Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Adhesion to extracellular matrix components and HeLa cells of Sporothrix schenckii and Sporothrix brasiliensis PAP1 -silenced strains. The indicated extracellular matrix component was used to coat 96-well plates, then yeast-like cells were added, unbound cells were removed by extensive washing, and adherent cells were detected by ELISA with a primary anti-rHsp60 antibody. Control refers to wells coated with bovine serum albumin. Panel A contains data generated with S. schenckii, and WT is the 1099–18 ATCC MYA 4821 strain. Panel B shows results with S. brasiliensis cells, and the WT is the 5110 ATCC MYA 4823 strain. Alternatively, for both panels, 1 × 10 6 HeLa cells were placed per well, incubated 24 h at 37 °C and 5 % ( v /v) CO 2 , and used in the adhesion assays. Results are means ± SD of three biological replicates performed in duplicate. The Dunnett's test and then the unpaired t-test were used for data analysis. In both panels, * P < 0.05 when compared to WT or strains HSB1 and HSB2.

Article Snippet: HSB32 , 5110 ATCC MYA 4823 transformed with pCambia-Nou-PAP1 , 95.7 ± 3.2 , 1.1 ± 0.4 , This work.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Generated, Incubation

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Biofilm formation in Sporothrix schenckii and Sporothrix brasiliensis PAP1 -silenced strains. Biofilms were generated in microplates, using yeast-like cells. Non-adherent cells were removed, and biofilms were allowed to mature for 24 h at 37 °C. Biomass was stained with crystal violet and quantified by spectrophotometry. Panel A shows data with S. schenckii cells, and the WT is 1099–18 ATCC MYA 4821 strain. Panel B contains results obtained with S. brasiliensis cells, and the WT is the 5110 ATCC MYA 4823 strain. Data are means ± SD of three biological replicates. Results were analyzed with Dunnett's test and then the t-test. * P < 0.05 when compared to WT.

Article Snippet: HSB32 , 5110 ATCC MYA 4823 transformed with pCambia-Nou-PAP1 , 95.7 ± 3.2 , 1.1 ± 0.4 , This work.

Techniques: Generated, Staining, Spectrophotometry

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Cell wall rhamnose and mannose content in Sporothrix schenckii and Sporothrix brasiliensis wild-type, control, and PAP1 -silenced mutant strains. Yeast-like cells were homogenized, cell walls were isolated, cleansed, acid-hydrolyzed, and the resulting monosaccharides were separated and quantified by high-performance anion-exchange chromatography with pulsed amperometric detection. In A, results generated with S. schenckii cells. WT, 1099–18 ATCC MYA 4821 strain. In B, results obtained with S. brasiliensis strains. WT, 5110 ATCC MYA 4823 strain. For both panels, data are means ± SD of three biological replicates. Results were analyzed with Dunnett's test and then the t -test. * P < 0.05 when compared to WT, or control strains (HSS67 and HSS68 in panel A; HSB28 and HSB29 in panel B).

Article Snippet: HSB32 , 5110 ATCC MYA 4823 transformed with pCambia-Nou-PAP1 , 95.7 ± 3.2 , 1.1 ± 0.4 , This work.

Techniques: Control, Mutagenesis, Isolation, Chromatography, Generated

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Cytokine stimulation by human peripheral blood mononuclear cells. Yeast-like cells and human peripheral blood mononuclear cells were coincubated for 24 h, and the secreted cytokines were quantified by ELISA. Panels A , C , and E are results generated with S. schenckii yeast-like cells (WT, 1099–18 ATCC MYA 4821 strain), while panels B , D , and F are results obtained with S. brasiliensis strains (WT, 5110 ATCC MYA 4823 strain). None, human cells preincubated with 5 μg mL −1 polymyxin B; Anti-MR, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-mannose receptor antibody. Anti-TLR4, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-TLR4. Data are means ± SD obtained with samples from eight donors, each assayed in duplicate wells. Results were analyzed with Dunnett's test and then the Mann-Whitney U test. * P < 0.05 when compared to strains WT, HSS67, or HSS68 (panels A , C , and E ). * P < 0.05 when compared to strains WT, HSB28, or HSB29 (panels B , D , and F ). † P < 0.05 when compared to the “None” group from the same strain.

Article Snippet: HSB32 , 5110 ATCC MYA 4823 transformed with pCambia-Nou-PAP1 , 95.7 ± 3.2 , 1.1 ± 0.4 , This work.

