Compared with those of standard OD490 versus cell number curves generated for each cell type. The percentage survival was calculated using the formula :Epithelial monolayer integrity the effect of the plant lectins on their ability to maintain an intact epithelium was determined by measuring transepithelial resistance (TER). HEC-1A cells were grown as a tight polarized monolayer on a permeable sup
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
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
L side of the transwell, and epithelial integrity was measured at 30 min and 2, 4, 9, and 24 h by using the Minicell-ERS instrument. The data shown are the averages from duplicate wells ?1 standard error of the mean after product addition.Page 4 of(page number not for citation purposes)Journal of Translational Medicine 2007, 5:http://www.translational-medicine.com/content/5/1/p24 (pg/ml)of the pla
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
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
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
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