Ch lectin to interact with epithelial cell's receptors, labeled HHA and GNA were incubated with epithelial cells. As depicted in Figure 2C, only HHA but not GNA interacted with receptors expressed at the surface of HEC-1. Moreover, the preincubation of GNA with epithelial cells followed with several washes did not inhibit viral attachment whereas the preincubation of GNA with R5 viral particles in
Achment on HEC-1A Considering that both HHA and GNA were non-toxic in the epithelial cell system, we further investigated the inhibition of HIV adsorption on the apical side of HEC-1A by the compounds. Cells were thus incubated with HIV-1Ba-L in the presence or absence of different concentrations of HHA or GNA. As depicted in Figure 2A, HHA faintly decreased the attachment of HIV-1Ba-L on epitheli
Us combinations of plasmids. Bar, 10 m. In the absence of a viral RNA, MS2-YFP had the appearance shown in Fig. 4A to C. (A) pFIVgag-CFP, pMS2-YFP, and pFIV -24. The nuclear envelope region (box, right panel) is shown enlarged below (scale bar, 5 m). (B) pFIVgag-CFP, pFP93, pMS2-YFP, and pFIV -24. (C and D) pFIVgag-CFP, pFP93, pMS2-YFP, and pFIV -24. Live-cell confocal imaging was performed 48 h a
Us combinations of plasmids. Bar, 10 m. In the absence of a viral RNA, MS2-YFP had the appearance shown in Fig. 4A to C. (A) pFIVgag-CFP, pMS2-YFP, and pFIV -24. The nuclear envelope region (box, right panel) is shown enlarged below (scale bar, 5 m). (B) pFIVgag-CFP, pFP93, pMS2-YFP, and pFIV -24. (C and D) pFIVgag-CFP, pFP93, pMS2-YFP, and pFIV -24. Live-cell confocal imaging was performed 48 h a
Ation MDDC are washed and 2 ?105 cells were adsorbed on a microscopy-adapted slide. Cells were incubated with or without HHA-TRITC or GNA-TRITC (200 /ml; Eylabs) at 4 for 30 min. Cells were washed with PBS 0.01 azide 0.5 BSA and then fixed with 1 paraformaldehyde. The coverslides were mounted in Mowiol (SigmaAldrich). Fluorescence analysis was performed with a Zeiss LSM510 confocal microscope
Ation MDDC are washed and 2 ?105 cells were adsorbed on a microscopy-adapted slide. Cells were incubated with or without HHA-TRITC or GNA-TRITC (200 /ml; Eylabs) at 4 for 30 min. Cells were washed with PBS 0.01 azide 0.5 BSA and then fixed with 1 paraformaldehyde. The coverslides were mounted in Mowiol (SigmaAldrich). Fluorescence analysis was performed with a Zeiss LSM510 confocal microscope
Tration of the study plant lectins in epithelial cell (A). MDM)line (HEC-1A) and primary immune cells (MDDC (A). Non-toxic concentration of the study plant lectins in epithelial cell line (HEC-1A) and primary immune cells (MDDC and MDM). HEC-1A and primary immune cells were cultured with concentrations of products for 24 h. After washing, culture viability was determined by using the MTT-cytotoxic
S) and cells were lysed (1 Triton X-100 for 45 min at 37 ). Cell lysates were harvested and centrifuged at 1,800 rpm for 5 min. The amount of HIV-p24 antigen associated to cell lysates was determined using the HIV-1 p24 core profile ELISA. HIV-1 attachment on MDDC to assess the attachment of HIV-1 to MDDC, the cells were washed 2 times after 6 days of differentiation and seeded into 96-well cultu