Ations in response to oenothein B and IL-18 might be due to variable numbers of NK cells Title Loaded From File between PBMC samples or variable influences by other cells within the mixed populations on the NK cells, human NK cells were sorted, and then equal cell numbers were treated with oenothein B alone, IL-18 alone, or a combination of both. IFNc Vitamin D2 web production was measured 24 hrs later by ELISA. As shown in Figure 8, oenothein B alone directly induced IFNc production by NK cells and there was an increase in IFNc production with the combined treatment in all donors tested, although the amount of IFNc produced varied between donors. This variability in IFNc production by NK cells has been observed in other studies and may have a genetic component [42], [43]. These data further suggest that, as with bovine cells, oenothein B treatment has 23115181 the potential to augment IFNc production by human NK cells alone and in response to IL-18. In addition, thesedata suggest that oenothein B can directly prime these cells to respond to IL-18. Collectively, our results show that, in addition to monocytes, oenothein B stimulates subsets of bovine and human lymphocytes, including NK cells, CD8+ T cells, and both Vd2+ and Vd2- cd T cells, by upregulating IL-2Ra and/or CD69 on these cells. We also show that oenothein B promotes the production of IFNc by human lymphocytes, specifically cd T cells and CD8+ T cells. Furthermore, we demonstrate that IFNc production by NK cells can also be induced by oenothein B, although this response was not as robust or consistent as that seen in T cells. Interestingly, differences in the capacity of oenothein B to induce IFNc production by T cells was observed between human and bovine cells, as oenothein B alone did not directly induce significant IFNc secretion by bovine T cells as it did with human T cells. These data suggest that certain polyphenols may exert species-specific effects and that immunomodulatory effects of polyphenols demonstrated in one species may not always be conserved in other species. Thus, analysis of the immunomodulating properties of polyphenols cannot rely solely on animal testing, and a combination of animal and human cell testing is required to identify relevant, conserved responses. A possible explanation for some of the differences observed between human and bovine T cells in these studies could be due to differences in ages, as young calves 15857111 were used for our bovine studies while adults were used for our human studies. It has been shown that IFNc secretion by T cells can increase with age, correlating with an increase in CD45RO+ T cells [44]. Therefore, future studies could examine the effects of aging on these responses. It is possible that lymphocyte responses to certain polyphenols in young bovine calves are more reflective of those that might occur in children, suggesting a potential new use for this animal model in the study of the effects of dietary polyphenols on neonatal and adult lymphocytes. A potentially important and conserved response to oenothein B is enhanced IFNc secretion following exposure to suboptimal IL18 concentrations, which was observed in both bovine and human NK cells. The synergistic effect of oenothein B and IL-18 for enhancing IFNc production by NK cells was observed in mixed PBMC cultures, NK cell-depleted PBMCs, as well as sorted NK cells. Our earlier studies demonstrated that oenothein B couldStimulation of Lymphocytes by Oenothein Binduce IL-12 production by monocytes [7], which others.Ations in response to oenothein B and IL-18 might be due to variable numbers of NK cells between PBMC samples or variable influences by other cells within the mixed populations on the NK cells, human NK cells were sorted, and then equal cell numbers were treated with oenothein B alone, IL-18 alone, or a combination of both. IFNc production was measured 24 hrs later by ELISA. As shown in Figure 8, oenothein B alone directly induced IFNc production by NK cells and there was an increase in IFNc production with the combined treatment in all donors tested, although the amount of IFNc produced varied between donors. This variability in IFNc production by NK cells has been observed in other studies and may have a genetic component [42], [43]. These data further suggest that, as with bovine cells, oenothein B treatment has 23115181 the potential to augment IFNc production by human NK cells alone and in response to IL-18. In addition, thesedata suggest that oenothein B can directly prime these cells to respond to IL-18. Collectively, our results show that, in addition to monocytes, oenothein B stimulates subsets of bovine and human lymphocytes, including NK cells, CD8+ T cells, and both Vd2+ and Vd2- cd T cells, by upregulating IL-2Ra and/or CD69 on these cells. We also show that oenothein B promotes the production of IFNc by human lymphocytes, specifically cd T cells and CD8+ T cells. Furthermore, we demonstrate that IFNc production by NK cells can also be induced by oenothein B, although this response was not as robust or consistent as that seen in T cells. Interestingly, differences in the capacity of oenothein B to induce IFNc production by T cells was observed between human and bovine cells, as oenothein B alone did not directly induce significant IFNc secretion by bovine T cells as it did with human T cells. These data suggest that certain polyphenols may exert species-specific effects and that immunomodulatory effects of polyphenols demonstrated in one species may not always be conserved in other species. Thus, analysis of the immunomodulating properties of polyphenols cannot rely solely on animal testing, and a combination of animal and human cell testing is required to identify relevant, conserved responses. A possible explanation for some of the differences observed between human and bovine T cells in these studies could be due to differences in ages, as young calves 15857111 were used for our bovine studies while adults were used for our human studies. It has been shown that IFNc secretion by T cells can increase with age, correlating with an increase in CD45RO+ T cells [44]. Therefore, future studies could examine the effects of aging on these responses. It is possible that lymphocyte responses to certain polyphenols in young bovine calves are more reflective of those that might occur in children, suggesting a potential new use for this animal model in the study of the effects of dietary polyphenols on neonatal and adult lymphocytes. A potentially important and conserved response to oenothein B is enhanced IFNc secretion following exposure to suboptimal IL18 concentrations, which was observed in both bovine and human NK cells. The synergistic effect of oenothein B and IL-18 for enhancing IFNc production by NK cells was observed in mixed PBMC cultures, NK cell-depleted PBMCs, as well as sorted NK cells. Our earlier studies demonstrated that oenothein B couldStimulation of Lymphocytes by Oenothein Binduce IL-12 production by monocytes [7], which others.
