Competitive protein binding assay for activin AEDF using follistatin determination of activin levels in human plasma

Demura, Reiko; Suzuki, Tomoharu; Tajima, Saeko; Mitsuhashi, S.; Odagiri, Emi; Eto, Yuzuru; Sugino, Hiromu; Demura, Hiroshi

doi:10.1016/0006-291x(92)91746-d

Cited by 18 publications

(4 citation statements)

References 15 publications

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“…FS and activin are detectable in serum (Demura et al, 1992(Demura et al, , 1993Sakai et al, 1992;Klein et al, 1993;Gilfillan & Robertson, 1994;Khoury et al, 1995;Knight et al, 1996;Tilbrook et al, 1996;McFarlane et al, 1996;Wakatsuki et al, 1996;Sakamoto et al, 1996) and FS concentrations in serum increase with age (Wakatsuki et al, 1996). Evidence that pharmacological concentrations of both polypeptides can affect pituitary hormone secretion in vivo, might suggest that the peptides could have endocrine functions (Inouye et al, 1991;Carroll et al, 1991;DePaolo et al, 1991: Stouffer et al, 1993Woodruff et al, 1993;Meriggiola et al, 1994;Tilbrook et al, 1996).…”

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confidence: 99%

Serum follistatin concentrations are increased in patients with septicaemia

Michel

Shintani

Nau

1998

Clinical Endocrinology

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confidence: 99%

Serum follistatin concentrations are increased in patients with septicaemia

Michel

Shintani

Nau

1998

Clinical Endocrinology

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“…Previous activin immunoassays (Groóme 1991, Robertson et al 1992, bioassays (Sakai et al 1992, Sadatsuki et al 1993) and proteinbinding assays (Demura et al 1992(Demura et al , 1993 were compro¬ mised by the presence of activin-binding proteins, for example, follistatin and a2M (Shimonaka et al 1991, Schneyer et al 1992, Vaughan & Vale 1993). However, we recently described a new two-site ELISA for total activin-A (Knight et al 1996) which was sensitive (detection limit 100 pg/ml), specific (there was no significant cross-reaction with any of the related molecules such as inhibin-A, inhibin-B, activin-B, follistatin and bovine pro-aC), accurate (activin-A recovery values of 102% in bFF and 96% in hFF) and not susceptible to interference by follistatin or a2M.…”

Section: Discussionmentioning

confidence: 95%

Development, validation and application of a two-site enzyme-linked immunosorbent assay for activin-AB

Evans

Muttukrishna²,

Knight³

et al. 1997

Journal of Endocrinology

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Monoclonal antibodies, specific for the beta A and beta B subunits of activin, were used to develop a new two-site ELISA for activin-AB. The assay had a detection limit of 0.19 ng/ml. High concentrations of activin-AB were found in bovine, ovine and porcine follicular fluids (FF), with less in human FF (1310, 1730, 688 and 7 ng/ml respectively). Recovery of spiked activin-AB standard from human, bovine and ovine FFs and from homogenized human placental extracts averaged 91%, 115%, 115% and 94% respectively. Within-plate coefficients of variation for different concentration of activin-AB were between 1.3% and 2.67%. The between-plate coefficient of variation was 5.5%. Cross-reactivity experiments showed the high specificity of the assay for activin-AB, with inhibin-A, inhibin-B, follistatin, activin-A and activin-B all cross-reacting < 0.2%. Incubation with high concentrations of follistatin (500 ng/ml) prior to assay did not affect the recovery of activin-AB. Samples of bovine, porcine, ovine and human FF gave dose responses parallel to that of the standard, as did bovine granulosa cell-conditioned media. In human and porcine FF, levels of activin-A and activin-AB were similar whereas, in bovine and ovine FF, activin-A levels were approximately threefold higher than activin-A, nearly all of the endogenous activin-AB in bovine FF was detected in the eluate from gel permeation chromatography with an M(r) of > 700000 indicating its association with higher molecular weight binding protein(s). By contrast, after denaturation, immunoreactive activin-AB was detected with an M(r) of approximately 25000 consistent with the complete dissociation from binding proteins. Activin-A was detected in relatively high concentrations in human FF (approximately 5 ng/ml), homogenized placental extracts (4.35-95.5 ng/g), sera from pregnant women (> 4 ng/ml) and amniotic fluid (3-13 ng/ml), and in much lower concentrations in postmenopausal serum (500 pg/ ml), normal cycle serum (100-200 pg/ml), serum from gonadotrophin-treated women (200 pg/ml), and normal adult male serum (225 pg/ml). Activin-A was also found in the culture media from explants of human amnion, chorion, maternal decidua and placenta. In marked contrast, activin-AB was undetectable (< 0.19 ng/ml) in all of these samples with the exception of human FF (approximately 7 ng/ml). In conclusion, we have developed a sensitive and specific ELISA to measure total (bound+free) activin-AB. Preliminary results show a more restricted distribution of this isoform compared with activin-A. The presence of high levels of both activin-A and activin-AB in FF suggests a function for both isoforms in the developing ovarian follicle.

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“…Localized production of cytokines that have mitogenic or chemotactic effects on SMCs produced by macrophages may be involved in the atherogenic process. 10 Alternatively, activin that is detectable in human plasma 34 may affect vascular cells via a hormonal mechanism.…”

Section: Discussionmentioning

confidence: 99%

Expression of follistatin, an activin-binding protein, in vascular smooth muscle cells and arteriosclerotic lesions.

Inoue

Orimo

Hosoi

et al. 1993

Arterioscler Thromb

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Activin-A has a mitogenic effect on vascular smooth muscle cells (SMCs) and is produced by monocyte/macrophage lineage cells. Here, we studied the expression of follistatin, an activin-binding protein, in both A10 cells, a rat aortic SMC line, and vascular SMCs derived from adult rat aorta. Follistatin mRNA was detected in these cells by Northern blot analysis. Ligand blot and immunoblot analyses demonstrated that follistatin was produced in the conditioned medium at a higher level by A10 cells than by SMCs. Furthermore, immunostaining and in situ hybridization of the arteriosclerotic lesions showed that follistatin was highly expressed in the diseased artery, where abnormal proliferation of SMCs occurred. We suggest that follistatin is produced by vascular SMCs and is involved in the course of atherogenesis. (Arterioscler Thromb. 1993;13:1859-1864 KEY WORDS • follistatin • activin • arteriosclerosis • smooth muscle cells S mooth muscle cell (SMC) proliferation in vivo plays a central role in the pathogenesis of atherosclerosis.

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Competitive protein binding assay for activin AEDF using follistatin determination of activin levels in human plasma

Cited by 18 publications

References 15 publications

Serum follistatin concentrations are increased in patients with septicaemia

Serum follistatin concentrations are increased in patients with septicaemia

Development, validation and application of a two-site enzyme-linked immunosorbent assay for activin-AB

Expression of follistatin, an activin-binding protein, in vascular smooth muscle cells and arteriosclerotic lesions.

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