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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 supematant derived from T.rubrum grown in the nail medium could elicit the immune response of keratinocyte effectively.Methods: The culture supematants of two strains (T1a, TXHB) were compared for the β-glucan concentrations and their capacity to impact the innate immunity of keratinocytes.The β-glucan concentrations in the supatants 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 supatants 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 supatants.Data were analyzed with one-way analysis of variance, followed by the least significant difference test.Results: The T.rubrum strains (T1 a and TXHB) 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 supatants.HaCaT cells were more responsive to T1 a than TXHB.The slight increase of dendritic cells-specific intercellular adhesion molecule 3-grabbing nonintegrin expression was faster and stronger, induced by T1 a supatant than TXHB.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 TXHB.After a long-time contact, all the elevated defense genes decreased after 24 h.Conclusion: The culture supatant of T.rubrum could directly and transiently activate the innate immune response of keratinocytes.