Visualising virulence factors: Trichophyton benhamiaes subtilisins demonstrated in a guinea pig skin ex vivo model

Mycoses. 2020 Sep;63(9):970-978. doi: 10.1111/myc.13136. Epub 2020 Jul 15.

Abstract

Background: Dermatophytoses rank among the most frequent communicable diseases in humans, and the zoonotic transmission is increasing. The zoophilic dermatophyte Trichophyton (T.) benhamiae is nowadays one of the main causes of tinea faciei et corporis in children. However, scientific data on molecular pathomechanisms and specific virulence factors enabling this ubiquitous occurrence are scarce.

Objectives: To study tissue invasion and the expression of important virulence factors of T. benhamiae, isolates that were recovered from two groups of hosts (humans vs. guinea pigs (GP)) using an ex vivo skin model.

Methods: After confirmation of species identity by ITS sequencing, CFU suspensions of dermatophyte isolates (n = 20) were applied to the skin infection model and cultured. Employing specific immunofluorescence staining techniques, the expression of subtilisin 3 and 6 and metallocarboxypeptidase A was analysed. The general mode of invasion was explored. Results were compared with biopsies of naturally infected GP.

Results: All isolates were successfully recovered and proliferated well after application to the infection model. Progressive invasion of hyphae through all skin structures and destruction of explants were observed with early events being comparable to natural infection. An increasing expression of the examined virulence factors towards the end of culture was noticed but no difference between the two groups of isolates.

Conclusions: For the first time, important in vivo markers of dermatophytosis were visualised immunohistochemically in an ex vivo skin infection model and in skin biopsies of GP naturally infected with T. benhamiae. More research on the underlying pathomechanisms of dermatophyte infection is urgently needed.

Keywords: Trichophyton benhamiae; ex vivo model; guinea pig skin explants; immunofluorescence; metallocarboxypeptidase A; subtilisins; virulence factors; zoonotic dermatophytes.

MeSH terms

  • Animals
  • Cell Culture Techniques / methods*
  • Disease Models, Animal
  • Guinea Pigs
  • Humans
  • Organ Culture Techniques / methods
  • Skin / microbiology*
  • Skin / pathology
  • Tinea / microbiology*
  • Trichophyton / classification
  • Trichophyton / growth & development*
  • Trichophyton / pathogenicity*
  • Virulence Factors*
  • Visual Perception

Substances

  • Virulence Factors