title: Effect of Culture Supernatant Derived from Trichophyton Rubrum Grown in the Nail Medium on the Innate Immunity-related Molecules of HaCaT.
abstract: BACKGROUND
  Trichophyton rubrum is superficial fungi characteristically confined to dead keratinized tissues. These observations suggest that the soluble components released by the fungus could influence the host immune response in a cell in contact-free manner. Therefore, this research aimed to analyze whether the culture supernatant derived from T. rubrum grown in the nail medium could elicit the immune response of keratinocyte effectively.
  
  
  METHODS
  The culture supernatants of two strains (T1a, T XHB ) were compared for the β-glucan concentrations and their capacity to impact the innate immunity of keratinocytes. The β-glucan concentrations in the supernatants were determined with the fungal G-test kit and protein concentrations with bicinchoninic acid protein quantitative method, then HaCaT was stimulated with different concentrations of culture supernatants by adopting morphological method to select a suitable dosage. Expressions of host defense genes were assessed by quantitative polymerase chain reaction after the HaCaT was stimulated with the culture supernatants. Data were analyzed with one-way analysis of variance, followed by the least significant difference test.
  
  
  RESULTS
  The T. rubrum strains (T1a and T XHB ) released β-glucan of 87.530 ± 37.581 pg/ml and 15.747 ± 6.453 pg/ml, respectively into the media. The messenger RNA (mRNA) expressions of toll-like receptor-2 (TLR2), TLR4, and CARD9 were moderately up-regulated in HaCaT within 6-h applications of both supernatants. HaCaT cells were more responsive to T1a than T XHB . The slight increase of dendritic cells-specific intercellular adhesion molecule 3-grabbing nonintegrin expression was faster and stronger, induced by T1a supernatant than T XHB . The moderate decreases of RNase 7, the slight up-regulations of Dectin-1 and interleukin-8 at the mRNA level were detected only in response to T1a rather than T XHB . After a long-time contact, all the elevated defense genes decreased after 24 h.
  
  
  CONCLUSIONS
  The culture supernatant of T. rubrum could directly and transiently activate the innate immune response of keratinocytes.
authors.forename: Xin-Zhu
authors.initials: XZ
authors.lastname: Huang
authors.forename: Pan-Pan
authors.initials: PP
authors.lastname: Liang
authors.forename: Han
authors.initials: H
authors.lastname: Ma
authors.forename: Jin-Ling
authors.initials: JL
authors.lastname: Yi
authors.forename: Song-Chao
authors.initials: SC
authors.lastname: Yin
authors.forename: Zhi-Rui
authors.initials: ZR
authors.lastname: Chen
authors.forename: Mei-Rong
authors.initials: MR
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Lai
authors.affiliation: Department of Dermatology, the Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510630, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
mesh_terms: D045744:Cell Line, Tumor; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects*; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism*; D014249:Trichophyton / Q000378:metabolism*; D047071:beta-Glucans / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D047071:beta-Glucans