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, TXHB) 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 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 supernatants. HaCaT cells were more responsive to T1a than TXHB. The slight increase of dendritic cells-specific intercellular adhesion molecule 3-grabbing nonintegrin expression was faster and stronger, induced by T1a supernatant 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 supernatant of T. rubrum could directly and transiently activate the innate immune response of keratinocytes.

Keratitis–ichthyosis–deafness syndrome accompanied by disseminated cutaneous fungal infection

“…Our previous studies have already confirmed that TLR2 and TLR4 were involved in the immune response to T. rubrum infection. 8,9 In the present study, our results showed a suppressed native immune response in this patient with lower expression of TLR2 and no significant change of TLR4 at the protein level regardless of the presence of fungal infection or not, which might result in the susceptibility to T. rubrum infection in KID syndrome. Although the expression of TLR2 presented a small increase at the transcription level, it might be because some potential inhibitory/regulation factors in the KID syndrome patient influence the translation process of TLR2 protein.…”

“…Toll‐like receptors (TLR) are important pattern recognition receptors in mammals, and TLR2 and TLR4 play a crucial role in the host defense fungus infection . Our previous studies have already shown that keratinocyte can recognize T. rubrum , and induce native immune response through TLR2 and TLR4 . In the present study, we describe a rare Asian case of KID syndrome accompanied by disseminated cutaneous T. rubrum infection with mutation p.G12R of GJB2 .…”

“…6,7 Our previous studies have already shown that keratinocyte can recognize T. rubrum, and induce native immune response through TLR2 and TLR4. 8,9 In the present study, we describe a rare Asian case of KID syndrome accompanied by disseminated cutaneous T. rubrum infection with mutation p.G12R of GJB2. Then, we further explore the expression status of TLR2 and TLR4 in this patient to provide some insight into the mechanisms of why KID syndrome is susceptible to cutaneous fungal infection, and to offer some promising new therapeutic targets.…”

“…In the present study, we focused on TLR2 and TLR4, which are the very important pattern recognition receptors in the innate immune system, and explored the interaction between the innate immunity and the infectious complications of KID syndrome. Our previous studies have already confirmed that TLR2 and TLR4 were involved in the immune response to T. rubrum infection . In the present study, our results showed a suppressed native immune response in this patient with lower expression of TLR2 and no significant change of TLR4 at the protein level regardless of the presence of fungal infection or not, which might result in the susceptibility to T. rubrum infection in KID syndrome.…”

Extracellular Vesicles From the Dermatophyte Trichophyton interdigitale Modulate Macrophage and Keratinocyte Functions

“…A recent study showed that neither the IL-17 response nor adaptive immunity was required for fungal control of T. rubrum ( 35 ). Moreover, T. rubrum was reported to induce IL-1β production by macrophages, which, in turn, played essential roles in the immune response against T. rubrum ( 36 ), and the culture supernatant of T. rubrum was further shown to activate the innate immune response of HaCaT cells ( 37 ). The findings of these studies agree with our results showing that keratinocytes stimulated with EVs from T. interdigitale showed increased production of NO and proinflammatory mediators.…”