Journal: Communications Biology

Article Title: Carbohydrate-mediated interactions between chloroviruses and the immune system

doi: 10.1038/s42003-024-07244-9

Figure Lengend Snippet: A Human C-type lectins ELISA results: wells were coated with viral glycoproteins and the binding to DC-SIGN, Langerin and MGL was evaluated in a calcium-containing buffer. B Plant lectins ELISA results: wells were coated with viral glycoproteins and the binding to VVL, HPA, SBA and RCA was evaluated. The experiments were performed in duplicate and data were normalized over signal from BSA-coated wells. Error bars indicate standard deviations. OD Optical density, VVL V icia villosa lectin, HPA Helix pomatia agglutinin, SBA Soybean agglutinin, RCA Ricinus communis agglutinin.

Article Snippet: PBMCs were then loaded on a Percoll layer (GE 5 Healthcare, Chicago, U.S.) and after centrifugation, the interphase was collected, washed, and resuspended in complete RPMI medium. moDCs were generated by culturing monocytes for 5–7 days at a concentration of 1.25 × 10 6 /mL in RPMI medium containing 500 U/mL IL-4 (ImmunoTools, Friesoythe, Germany) and 800 U/mL Granulocyte Macrophage Colony-stimulating Factor (GM-CSF) (ImmunoTools). moDCs (0.5–1 × 10 5 ) at day 4 after the monocytes stimulation were challenged with 20 μg/mL of different glycosylated MCPs in RPMI-1640, supplemented with 10% foetal calf serum, L-glutamine (2 mM), and penicillin/streptomycin (100 U/mL) at 37 °C, 5% CO 2 for 16 h. Human IL-10 and IL-6 ELISA Kit Duo Set R&D systems were used following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: Communications Biology

Article Title: Carbohydrate-mediated interactions between chloroviruses and the immune system

doi: 10.1038/s42003-024-07244-9

Figure Lengend Snippet: IL-6 ( A ) and IL-10 ( B ) secretion from human monocytes-derived dendritic cells ( n = 4) stimulated with the viral glycoproteins at 20 μg/mL, detected via ELISA in the supernatants after 20 h stimulation. Mean values from four different experiments (four different human donors) are reported. Error bars indicate standard deviations. Experimental data were analysed by ANOVA multiple comparison (Tukey’s multiple comparison test), with a statistical significance level of alpha=0.05. Significance was defined as * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: PBMCs were then loaded on a Percoll layer (GE 5 Healthcare, Chicago, U.S.) and after centrifugation, the interphase was collected, washed, and resuspended in complete RPMI medium. moDCs were generated by culturing monocytes for 5–7 days at a concentration of 1.25 × 10 6 /mL in RPMI medium containing 500 U/mL IL-4 (ImmunoTools, Friesoythe, Germany) and 800 U/mL Granulocyte Macrophage Colony-stimulating Factor (GM-CSF) (ImmunoTools). moDCs (0.5–1 × 10 5 ) at day 4 after the monocytes stimulation were challenged with 20 μg/mL of different glycosylated MCPs in RPMI-1640, supplemented with 10% foetal calf serum, L-glutamine (2 mM), and penicillin/streptomycin (100 U/mL) at 37 °C, 5% CO 2 for 16 h. Human IL-10 and IL-6 ELISA Kit Duo Set R&D systems were used following the manufacturer’s instructions.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Communications Biology

Article Title: Carbohydrate-mediated interactions between chloroviruses and the immune system

doi: 10.1038/s42003-024-07244-9

Figure Lengend Snippet: Concentration of IL-6 ( A ) and IL-10 ( B ) secreted from RAW264.7 Lucia macrophage cell line challenged with 20 µg/mL of the MCP glycans of PBCV-1, ATCV-1, OSy-NE5, P91, EIL-1, and PIL6. Accumulated IL-6 and IL-10 concentrations in 20 h supernatants were measured with ELISA. Data are means ± standard error ( n = 4). Experimental data were analysed by ANOVA multiple comparison (Tukey’s multiple comparison test), with a statistical significance level of alpha=0.05. Significance was defined as * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: PBMCs were then loaded on a Percoll layer (GE 5 Healthcare, Chicago, U.S.) and after centrifugation, the interphase was collected, washed, and resuspended in complete RPMI medium. moDCs were generated by culturing monocytes for 5–7 days at a concentration of 1.25 × 10 6 /mL in RPMI medium containing 500 U/mL IL-4 (ImmunoTools, Friesoythe, Germany) and 800 U/mL Granulocyte Macrophage Colony-stimulating Factor (GM-CSF) (ImmunoTools). moDCs (0.5–1 × 10 5 ) at day 4 after the monocytes stimulation were challenged with 20 μg/mL of different glycosylated MCPs in RPMI-1640, supplemented with 10% foetal calf serum, L-glutamine (2 mM), and penicillin/streptomycin (100 U/mL) at 37 °C, 5% CO 2 for 16 h. Human IL-10 and IL-6 ELISA Kit Duo Set R&D systems were used following the manufacturer’s instructions.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Communications Biology

Article Title: Carbohydrate-mediated interactions between chloroviruses and the immune system

doi: 10.1038/s42003-024-07244-9

Figure Lengend Snippet: A IgG ELISA results: wells were coated with viral glycoproteins and human IgG was specifically detected. Single OD normalized values are reported from 12 different donors, as well as the mean value from each group. Data were normalized over signal from BSA-coated wells. B Heat map from the IgG ELISA results. OD: Optical density. Experimental data were analysed by ANOVA multiple comparison (Tukey’s multiple comparison test), with a statistical significance level of alpha=0.05. Significance was defined as * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: PBMCs were then loaded on a Percoll layer (GE 5 Healthcare, Chicago, U.S.) and after centrifugation, the interphase was collected, washed, and resuspended in complete RPMI medium. moDCs were generated by culturing monocytes for 5–7 days at a concentration of 1.25 × 10 6 /mL in RPMI medium containing 500 U/mL IL-4 (ImmunoTools, Friesoythe, Germany) and 800 U/mL Granulocyte Macrophage Colony-stimulating Factor (GM-CSF) (ImmunoTools). moDCs (0.5–1 × 10 5 ) at day 4 after the monocytes stimulation were challenged with 20 μg/mL of different glycosylated MCPs in RPMI-1640, supplemented with 10% foetal calf serum, L-glutamine (2 mM), and penicillin/streptomycin (100 U/mL) at 37 °C, 5% CO 2 for 16 h. Human IL-10 and IL-6 ELISA Kit Duo Set R&D systems were used following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison