title: Nanovesicles from Malassezia sympodialis and host exosomes induce cytokine responses--novel mechanisms for host-microbe interactions in atopic eczema.
abstract: BACKGROUND
  Intercellular communication can occur via the release of membrane vesicles. Exosomes are nanovesicles released from the endosomal compartment of cells. Depending on their cell of origin and their cargo they can exert different immunoregulatory functions. Recently, fungi were found to produce extracellular vesicles that can influence host-microbe interactions. The yeast Malassezia sympodialis which belongs to our normal cutaneous microbial flora elicits specific IgE- and T-cell reactivity in approximately 50% of adult patients with atopic eczema (AE). Whether exosomes or other vesicles contribute to the inflammation has not yet been investigated.
  
  
  OBJECTIVE
  To investigate if M. sympodialis can release nanovesicles and whether they or endogenous exosomes can activate PBMC from AE patients sensitized to M. sympodialis.
  
  
  METHODS
  Extracellular nanovesicles isolated from M. sympodialis, co-cultures of M. sympodialis and dendritic cells, and from plasma of patients with AE and healthy controls (HC) were characterised using flow cytometry, sucrose gradient centrifugation, Western blot and electron microscopy. Their ability to stimulate IL-4 and TNF-alpha responses in autologous CD14, CD34 depleted PBMC was determined using ELISPOT and ELISA, respectively.
  
  
  RESULTS
  We show for the first time that M. sympodialis releases extracellular vesicles carrying allergen. These vesicles can induce IL-4 and TNF-α responses with a significantly higher IL-4 production in patients compared to HC. Exosomes from dendritic cell and M. sympodialis co-cultures induced IL-4 and TNF-α responses in autologous CD14, CD34 depleted PBMC of AE patients and HC while plasma exosomes induced TNF-α but not IL-4 in undepleted PBMC.
  
  
  CONCLUSIONS
  Extracellular vesicles from M. sympodialis, dendritic cells and plasma can contribute to cytokine responses in CD14, CD34 depleted and undepleted PBMC of AE patients and HC. These novel observations have implications for understanding host-microbe interactions in the pathogenesis of AE.
authors.affiliation: Clinical Allergy Research Unit, Department of Medicine Solna, Karolinska Institutet, Stockholm, Sweden. ulf.gehrmann@ki.se
authors.forename: Ulf
authors.initials: U
authors.lastname: Gehrmann
authors.forename: Khaleda Rahman
authors.initials: KR
authors.lastname: Qazi
authors.forename: Catharina
authors.initials: C
authors.lastname: Johansson
authors.forename: Kjell
authors.initials: K
authors.lastname: Hultenby
authors.forename: Maria
authors.initials: M
authors.lastname: Karlsson
authors.forename: Lena
authors.initials: L
authors.lastname: Lundeberg
authors.forename: Susanne
authors.initials: S
authors.lastname: Gabrielsson
authors.forename: Annika
authors.initials: A
authors.lastname: Scheynius
mesh_terms: D000328:Adult; D000485:Allergens / Q000276:immunology; D016022:Case-Control Studies; D018920:Coculture Techniques; D016207:Cytokines / Q000096:biosynthesis*; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D003876:Dermatitis, Atopic / Q000276:immunology / Q000382:microbiology* / Q000473:pathology; D055354:Exosomes / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D054884:Host-Pathogen Interactions / Q000276:immunology*; D006801:Humans; D056747:Immunomodulation; D015847:Interleukin-4 / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D008290:Malassezia / Q000166:cytology* / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology; D049329:Nanostructures / Q000382:microbiology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D055815:Young Adult
chemical_list: D000485:Allergens; D016207:Cytokines; D014409:Tumor Necrosis Factor-alpha; D015847:Interleukin-4