Epithelial and muscle antigens in benign cystic teratomas of ovary.

Thirty beign cystic teratomas of the ovary were studied by immunofluroesecence, using specific human antisera for (a) squamous epithelial intercellular areas (pemphigus sera), (b) squamous epithelial basement membranes (bullous pemphigoid sera), (c) smooth muscle (chronic active hepatitis sera, and (d) striated muscle (thymona sera). Squamous epithelial intercellular staining was seen in all specimens and was present in the basal cell layers as well as in the prickle cell layers, suggesting a slow regeneration rate for the squamous epithelial lining of these tumors. Well-formed basement membranes were identified at the periphery of all squamous epithelial structures. Smooth muscle was found in 73% of the neoplasms, and the immunoflurorescent findings correlated with light microscopy. No skeletal muscle components were identified. Within the limits of the antibodies used and the antigens detected, these resutlts support the concept that benign ovarian cystic teratomas are composed of mature, heterotopic tissue elements.

Pulmonary hemorrhage in systemic lupus erythematosus.

Massive pulmonary hemorrhage confirmed by pathologic examination occurred in three of 12 patients with systemic lupus erythematosus (SLE) complicated by lung disease. Two patients died. Hemorrhage was clinically unrecognized in two cases. The third patient survived after an open lung biopsy and an increase in prednisone therapy. In the absence of hemoptysis, awareness of the possibility of pulmonary hemorrhage in patients with SLE and a high index of suspicion are necessary for early recognition of this uncommon but potentially fatal complication.

Immunofluorescent detection of angiotensin-converting enzyme (ACE) in Gaucher cells.

The cellular localization of the elevated angiotensin-converting enzyme (ACE) in the spleen of a patient with Gaucher's disease was examined by immunofluorescence using an antibody prepared against highly purified human lung enzyme. Intense ACE-specific cytoplasmic fluorescence was observed in Gaucher cells, but not in various controls, indicating the localization of large quantities of enzyme in these cells. These results and the demonstrated capability of mononuclear phagocytes for marked induction of this enzyme suggest the possibility that induction of active synthesis of ACE in Gaucher cells may be responsible for the elevated enzyme levels in the serum and spleen of patients with Gaucher's disease.

Quantitation of the immunocytochemical assay for estrogen receptor protein (ER-ICA) in human breast cancer by television imaging.

A Quantimet 720D Image Analysis System has been programmed for light microscopic evaluation of the nuclear estrogen receptor distribution in frozen sections of human breast cancer stained by the peroxidase-antiperoxidase method using monoclonal antibodies to estrogen receptor protein (ER). This method provides precise criteria for distinguishing ER-positive and -negative cells and a sensitive and reproducible means for densitometric quantification of the staining patterns. Although imaging sequence and graphic analysis are automated by computer programs, light pen interaction provides supervision of feature selection. Imaging of the immunocytochemical assay (ER-ICA) in 50 patients revealed marked heterogeneity of nuclear estrogen receptor concentration varying over a nearly 100 fold concentration range. Various ER concentration patterns were evident: (I) distributions with a single peak (CV = 5%) present at various concentration levels; (II) bimodal distributions, revealing co-existent ER-positive and ER-negative subpopulations; (III) multimodal distributions with a number of resolvable concentration peaks; and (IV) highly skewed distributions with or without discernible peaks, frequently extending over the entire concentration range. Statistical methods of de-convolution were applied to determine the frequency and ER concentration characteristics of component subpopulations in the mosaic cases and for resolving the proportion of ER-positive and -negative cells. An approach for evaluating nuclear ER content in conjunction with ER concentration patterns in individual patients revealed whether spread in the ER concentration distribution resulted from differences in nuclear ER content or from variability in nuclear volume distribution.

Histochemical analyses of steroid hormone receptors in breast and prostatic carcinoma.

