RESUMEN
Objective To explore the expressions of Toll-like receptors (TLRs) 2 and 4 in mouse skin during early immune responses against Sporothrix.Methods A total of 60 BALB/c mice were randomly and equally divided into an experimental group and a control group to be intracutaneously injected with Sporothrix conidium suspensions at a concentration of 1 × 106 cfu/ml and sodium chloride physiological solution respectively.Five mice were sacrificed before the injection,and at 6,12,24,48,and 96 hours after the injection in each group,blood samples were obtained from the mice before sacrifice,and skin tissue specimens were resected from the area around the injection sites after sacrifice.Realtime fluorescence-based quantitative PCR was performed to quantify the mRNA expressions of TLR2 and TLR4,and immunohistochemical staining to observe the protein expressions of TLR2 and TLR4 in mouse skin specimens.Enzymelinked immunosorbent assay (ELISA) was conducted to determine the levels of interleukin 12 (IL-12) and tumor necrosis factor α (TNFα) in serum samples from the mice.Results After injection of Sporothrix conidium suspensions,the mRNA expression level of TLR2 gradually increased and peaked at 24 hours,which was 18.8 times that in the control group at 6 hours and 34 times at 24 hours.In addition,the mRNA expression level of TLR4 in the experiment group reached a peak,and was 56.7 times that in the control group at 6 hours after injection,then gradually decreased and reached the nadir at 96 hours.As immunohistochemical staining revealed,TLR2 and TLR4 were apparently expressed in both keratinocytes and macrophages in skin specimens from the experimental group,but not obviously in those from the control group.No significant differences were observed between the experimental group and control group in serum levels of IL-12 or TNF-α at any of the sampling time points.Conclusion TLR2 and TLR4 may play a favoring role in immunological defense by participating in the recognition of Sporothrix by keratinocytes and macrophages in mouse skin.
RESUMEN
We evaluated the differential expression and function of chitinase 3‐like‐1 in macrophage stimulated by Sporothrix schenckii and Candida albicans fungicidal ability of macrophage after stimulation with Sporothrix schenckii and Candida albi‐cans separately was detected .The expression of CHI3L1 gene in macrophage stimulated by Sporothrix Schenckii and Candida albicans was evaluated with real‐time PCR .The function of CHI3L1 protein in macrophages against the reproduction of Sporo‐thrix schenckii and Candida albicans was detected in vitro .Results showed that macrophages could engulf and kill Sporothrix Schenckii and Candida albicans in vitro .The expression of CHI3L1 gene in macrophage stimulated by Candida albicans was increased obviously .At the same time ,CHI3L1 protein can damper the reproduction of Candida albicans .However ,the ex‐pression of CHI3L1 gene was not elevated when macrophage was stimulated by Sporothrix schenckii and CHI3L1 protein played little role in reproduction of Sporothrix schenckii .The expression of CHI3L1 gene in macrophage was elevated after stimulation with Candida albicans ,but was not elevated with Sporothrix Schenckii .In correspondence with differential ex‐pression ,CHI3L1 in macrophages could impair the reproduction of Candida albicans but had a weak function on Sporothrix schenckii which might contribute to the pathogenesis of spo‐rotricosis .