Characterization of a low molecular mass form of insulin-like growth factor binding protein-3 (17.7 kilodaltons) in urine and serum from healthy children and growth hormone (GH)-deficient patients

Relationship with GH therapy

Anna Spagnoli, Sharron E. Gargosky, Gian Luigi Spadoni, Margaret MacGillivray, Youngman Oh, Brunetto Boscherini, Ron G. Rosenfeld

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The insulin-like growth factor binding proteins (IGFBPs) are the carriers for insulin-like growth factor (IGF0-I and IGF-II. IGFBP-3 is GH-dependent and circulates associated with IGFs and an acid-labile subunit to form a 150-kilodalton (kDa) complex. In human serum, two immunoreactive molecular weight forms of IGFBP-3 have been identified. In human urine, radioimmunoassayable levels of IGFBP-3 have been detected. The objectives of this study were to characterize the molecular weight forms of IGFBP-3 in urine and serum of healthy children and adults and in children with GH deficiency (GHD), to quantify the urinary molecular weight forms of IGFBP-3, and to evaluate the relationship of these forms with GH therapy. Urine and serum were obtained from 12 prepubertal children with GHD, before and after 6 months of GH therapy, from 30 prepubertal healthy children, and from 8 healthy adults. Western immunoblotting (WIB) with IGFBP-3 antiserum (alpha IG-FBP-3g1) showed that in urine the most representative IGFBP-3 was a 17.7-kDa form. The 17.7-kDa IGFBP-3 was high in urine of healthy children compared with healthy adults and was low in children with GHD but increased after GH therapy. Urinary IGFBP-3 immunoreactive profile was determined by neutral-size exclusion chromatography, followed by IGFBP-3 RIA analysis of the fractions. Urine showed a major peak of IGFBP-3 immunoreactivity around 17 kDa. The 17-kDa urinary IGFBP-3 chromatographic peak averaged 8461 ± 367 ng/12 h.m2 of body surface in healthy children, 3415 ± 739 in adults (P < 0.001), 2294 ± 354 in children with GHD before GH therapy (P < 0.001), and 7940 ± 1874 in children with GHD after GH therapy. Urinary IGFBP-3 was also measured by RIA in unfractionated urine; healthy children showed levels significantly higher (14575 ± 460 ng/12 h.m2) than adults (7823 ± 1083, P < 0.001) and higher than children with GHD before GH therapy (4710 ± 703, P < 0.001). Again, however, immunoreactive IGFBP-3 increased after GH treatment (12294 ± 3394). In the serum of the healthy children we characterized by specific IGFBP-3 WIB analysis, a 17.7-kDa immunoreactive form of IGFBP-3 that was absent in the serum of healthy adults and low in patients with GHD, increased during GH therapy. Serum samples were subjected to neutral-size exclusion chromatography and the fractions were analyzed by WIB.(ABSTRACT TRUNCATED AT 400 WORDS).

Original languageEnglish (US)
Pages (from-to)3668-3676
Number of pages9
JournalJournal of Clinical Endocrinology and Metabolism
Volume80
Issue number12
DOIs
StatePublished - Jan 1 1995

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Insulin-Like Growth Factor Binding Protein 3
Molecular mass
Growth Hormone
Urine
Serum
Therapeutics
Size exclusion chromatography
Molecular Weight
Western Blotting
Molecular weight
Gel Chromatography
Insulin-Like Growth Factor Binding Proteins
Insulin-Like Growth Factor II
Insulin-Like Growth Factor I

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Characterization of a low molecular mass form of insulin-like growth factor binding protein-3 (17.7 kilodaltons) in urine and serum from healthy children and growth hormone (GH)-deficient patients : Relationship with GH therapy. / Spagnoli, Anna; Gargosky, Sharron E.; Spadoni, Gian Luigi; MacGillivray, Margaret; Oh, Youngman; Boscherini, Brunetto; Rosenfeld, Ron G.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 80, No. 12, 01.01.1995, p. 3668-3676.

