Monoclonal antibody based ELISAs for measurement of activins in biological fluids

Wong, Wai Lee; Garg, Shaily; Woodruff, Teresa K.; Bald, Laura; Fendly, Brian M.; Lofgren, James A.

doi:10.1016/0022-1759(93)90100-l

Cited by 39 publications

(29 citation statements)

References 37 publications

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“…01% cross-reactivity with related molecules. 19 Thus, the results of the present studies, based on this specific ELISA assay and the bioassay, indicate that the functional, dimeric activin A molecule is produced by monocytes.…”

Section: Discussionmentioning

confidence: 81%

Induced expression of the new cytokine, activin A, in human monocytes: inhibition by glucocorticoids and retinoic acid

Shao

Frigon

et al. 1996

Immunology

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The capacity of recombinant human granulocyte–macrophage colony‐stimulating factor (GM‐CSF), glucocorticoids or all‐trans‐retinoic acid to modulate production of activin A by human monocytes was studied. It was shown that GM‐CSF stimulated monocytes to accumulate activin A RNA after as few as 4 hr of incubation, reaching a peak of stimulation at approximately 16 hr of incubation. The activin A transcripts accumulated in the monocytes after stimulation with only 5 U/ml of GM‐CSF and reached a maximum plateau level of expression between 25 and 50 U/ml of GM‐CSF. Biologically active activin A molecules were detected in the conditioned media by a bioassay, performed both in the absence and presence of a neutralizing antiserum for activin A. Accumulation of bioactive activin A in conditioned medium of monocyte cultures was detected after 24 hr of incubation with GM‐CSF and high levels of activin A were maintained for 72 hr. The production of the dimeric βAβA in these monocytes was further confirmed by a sandwich enzyme‐linked immunosorbent assay (ELISA) specific for activin A. In contrast to the stimulatory effect of GM‐CSF, hydrocortisone, dexamethasone or all‐trans‐retinoic acid at 1 × 10−7 to 1 × 10−5 m inhibited the constitutive expression of activin A and greatly suppressed the GM‐CSF‐stimulated production. Thus, the expression of activin A is modulated in monocytes by different agents. These observations may imply new roles for activin A at sites of inflammation where monocytes accumulate.

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Section: Discussionmentioning

confidence: 81%

Induced expression of the new cytokine, activin A, in human monocytes: inhibition by glucocorticoids and retinoic acid

Shao

Frigon

et al. 1996

Immunology

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“…By using an ELISA (19), the levels of activin A (l3A:4A homodimer) in the serum of these mice were found to be elevated 13-fold (males) and 20-fold (females) ( Table 1). Similar elevations of serum activin B (,BB:,BB homodimers) were also observed (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…Serum was frozen at -200C before analysis. Serum activin A and B levels were determined by an ELISA method as described (19).…”

Section: Methodsmentioning

confidence: 99%

Development of cancer cachexia-like syndrome and adrenal tumors in inhibin-deficient mice.

