Of plant lectins on HIV-1 transcytosis through a tight epithelial cells monolayer To evaluate the capacity of each plant lectin to inhibit in vitro HIV-1 transcytosis through genital epithelial cells, such as HEC-1A cells, we used a dual-chamber model in which the apical chamber consisted of a confluent monolayer of HEC-1A cells, and the basal chamber contained fresh medium. Cell-free virus (HIV-1
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
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
Vities in the epithelial cell line HEC-1A (i) Toxicity Any topical microbicide approved for human use first need to be evaluated for epithelial toxicity, because of the direct contact between the product and the vaginal mucosal surface. Thus, non-toxic concentrations of the plant lectins were determined by culturing the epithelial cell line, HEC-1A, for 24 h in the presence of different lectin con
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
Glycans (GAG) heparan sulfates, we assessed the role of plant lectins on the attachment of X4-tropic viruses, known to interact very efficiently with heparan sulfate [11] and data not shown]. Thus, cells were incubated with HIV-1NDK (Fig. 2B) in the presence or absence of different concentrations of HHA and GNA. As depicted in Figure 2B, none of the plant lectins, whatever the concentration, inhib
Glycans (GAG) heparan sulfates, we assessed the role of plant lectins on the attachment of X4-tropic viruses, known to interact very efficiently with heparan sulfate [11] and data not shown]. Thus, cells were incubated with HIV-1NDK (Fig. 2B) in the presence or absence of different concentrations of HHA and GNA. As depicted in Figure 2B, none of the plant lectins, whatever the concentration, inhib