lactoferrin and resazurin

Two bovine mammary cell types (BME-UV1 and MeBo cells) were used to evaluate the effect of natural retinoids, retinoid analogs, and bovine lactoferrin (bLf) on cell viability in vitro. Experiments with Alamar Blue showed a linear relationship between fluorescence and cell viability index. The BME-UV1 cells exhibited twice the metabolic activity but required half the doubling time of the MeBo cells. The BME-UV1 cells were very sensitive to all-trans retinoic acid (atRA) inhibition of cell viability (P<0.05) and exhibited a dose-dependent inhibition with 9-cisRA (9cRA; P<0.05). The MeBo cells exhibited some inhibition with these natural ligands (P<0.05), but they were not as sensitive. The addition of bLf had similar inhibitory effects (P<0.05) on cell viability of the 2 mammary cell types. Applications of RA receptor (RAR) agonist indicated that the stimulation of the RAR in both mammary cell types was highly effective in inhibition of cell viability (P<0.05), whereas the application of an RAR antagonist stimulated MeBo cell viability (P<0.05) and inhibited BME-UV1 cell viability (P<0.05). Finally, the use of the RAR antagonist in conjunction with bLf indicated a rescue of the bLf effect in the MeBo cells, suggesting that bLf is acting through the RAR receptor. Conversely, bLf reverted inhibition of cell viability by 9cRA in the BME-UV1 cell type (P<0.05). We conclude that RAR interaction in bovine mammary cell types regulates cell viability in vitro; we hypothesize that the natural ligands mediate regulation of bovine mammary cell viability in vivo and that bLf can either enhance or reverse the retinoid-induced inhibition of cell viability, depending on the type of bovine mammary cell studied.

Topics: Animals; Cattle; Cell Line; Cell Survival; Drug Interactions; Epithelial Cells; Female; Indicators and Reagents; Lactoferrin; Mammary Glands, Animal; Oxazines; Receptors, Retinoic Acid; Retinoids; Tretinoin; Xanthenes

Human serum has been reported to inhibit the growth of several fungal pathogens. Serum inhibitory activity has predominantly been associated with transferrin-mediated iron binding. We found that serum from several animal species (human, dog, mouse and fetal bovine) inhibited the growth of the dimorphic fungal pathogen, Blastomyces dermatitidisin vitro. In initial studies, we found no correlation between the total iron-binding capacity of various sera and their inhibitory activity against B. dermatitidis yeast. In addition, we were unable to abrogate the inhibitory activity of human serum against B. dermatitidis yeast by the addition of ferric chloride (10-200 microM). We found that apo-transferrin had little or no inhibitory activity against B. dermatitidis yeast. Furthermore, serum from hypotransferrinemic mice (hpx/hpx) had inhibitory activity against B. dermatitidis yeast that was equivalent to that of normal mouse serum. These findings are consistent with our initial findings and suggest that serum inhibitory activity against B. dermatitidis yeast is transferrin independent. Although high concentrations (3.5-5 mg/ml) of lactoferrin did have inhibitory activity against B. dermatitidis yeast, these concentrations were orders of magnitude greater than those found in serum under normal physiological conditions, suggesting that lactoferrin was not likely to contribute to serum inhibitory activity against B. dermatitidis yeast. Our findings suggest that host iron-binding proteins may be relatively ineffective in innate host defense against infection with B. dermatitidis yeast.

Topics: Animals; Blastomyces; Blood Physiological Phenomena; Cattle; Chlorides; Coloring Agents; Dogs; Ferric Compounds; Humans; Lactoferrin; Mice; Mice, Inbred ICR; Oxazines; Transferrin; Xanthenes