Matzuk

Finegold

Mather

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

343

266

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Activins and inhibins, members of the tWe ( transforming growth factor superfamily of growth regulatory proteins, are produced in multiple tissues and affect diverse physiologic processes. Using embryonic stem cell technology, we previously demonstrated that inhibin can function as a gonadal tumor suppressor. In this study, we show that development of gonadal tumors is rapidly followed by a cancer cachexia-like wasting syndrome. Cachectic inhibin-deficient mice develop hepatocellular necrosis around the central vein and parietal cell depletion and mucosal atrophy in the glandular stomach, are anemic, and demonstrate severe weight loss. The liver pathology is consistent with studies demonstrating an effect of elevated activins on rat hepatocytes. In inhibindeficient mice with tumors, activins are >10-fold elevated in the serum and are likely causing some of the cachexia symptoms. In contrast, inhibin-deficient mice gonadectomized at an early age do not develop this wasting syndrome. However, these gonadectomized, inhibin-deficient mice eventually develop adrenal cortical sex steroidogenic tumors with nearly 100% penetrance, demonstrating that inhibin is also a tumor suppressor for the adrenal gland.The inhibins (a:(3 heterodimers) and activins (13:l3dimers) are related proteins that share common 13 subunits (either 13A or PB subunits) (1). These proteins, members of a large family of structurally related growth regulatory proteins, which includes the type (3 transforming growth factors (TGF-P), bone morphogenetic proteins, and Mullerian inhibiting substance (MIS) (1-4), have been shown to have diverse functions in a variety of assay systems including antiproliferative effects on carcinoma cell lines [i.e., MIS and TGF-p (2, 3)].The activins and inhibins also have diverse functions in mammalian physiology and development. Activin and inhibin subunit mRNAs and proteins are synthesized in a variety of cell types and embryonic and adult tissues in different species (1,(5)(6)(7)(8)(9)(10). In the adult mammal, although activins were initially discovered for their ability to stimulate pituitary follicle-stimulating hormone secretion, they have also been shown to influence other functions including liver metabolism and glucose regulation. For example, activin A can stimulate glycogenolysis (11) and inhibit DNA synthesis in isolated rat hepatocytes in vitro (12). In addition, infusion of activin A into rats caused hepatocellular necrosis around the central vein of the liver (13). These physiologic effects of activin on hepatocytes are consistent with findings that a major site of 125I-labeled activin A binding is the rat liver (14) and that the type II activin receptor is expressed in the mouse liver (ref. 15; R. Towns and M.M.M., unpublished data).Using embryonic stem cell technology, we have generated inhibin-deficient mice (16), which develop sex cord stromal tumors at an early age with nearly 1001% penetrance, demonstrating that inhibin functions in vivo as a tumor suppressor in the gonads of mice (16). In th...

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“…Recombinant human inhibin A was generated in the laboratory. Antibodies that recognize FS, activin, and inhibin used in this study have been previously described [15,16]. Normally cycling and timed-pregnant Sprague-Dawley-derived female rats were obtained from Harlan BioSciences (Indianapolis, IN).…”

Section: Reagents and Sample Collectionmentioning

confidence: 99%

Identification of Naturally Occurring Follistatin Complexes in Human Biological Fluids1

Wang¹,

Draper²,

Lee³

et al. 1999

Biology of Reproduction

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Follistatin (FS) binds activin and inhibin proteins. Many organs are sensitive to activin and inhibin; thus the formation of FS-activin/inhibin complexes is important to our understanding of ligand activity. Other investigators studying FS have detected large molecular weight immunoreactive FS bands (greater than the expected molecular weight of FS alone) that have not been well characterized. The goal of this study was to identify naturally occurring FS monomers and FS-activin/inhibin complexes in several organ systems. The pituitary, ovary, kidney, and urine were chosen for this investigation. Molecular masses were assigned to in vitro assemblies of complexes containing recombinant inhibin or activin with FS for comparison with naturally occurring FS forms. The recombinant complex of FS-activin was primarily 97-kDa size, while FS-inhibin complexes were detected in a range of molecular sizes from 66 kDa to 97 kDa, 133 kDa, and > 220 kDa. FS-containing complexes of 66-kDa, 97-kDa, and 133-kDa were identified in the tissues examined and in pregnant urine. Our study points to the assembly of a series of FS-activin/inhibin complexes in a variety of organ systems that may impact upon the available amount of free versus bound (or "complexed") ligand, which must be considered when investigating the biology of activin- or inhibin-responsive cells. In addition, urine may be an important biological fluid that can be used to measure significant changes in circulating FS complexes.

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Monoclonal antibody based ELISAs for measurement of activins in biological fluids

Cited by 39 publications

References 37 publications

Induced expression of the new cytokine, activin A, in human monocytes: inhibition by glucocorticoids and retinoic acid

Induced expression of the new cytokine, activin A, in human monocytes: inhibition by glucocorticoids and retinoic acid

Development of cancer cachexia-like syndrome and adrenal tumors in inhibin-deficient mice.

Identification of Naturally Occurring Follistatin Complexes in Human Biological Fluids1

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