections.
description: NHEK cells were treated with DMSO (0.1%) control, PSMβ2 (10
 μg/ml), Poly I:C (0.4 μg/ml) or PSMβ2 and Poly I:C combined, all in
 triplicate wells, for 4 h or 24 h in and RNA was extracted using the
 PureLink RNA mini kit and triplicate samples were pooled together for each
 treatment. Isolated RNA was submitted to the UCSD IGM Genomics Center for
 RNA-sequencing performed on a high-output run V4 platform (Illumina, USA)
 with a single read 100 cycle runs. All the steps including trim bases,
 alignment with bowtie 2, quantification to annotation model (Partek E.M),
 Normalization (CPM/Add 1.0 E-4), Gene Set Enrichment (GSA), Differential
 Gene Analysis and hierarchial clustering, and Gene Ontology analysis was
 performed with the Partek Flow Genomics Suite.
description: contributor: Alan O'Neill                          
  <br> contributor: Nikhil Kulkarni                          
  <br>                                 <br> SAMPLES<br>
 Sample name    source name                          organism             
                                       characteristics: treatment         
                          characteristics: time            <br>
 Sample 1          Control_4_h_S76                  NHEK    Human primary
 keratinocytes    DMSO (0.1%)                                             
      4 h post-treatment            <br> Sample 2         
 Control_24_h_S80                NHEK    Human primary keratinocytes  
  DMSO (0.1%)                                                   24 h
 post-treatment            <br> Sample 3          Poly_IC_4_h_S78   
              NHEK    Human primary keratinocytes    Poly I:C (0.4 ug/ml) 
                                           4 h post-treatment          
  <br> Sample 4          Poly_IC_24_h_S82               NHEK    Human
 primary keratinocytes    Poly I:C (0.4 ug/ml)                             
             24 h post-treatment            <br> Sample 5         
 Poly_IC_PSMB_4_h_S79    NHEK    Human primary keratinocytes    Poly I:C
 (0.4 ug/ml) and PSMB2 (10 ug/ml)    4 h post-treatment          
  <br> Sample 6          Poly_IC_PSMB_24_h_S83  NHEK    Human primary
 keratinocytes    Poly I:C (0.4 ug/ml) and PSMB2 (10 ug/ml)    24 h
 post-treatment            <br> Sample 7          PSMB_4_h_S77       
             NHEK    Human primary keratinocytes     PSM?2 (10 ug/ml)     
                                        4 h post-treatment          
  <br> Sample 8          PSMB_24_h_S81                 NHEK    Human
 primary keratinocytes     PSM?2 (10 ug/ml)                               
              24 h post-treatment            <br>                   
              <br> PROTOCOLS<br> growth protocol: NHEK cells
 were cultured in EpiLife medium supplemented with EpiLife Defined Growth
 Supplement (EDGS)<br> treatment protocol: "NHEK cells were
 treated with DMSO (0.1%), PolyI:C (0.4 ?g/ml), PSM?2 (10 ?g/ml) or PolyI:C
 and PSM?2 for 4 h or 24 h."                            <br>
 extract protocol: NHEKs cells were washed 2X with PBS and RNA was
 extracted using PureLink RNA Mini Kit<br>                           
      <br> DATA PROCESSING PIPELINE<br> data processing step:
 "All the steps including trim bases, alignment with bowtie 2,
 quantification to annotation model (Partek E.M), Normalization (CPM/Add
 1.0 E-4), Gene Set Enrichment (GSA), Differential Gene Analysis and
 hierarchial clustering, and Gene Ontology analysis was performed with the
 Partek Flow Genomics Suite."                       <br> genome
 build: hg19                            <br> processed data files
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