Research output: Contribution to journalArticle

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abstract = "The insulin-like growth factor binding proteins (IGFBPs) are the carriers for insulin-like growth factor (IGF0-I and IGF-II. IGFBP-3 is GH-dependent and circulates associated with IGFs and an acid-labile subunit to form a 150-kilodalton (kDa) complex. In human serum, two immunoreactive molecular weight forms of IGFBP-3 have been identified. In human urine, radioimmunoassayable levels of IGFBP-3 have been detected. The objectives of this study were to characterize the molecular weight forms of IGFBP-3 in urine and serum of healthy children and adults and in children with GH deficiency (GHD), to quantify the urinary molecular weight forms of IGFBP-3, and to evaluate the relationship of these forms with GH therapy. Urine and serum were obtained from 12 prepubertal children with GHD, before and after 6 months of GH therapy, from 30 prepubertal healthy children, and from 8 healthy adults. Western immunoblotting (WIB) with IGFBP-3 antiserum (alpha IG-FBP-3g1) showed that in urine the most representative IGFBP-3 was a 17.7-kDa form. The 17.7-kDa IGFBP-3 was high in urine of healthy children compared with healthy adults and was low in children with GHD but increased after GH therapy. Urinary IGFBP-3 immunoreactive profile was determined by neutral-size exclusion chromatography, followed by IGFBP-3 RIA analysis of the fractions. Urine showed a major peak of IGFBP-3 immunoreactivity around 17 kDa. The 17-kDa urinary IGFBP-3 chromatographic peak averaged 8461 ± 367 ng/12 h.m2 of body surface in healthy children, 3415 ± 739 in adults (P < 0.001), 2294 ± 354 in children with GHD before GH therapy (P < 0.001), and 7940 ± 1874 in children with GHD after GH therapy. Urinary IGFBP-3 was also measured by RIA in unfractionated urine; healthy children showed levels significantly higher (14575 ± 460 ng/12 h.m2) than adults (7823 ± 1083, P < 0.001) and higher than children with GHD before GH therapy (4710 ± 703, P < 0.001). Again, however, immunoreactive IGFBP-3 increased after GH treatment (12294 ± 3394). In the serum of the healthy children we characterized by specific IGFBP-3 WIB analysis, a 17.7-kDa immunoreactive form of IGFBP-3 that was absent in the serum of healthy adults and low in patients with GHD, increased during GH therapy. Serum samples were subjected to neutral-size exclusion chromatography and the fractions were analyzed by WIB.(ABSTRACT TRUNCATED AT 400 WORDS).",
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T2 - Relationship with GH therapy

AU - Spagnoli, Anna

AU - Gargosky, Sharron E.

AU - Spadoni, Gian Luigi

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AU - Oh, Youngman

AU - Boscherini, Brunetto

AU - Rosenfeld, Ron G.

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N2 - The insulin-like growth factor binding proteins (IGFBPs) are the carriers for insulin-like growth factor (IGF0-I and IGF-II. IGFBP-3 is GH-dependent and circulates associated with IGFs and an acid-labile subunit to form a 150-kilodalton (kDa) complex. In human serum, two immunoreactive molecular weight forms of IGFBP-3 have been identified. In human urine, radioimmunoassayable levels of IGFBP-3 have been detected. The objectives of this study were to characterize the molecular weight forms of IGFBP-3 in urine and serum of healthy children and adults and in children with GH deficiency (GHD), to quantify the urinary molecular weight forms of IGFBP-3, and to evaluate the relationship of these forms with GH therapy. Urine and serum were obtained from 12 prepubertal children with GHD, before and after 6 months of GH therapy, from 30 prepubertal healthy children, and from 8 healthy adults. Western immunoblotting (WIB) with IGFBP-3 antiserum (alpha IG-FBP-3g1) showed that in urine the most representative IGFBP-3 was a 17.7-kDa form. The 17.7-kDa IGFBP-3 was high in urine of healthy children compared with healthy adults and was low in children with GHD but increased after GH therapy. Urinary IGFBP-3 immunoreactive profile was determined by neutral-size exclusion chromatography, followed by IGFBP-3 RIA analysis of the fractions. Urine showed a major peak of IGFBP-3 immunoreactivity around 17 kDa. The 17-kDa urinary IGFBP-3 chromatographic peak averaged 8461 ± 367 ng/12 h.m2 of body surface in healthy children, 3415 ± 739 in adults (P < 0.001), 2294 ± 354 in children with GHD before GH therapy (P < 0.001), and 7940 ± 1874 in children with GHD after GH therapy. Urinary IGFBP-3 was also measured by RIA in unfractionated urine; healthy children showed levels significantly higher (14575 ± 460 ng/12 h.m2) than adults (7823 ± 1083, P < 0.001) and higher than children with GHD before GH therapy (4710 ± 703, P < 0.001). Again, however, immunoreactive IGFBP-3 increased after GH treatment (12294 ± 3394). In the serum of the healthy children we characterized by specific IGFBP-3 WIB analysis, a 17.7-kDa immunoreactive form of IGFBP-3 that was absent in the serum of healthy adults and low in patients with GHD, increased during GH therapy. Serum samples were subjected to neutral-size exclusion chromatography and the fractions were analyzed by WIB.(ABSTRACT TRUNCATED AT 400 WORDS).

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