A novel specific bioassay for serum human growth hormone
A novel specific bioassay for serum human growth hormone
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DOI：
10.1210/jc.85.11.4274
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发表时间：
2000-11-01
影响因子：
5.8
通讯作者：
Tanaka, T
中科院分区：
文献类型：
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作者：
Ishikawa, M;Nimura, A;Tanaka, T
人GH受体（hGHR）最近在Ba/F3细胞系上表达，Ba/F3细胞系是小鼠前B细胞淋巴瘤，其已被诱导成为克隆细胞系（Ba/F3-hGHR）。使用Ba/F3-hGHR细胞系，我们建立了血清hGH的生物测定法。hGH在1 ng至100 ng/mL浓度范围内以剂量依赖性方式刺激细胞增殖。除胰岛素样生长因子I（IGF-I）和GH结合蛋白外，生物测定中其他激素或生长因子均不影响细胞增殖。在该生物测定系统中，当浓度超过25.85 ng/mL时，游离IGF-I显著刺激Ba/F3-hGHR细胞增殖，但血清IGF-I不刺激细胞增殖，因为细胞增殖对血清中游离IGF-I的敏感性不足。然而，GH结合蛋白在最高浓度（100 ng/mL）下确实抑制细胞增殖，但在平均浓度（20 ng/mL）下不抑制细胞增殖。人血清刺激细胞增殖，这是完全抑制抗GH抗体。正常儿童和非GH缺乏性矮小症患者血清GH活性与免疫放射法（IRMA）测定的血清hGH浓度呈显著正相关（r = 0.967，r = 0.924，P < 0.0001）。正常儿童血清中的生物活性/IRMA比值为1.01 ± 0.26，非GH缺乏性身材矮小患者血清中GH刺激试验基础值和峰值分别为1.18 ± 0.24和1.00 ± 0.29。生物活性/IRMA比值为0.333 +/- 0.056（平均值+/- SD）。总之，我们建立了一种新的灵敏的hGH生物测定法，该方法对hGH体细胞生长作用具有特异性，可用于筛选生物活性hGH引起的身材矮小患者。
Human GH receptor (hGHR) was recently expressed on a Ba/F3 cell line, which is a mouse pro-B cell lymphoma that has been induced to become a cloned cell line (Ba/F3-hGHR). Using a Ba/F3-hGHR cell line, we have established a bioassay for serum hGH.hGH stimulated cell proliferation in a dose-dependent manner in concentrations ranging from 1 ng to 100 ng/mL. Cell proliferation was not influenced by other hormones or growth factors in the bioassay, with the exception of insulin-like growth factor I (IGF-I) and GH binding protein. Free IGF-I significantly stimulated the proliferation of Ba/F3-hGHR cells at concentrations over 25.85 ng/mL in this bioassay system, but serum IGF-I did not stimulate cell proliferation because the sensitivity of cell proliferation was insufficient for free IGF-I in serum. GH binding protein, however, did suppress cell proliferation at the highest concentration (100 ng/mL), but did not at the average concentration (20 ng/mL). Human serum stimulated cell proliferation, which was completely suppressed by anti-GH antibody. The GH bioactivity of serum samples from normal children and patients with non-GH deficient short stature correlated strongly with the serum hGH concentration determined by immunoradiometric assay (IRMA) (r = 0.967, r = 0.924, P < 0.0001, respectively). The ratio of bioactivity/IRMA was 1.01 +/- 0.26 in sera from normal children and 1.18 +/- 0.24 and 1.00 +/- 0.29 at basal values and peak values in GH stimulation tests, respectively, in sera from patients with non-GH deficient short stature. The bioactivity/IRMA ratio for the serum GH bioactivity of a patient who had biologically inactive GH caused by an amino acid substitution was 0.333 +/- 0.056 (mean +/- SD).In conclusion, we established a new sensitive bioassay for hGH that is specific for hGH somatogenic action and is useful for screening of patients with short stature caused by biologically inactive hGH.