Microassay for prostatic androgen receptors correlated with quantitative histological assessment.

A new microassay in which cryostat sections of prostate tissue were used to provide the source of soluble androgen receptor for biochemical assay, was devised using an isoelectric focusing method, with [3H]-mibolerone as the androgenic radioligand. Adjacent cryostat sections from the same tissue block were stained for diagnostic and quantitative histological assessment. The assay was used to illustrate variations in tissue androgen receptor concentration for correlation with epithelial cell content in benign prostate hyperplasia and prostatic cancer, and to show the effects of androgen receptor concentration of resection of prostatic tissue by electroresection. The results indicate that the heat in electroresection renders prostatic tissue unsuitable for androgen receptor assays, and suggest that knowledge of the cellular composition of carcinomatous prostates may be of importance in the full assessment of androgen receptor assay results. This method incorporates both a biochemical assay and histological assessment of the assayed tissue on near-facsimile sections, an advantage over conventional biochemical assays.

Immunohistology of oestrogen receptor and D5 antigen in breast cancer: correlation with oestrogen receptor content of adjacent cryostat sections assayed by radioligand binding and enzyme immunoassay.

Two monoclonal antibodies recognising epitopes associated with oestrogen receptor protein were evaluated against the assayable soluble oestrogen receptor concentration in a series of 149 breast carcinomas. One antibody (anti-ER) recognises the hormone binding unit of oestrogen receptor and gives nuclear staining; the other antibody (anti-D5) was raised to a component of soluble oestrogen receptor and gives cytoplasmic staining. To minimise variations attributable to tumour heterogeneity and sampling error immunohistology using the two monoclonal antibodies, radioligand binding assays, enzyme immunoassays, and quantitative histology were done on adjacent frozen sections. Thirty nine per cent, 48%, 54%, and 43% of the tumours were found to be oestrogen receptor positive by radioligand binding assay, anti-ER and anti-D5 immunohistology, and enzyme immunoassay, respectively. Strong correlations (p less than 0.0005) were found between anti-ER immunohistology and the radioligand binding assay. Only weak correlations were found between anti-D5 immunohistology and the results of other assay methods for oestrogen receptor. Nuclear staining of human breast cancers with the anti-ER monoclonal antibody thus seems to be an acceptable alternative to biochemical assays, with the additional advantage of showing intercellular and regional heterogeneity for oestrogen receptor content.

Clinicopathological significance of intratumoural variations in elastosis grades and the oestrogen receptor status of human breast carcinomas.

Cryostat sections from seventy-eight female breast carcinomas were assayed for oestrogen receptors by isoelectric focusing. Adjacent cryostat sections stained by Miller's elastic/van Gieson's method were graded for elastosis. Elastosis was similarly graded on near-equatorial paraffin sections from the same tumours. A positive correlation was obtained between elastosis in the near equatorial sections and oestrogen receptor positivity (p less than 0.0005), menstrual status (p less than 0.05) and parity (p less than 0.01) but no correlation was found between these factors and elastosis graded on cryostat sections from the more peripheral areas which had been selected for oestrogen receptor assay. These observations suggest that the central region of breast carcinomas, where connective tissue responses are fully developed, exhibits grades of elastosis with greater clinical significance. This may explain the conflicting published observations on the correlations between elastosis and oestrogen receptor status, which we believe are due to the lack of uniformity in tissue sampling. The possible implications of the absence of significant correlation between elastosis grades and tumour size, nodal status and disease-free interval are discussed.

Steroid hormone receptors in pituitary adenomas: a biochemical, immunohistochemical and morphometric study on cryostat sections.

Oestrogen receptors and progesterone receptors were measured by an isoelectric focussing technique in cytosols from cryostat sections of eight human pituitary adenomas. Cryostat sections adjacent to the assayed sample were stained for anterior pituitary hormones using the peroxidase-antiperoxidase technique, and the cellularity of each neoplasm was calculated with the aid of a computerized image analysis system. The results of ER and PR assays were adjusted to compensate for variations in cellularity. ER were present in three prolactin adenomas, one growth hormone adenoma and one gonadotrophin adenoma. The latter also contained PR. Steroid hormone receptors were not detected in two null cell adenomas and one non-secretory oncocytoma. The results support the suggestion that antihormonal chemotherapy may be valuable in the treatment of certain pituitary adenomas.

Cryostat section assay of oestrogen and progesterone receptors in meningiomas: a clinicopathological study.

Oestrogen receptors and progesterone receptors were measured in the cytosols from cryostat sections of 45 meningiomas from 40 patients (12 men, 28 women) using an isoelectric focusing technique. Near fascimile adjacent sections from the same tissue blocks were stained and examined to determine the histological subtype of the neoplasms. Appreciable levels of progesterone receptor (greater than 10 fmol/mg cytosol protein) were present in 24 (53.3%) of of the neoplasms, but no clinically important oestrogen receptor was detected in any of the tumours. Competitive binding studies on control tissue confirmed the specificity of the assay procedures. No correlation was found between progesterone receptor state and the age, sex, or menopausal state of the patients, or the histological subtype and site of the neoplasms. Four of the patients studied had multiple intracranial neoplasms, which in two were of differing progesterone receptor state. The presence of specific progesterone receptor in meningioma cytosols raises the possibility of hormonal manipulation in the treatment of this group of neoplasms.

Oestrogen receptor assay of cryostat sections of human breast carcinomas with simultaneous quantitative histology.

Cryostat sections of unfixed human breast carcinomas were assayed for oestrogen receptor (ER) content using an isoelectric focusing method to separate the receptor-bound oestradiol. Adjacent sections from the same tissue block were stained so that the tumour content could be estimated by point counting and the ER concentration adjusted to compensate for variations in cellularity. Elastosis was also assessed. The results confirm a positive correlation between ER values and both cellularity and elastosis. The measurement of ER in cryostat sections is relatively simple and rapid, is applicable to small tissue samples, and permits histological identification of the nature and composition of the assayed sample. The method is directly applicable to oestrogen receptor analyses of breast carcinomas in clinical laboratories with facilities for cryostat microtomy.

Immunohistochemical identification of adult and fetal haemopoiesis in the spleen in lymphoma, leukaemia and myeloproliferative disease.

Antisera to adult and fetal haemoglobins were used in the immunoperoxidase method to identify haemopoietic cells in spleens from patients with lymphoproliferative and myeloproliferative disorders. Splenic extramedullary haemopoiesis in adults often contains a major fetal component. Spleens infiltrated by Hodgkin's lymphoma are invariably negative, but other lymphomas, leukaemias and myeloproliferative diseases are commonly associated with splenic fetal haemopoiesis. Immunoperoxidase staining for haemoglobins is a useful way of determining the haemopoietic origin of round cell infiltrates in tissue sections.