Autoimmune anti-androgen-receptor antibodies in human serum.

Abstract

Circulating autoantibodies to human and rat androgen receptors are present at high titers in the blood sera of some patients with prostate diseases. The antibodies from some serum samples were associated with a purified IgG fraction and interacted with the 3.8S cytosolic androgen-receptor complexes of rat ventral prostate to form 9-to 12S units. Other serum samples, however, formed 14-to 19S units, suggesting that other immunoglobulins might be involved. In the presence of an anti-human immunoglobulin as a second antibody, the androgen-receptor-antibody complexes could be immunoprecipitated. The antibodies interacted with the nuclear and the cytosolic androgen-receptor complexes, either the DNA-binding or the nonbinding form, but not with receptors for estradiol, progestin, or dexamethasone from a variety of sources. Human testosterone/estradiol-binding globulin, rat epididymal androgen-binding protein, or rat prostate a-protein (a nonreceptor steroid-binding protein) also did not interact with the antibodies to form immunoprecipitates. About 37% of male and 3% of female serum samples screened had significant antibody titer. The chance ofrinding serum with a high titer is much better in males older than 66 years than in the younger males or females at all ages. The presence of the high-titer antibodies may make it possible to prepare monoclonal antibodies to androgen receptors without purification of the receptors for immunization.Steroid receptors are usually characterized by their ability to bind radioactive ligands. The isolation of polyclonal and monoclonal antibodies to estrogen (1, 2), glucocorticoid (3-5), and progestin (6-8) receptors in recent years, however, has provided alternative means to study these hormone receptors. An antibody against androgen receptors, however, has not been produced, mainly because of the difficulty in obtaining androgen receptors in sufficiently pure form and in amounts needed for efficient immunization. In this report, we show that anti-androgen receptor antibodies are present in high titers in blood sera of some male patients having clinical prostate problems. MATERIALS AND METHODSMaterials. 5a- [1,2,3,4,5,6,16,17-3H(N)

Effects of Estradiol and Testosterone on the Synthesis, Expression and Degradation of Androgen Receptor in Human Uterine Endometrial Fibroblasts

“…The cell lysate was spun at 800 g for 10 min to remove cell debris, and treated with 0.3% DCC at 4° C for 15 min to remove free steroids. The treated cell lysate (90 pi) was incu bated with 10 pi rat monoclonal AR antibody (Affinity Bio Reagents, Neshanic Station, N.J., USA) at a concentration of 4 pg/ml at 4°C for 16 h. The aliquot was incubated with 10 pi (= 10 pg) goat anti-rat IgG antibody (Organon Teknika Corp., Durham, N.C., USA) at 4°C for 16 h. The incubated aliquot was spun at 2,000 g for 10 min to precipitate the AR-AR antibody-rat IgG antibody complex [14], The effect of E and E + T on the degradation rate of AR was analyzed by the radioactivity of the precipitated AR complex.…”

Molecular Cloning of Human and Rat Complementary DNA Encoding Androgen Receptors

“…We had found earlier that some older men with prostate cancers had high titers of autoimmune antibodies to AR in their serum samples (20). Therefore, we investigated the possibility that the human autoantibodies could recognize the protein made by the reticulocyte lysate system.…”

“…In order to obtain a cDNA containing the amino acid-coding regions in different dones, we ligated two DNA fragments pre- (20 ,Lg) containing the 2.6-kb hAR DNA segment was linearized with restrction enzyme Bam HI, extracted with phenol-chloroform, and precipitated with ethanol. For transcription and translation, the procedures described by the supplier (Promega Biotec) of the polymerase and reticulocyte lysate were followed.…”

“…Antserum to human inmunoglobulin was used to precipitate human AR made by the translation of RNA transcribed from doned cDNA. The experiment was carried out as described elsewhere (20). Reticulocyte lysate containingtranslated AR was incubated with [3H]R1881 as shown in Tabkc 1.…”

Prediction of time to progression after orchiectomy by the nuclear androgen receptor content from multiple biopsy specimens in patients with advanced prostate cancer

“…For immunocytochemical assays monoclonal antibodies against the human androgen receptor have to be developed. As long as these do not become available, circulating antibodies against the human androgen receptor [20] may be of value.…”