Techniques: Enzyme-linked Immunosorbent Assay, Generated, MANN-WHITNEY

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Phagocytosis of Sporothrix schenckii and Sporothrix brasiliensis PAP1 -silenced strains. Yeast-like cells were labeled with Acridine Orange and used to interact with human monocyte-derived macrophages for 2 h at 37 °C and 5 % (v/v) CO 2 . Then, macrophages were collected and analyzed by flow cytometry. Macrophages that were interacting with at least one red fluorescent yeast-like cell were included in the analysis. None, human cells preincubated with 5 μg mL-1 polymyxin B. Anti-MR, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-mannose receptor antibody. Anti-TLR4, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-TLR4. Panel A , results generated with S. schenckii yeast-like cells (WT, 1099–18 ATCC MYA 4821 strain); while in panel B are results obtained with S. brasiliensis strains (WT, 5110 ATCC MYA 4823 strain). Data are means ± SD obtained with samples from eight donors, each assayed in duplicate wells. Results were analyzed with Dunnett's test and then the Mann-Whitney U test. * P < 0.05 when compared to strains WT, HSS67, or HSS68 (panel A ). * P < 0.05 when compared to strains WT, HSB28, or HSB29 (panel B ). † P < 0.05 when compared to the “None” group from the same strain. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: HSB32 , 5110 ATCC MYA 4823 transformed with pCambia-Nou-PAP1 , 95.7 ± 3.2 , 1.1 ± 0.4 , This work.

Techniques: Labeling, Derivative Assay, Flow Cytometry, Generated, MANN-WHITNEY

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Cytokine stimulation by human monocyte-derived macrophages. Yeast-like cells and monocyte-derived macrophages were coincubated for 24 h, and the secreted cytokines were quantified by ELISA. Panels A and C are results generated with S. schenckii yeast-like cells (WT, 1099–18 ATCC MYA 4821 strain); while panels B and D are results obtained with S. brasiliensis strains (WT, 5110 ATCC MYA 4823 strain). None, human cells preincubated with 5 μg mL −1 polymyxin B. Anti-MR, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-mannose receptor antibody. Anti-TLR4, human cells preincubated with 5 μg mL −1 polymyxin B and 10 μg mL −1 anti-TLR4. Data are means ± SD obtained with samples from eight donors, each assayed in duplicate wells. Results were analyzed with Dunnett's test and then the Mann-Whitney U test. * P < 0.05 when compared to strains WT, HSS67, or HSS68 (panels A and C ). * P < 0.05 when compared to strains WT, HSB28, or HSB29 (panels B and D ). cells. † P < 0.05 when compared to the “None” group from the same strain.

Article Snippet: HSB32 , 5110 ATCC MYA 4823 transformed with pCambia-Nou-PAP1 , 95.7 ± 3.2 , 1.1 ± 0.4 , This work.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Generated, MANN-WHITNEY

Journal: The Cell Surface

Article Title: Pap1 is an adhesin involved in the interaction of Sporothrix schenckii and Sporothrix brasiliensis with the host

doi: 10.1016/j.tcsw.2025.100164

Figure Lengend Snippet: Killing curves of Galleria mellonella larvae inoculated with Sporothrix schenckii and Sporothrix brasiliensis WT, control, and PAP1 -silenced strains. The G. mellonella larvae were inoculated with 10 μL of a yeast-like cell suspension at 1 × 10 7 cells mL -1, and survival was monitored daily for two weeks. Mortality is shown in Kaplan–Meier plots. In A, data generated with S. scehcnkii strains; while in B are shown the results with S. brasiliensis strains. Data were analyzed with the Log-rank test. In both panels, curves generated with the five PAP1 -silenced strains were not significantly different among themselves but were significantly different from the curves generated with the WT or control strains ( P < 0.05). In A, WT, 1099–18 ATCC MYA 4821 strain. Control strains are HSS67 and HSS68. In B, WT, 5110 ATCC MYA 4823 strain. Control strains are HSB28 and HSB29. For each strain, a total of 30 larvae were included in the study. In both panels, PBS, a larval group inoculated only with phosphate saline buffer.

Article Snippet: HSB32 , 5110 ATCC MYA 4823 transformed with pCambia-Nou-PAP1 , 95.7 ± 3.2 , 1.1 ± 0.4 , This work.

Techniques: Control, Suspension, Generated, Saline