Yukiko Tokuyama

Bovine colostric transforming growth factor-β-like peptide that induces growth inhibition and changes in morphology of human osteogenic sarcoma cells(MG-63)

TGF-beta like peptide, termed TGF(BC-1), was partially purified from defatted and decaseinated bovine colostrum by a sequence of DEAE-Sephacel chromatography and Sephadex G-50 gel filtration in 1M acetic acid. TGF(BC-1) was distinct from well-known 25K TGF-beta in chemical properties: TGF(BC-1) was sensitive to acid ethanol extraction (Roberts et al., 1980). Its apparent molecular weight ranged from 21k to 11k by gel filtration and it was composed of low MW peptides (15k, 13k, 10k and 7.3k but not 25k) as examined by SDS-PAGE under non-reducing conditions. However, TGF(BC-1) shares some biological properties with the prototype TGF-b. TGF(BC-1) remarkably suppressed growth of osteogenic sarcoma cells (MG-63), and this was intriguingly accompanied by a striking change in morphology.

Purification and identification of TGF-β2-related growth factor from bovine colostrum

Bovine colostrum contains transforming growth factor (TGF)-beta-like activity. High levels of this activity are found in early colostrum (within 12 h after parturition); however, it decreases rapidly after 30 h. In this study, the activity in early colostrum was purified by a sequence of decaseination, DEAE-Sephacel chromatography, Sephadex G-100 gel filtration, precipitation at neutral pH, further gel filtration and reversed-phase FPLC. A homodimeric 23 kDa protein was identified that retained the TGF-beta-like activity. Its activity was completely neutralized by a specific anti-TGF-beta 2 antibody. These results indicate that the TGF-beta-like growth factor found in bovine colostrum is chemically and antigenically related to TGF-beta 2. The physiological roles of the TGF-beta 2-related growth factor are discussed.

Retinoic acid and steroid hormones regulate IgA production by LPS-stimulated murine spleen cells

We examined the regulatory effects of steroid hormones (beta-estradiol and glucocorticoids) on in vitro IgA production. Addition of retinoic acid (RA, 100-500 nM) to the LPS-stimulated spleen cell culture enhanced IgA production (8-22-fold). Simultaneous addition of beta-estradiol, but not testosterone, enhanced the effect of RA synergistically (a further 2-4-fold). In contrast, glucocorticoids inhibited the reaction. The concentration inhibiting IgA production by 50% was 1 nM, 6 nM and 10 nM for dexamethasone, prednisolone and hydrocortisone, respectively. None of the hormones tested alone affected IgA production by LPS-stimulated spleen cells. Hydrocortisone enhanced the IgG1 production by LPS-stimulated spleen cells. This effect was completely abolished by simultaneous addition of RA. These findings indicate that RA can direct the class-switching to IgA in LPS-stimulated spleen cells, and that beta-estradiol and glucocorticoids have positive and negative regulatory effects, respectively, on the IgA production.