Pituitary glycoprotein hormone alpha-subunit secretion by cirrhotic patients

Secretion of the alpha-subunit of pituitary glycoprotein hormones usually follows the secretion of intact gonadotropins and is increased in gonadal failure and decreased in isolated gonadotropin deficiency. The aim of the present study was to determine the levels of the alpha-subunit in the serum of patients with cirrhosis of the liver and to compare the results obtained for eugonadal cirrhotic patients with those obtained for cirrhotic patients with hypogonadotropic hypogonadism. Forty-seven of 63 patients with cirrhosis (74.6 percent) presented hypogonadism (which was central in 45 cases and primary in 2), 7 were eugonadal, and 9 women were in normal menopause. The serum alpha-subunit was measured by the fluorimetric method using monoclonal antibodies. Cross-reactivity with LH, TSH, FSH and hCG was 6.5, 1.2, 4.3 and 1.1 percent, respectively, with an intra-assay coefficient of variation (CV) of less than 5 percent and an interassay CV of 5 percent, and sensitivity limit of 4 ng/l. The serum alpha-subunit concentration ranged from 36 to 6253 ng/l, with a median of 273 ng/l. The median was 251 ng/l for patients with central hypogonadism and 198 ng/l for eugonadal patients. The correlation between the alpha-subunit and basal LH levels was significant both in the total sample (r = 0.48, P<0.01) and in the cirrhotic patients with central hypogonadism (r = 0.33, P = 0.02). Among men with central hypogonadism there was a negative correlation between alpha-subunit levels and total testosterone levels (r = 0.54, P<0.01) as well as free testosterone levels (r = -0.53, P<0.01). In conclusion, although the alpha-subunit levels are correlated with LH levels, at present they cannot be used as markers for hypogonadism in patients with cirrhosis of the liver

Monoclonal antibodies specific for the free alpha subunit of glycoprotein hormones and their use in the development of a sensitive immunofluorometric assay

Glicoprotein hormone free alpha subunit has been used as a marker for some pituitary tumors and to study the reactivity of glycoprotein hormone-producing cells under different circunstances. We describe a highly sensitive ans specific immunofluorometric assau for the measurement of serum free alpha subunit levels. The assay is based on a monoclonal antibody, specific for free alpha subunit, bound to microtiter plates. As tracer antibody we employed an europium-labelled free/complexed alpha subunit specific monoclonal antibody. Using overnight incubation and 50µl samples, the least detectable dose was of the order of 4 ng/1. Cross-reactivity with LH, TSH, FSH, and hCG was 6.5, 1.2, 4.3 and 1.1 percent, repectively. Normal adult males showed values ranging from 120 to 790ng/l, not different from normal adult premenopausal females (88 to 604 ng/l). In post-menopausal females, serum concentrations were significantly highler, ranging from 341 to 407 ng/l. In 56 patients with untreated pituitary tumors (18 "non-secreting", 25 GH-producing and 13 prolactin-producing tumors), 10 showed high values, 3 of them from the first group, 3 from the second and 4 from the third. We conclude that this highly sensitive assay can be a valualbe tool for the diagnosis and follow-up of selected patients with pituatary tumors and in other circumstances in which the glycoprotein hormone-producing cells of the pituitary require evaluation

Desenvolvimento de ensaio imunométrico para a dosagem sérica da subunidade alfa dos hormônios glicoproteicos / Development of an immunoflurometric assay for serum alfa subunit of the glycoprotein hormones

Tendo como base um anticorpo monoclonal e um policlonal, desenvolvemos um ensaio imunofluorométrico para a medida de subunidade alfa no soro. O anticorpo monoclonal contra subunidade alfa, empregado para captura, foi produzido a partir de um camundongo imunizado contr LH e apresenta reatividade cruzada contra as formas íntegras dos hormônios glicoproteicos. Já o anticorpo policlonal foi produzido em coelho imunizado com subunidade alfa livre e apresenta baixa reatividade cruzada com os hormônios íntegros. Para a revelaçäo da ligaçäo do policlonal à subunidade capturada pelo monoclonal, empregamos um monoclonal anti-IgG de coelho marcado com európio. O ensaio apresenta reatividade cruzada de 2,8 por cento com hCG, 3,2 por cento com FSH, 6,4 por cento com TSH e 9,4 por cento com LH. A sensibilidade é da ordem de 50 ng/L, com perfil de imprecisäo mostrando um erro intraensaio inferior a 10 por cento entre os valores 100 e 20.000 ng/L. O coeficiente de variaçäo interensaio mostrou-se inferior a 14 por cento dentro da faixa de valores encontrados. A definiçäo de valores normais foi feita a partir de uma populaçäo normal de 66 adultos, 33 homens e 33 mulheres na menacme, onde encontramos valores entre 70 e 902 ng/L, com mediana de 236 ng/L. Em 14 mulheres mnenopausadas econtramos valores entre 1.498 e 3.919ng/L. A aplicaçäo do ensaio em 33 pacientes com diagnóstico de tumor hipofisário (11 acromegálicos, 15 prolactinomas, 6 tumores "näo secretores" e 1 tumor produtor de TSH) mostrou valores elevados em 11 pacientes