Histochemical analyses estrogen (ER) and progesterone (PgR) receptors in breast cancer were statistically correlated with results of dextran-coated charcoal (DDC) and sucrose gradient assays. Correlated for ER was 91% of 363 cases, and for PgR 88% of 255 specimens. Breast cancer ER/PgR positivity by histochemistry correlated with a favorable clinical response to endocrine therapies in 72% of 25 cases, while ER/PgR negativity correlated with a lack of response in 96% of 22 cases with Stage IV disease. Nuclear ER/PgR correlated with a poor response to therapy in 8 of 12 patients. An in vitro technique to detect nuclear translocation of ER revealed two groups of ER positive cases, with 11 of 17 exhibiting translocation and 6 not displaying translocation. In prostatic carcinoma, 72% of 65 men were positive for ER and/or androgen receptor. Comparison of specimens obtained without and with electrocautery revealed a preponderance of nuclear binding in the latter, suggesting heat-induced nuclear translocation of receptor. coumestrol, a naturally fluorescent, entirely unaltered estrogen was also used for histochemical detection of ER. Results correlated with ER by DCC in 87% of 61 breast cancers. Coumestrol was additionally used to visually observe receptor and nuclear translocation of ER in intact whole cells in culture.

Immunofluorescence of soft-tissue tumors with anti-smooth-muscle and anti-skeletal-muscle antibodies.

One hundred thirteen soft-tissue tumors were studied by immunofluorescence using anti-smooth-muscle (SMA) and anti-skeletal-muscle antibodies (MGA). Of the classic leiomyomas examined, 94% manifested bright fluorescence with SMA, whereas cellular leiomyomas and leiomyosarcomas failed to stain. Of the rhabdomyosarcomas studied, 21% showed fluorescent cross-striations or fluorescent granules after incubation with MGA. None of 48 control tumors fluoresced with either antibody. No tumor vessel seen showed smooth-muscle fluorescence in its walls. Although of little diagnostic value, immunofluorescence utilizing specific human antibodies is a valuable tool with which to study some of the antigenic components of these neoplasms.

Immunohistologic diagnosis of sarcoidosis. Detection of angiotensin-converting enzyme in sarcoid granulomas.

Angiotensin-converting enzyme (ACE) was detected by indirect immunofluorescence examination. ACE was present in the cytoplasm of epithelioid cells in granulomas from 38 of 39 patients who had clinical sarcoidosis but not in any of 37 nonsarcoid granulomas, normal lymph nodes, or splenic controls. Intensity of staining with ACE antiserum was inversely proportional to the degree of fibrosis and was significantly less intense in fibrotic granulomas. The detection of ACE appears to be a distinct aid in establishing a definitive tissue diagnosis of sarcoidosis.

An improved histochemical method for detection of estrogen receptors in mammary cancer. Comparison with biochemical assay.

Unselected, consecutive surgical specimens from 120 women with cancer of the breast were subjected to histochemical assay for the presence of estrogen receptor. A fluoresceinated bovine serum albumin--estradiol conjugate was used that linked estradiol at position 17 and contained 5 mol fluorescein and 4 mol estradiol per mole albumin. Simultaneous competitive binding studies with excess unlabeled estradiol, diethylstilbestrol, and the antiestrogen nitromifene citrate were regularly performed. Results were compared to those obtained by the dextran-coated charcoal receptor assay. Three specimens were necrotic, two others thawed, and two lacked sufficient protein for biochemical analysis. One specimen did not contain tumor, and 11 others showed a predominant nuclear staining pattern. Nuclear receptor was not assayed biochemically. Comparison of results in the remaining 101 cases showed agreement in 92%. The precedure is uncomplicated, economical, and could be performed and interpreted in any pathology laboratory.

Glomerulonephritis due to Staphylococcus aureus antigen.

Staphylococcus aureus antigen was identified within the glomeruli of a patient with acute bacterial endocarditis and diffuse glomerulonephritis. Routine immunofluorescence had revealed only granular deposits of complement (C3). C3 activator and C4 were not present. Direct immunofluorescence studies with a specific anti-Staphylococcus aureus conjugate were positive. Electron microscopy showed subepithelial and intramembranous electron-dense deposits. Eluates of the kidney did not contain anti-S. aureus antibody. The absence of immunoglobin suggests that the toxic action of S. aureus antigens may activate complement and cause glomerular injury and that immune complexes are not essential for the production of glomerulonephritis in this entity.

Acute pulmonary complications in systemic lupus erythematosus. Immunofluorescence and light microscopic study.

Lung tissue obtained from eight consecutive patients with systemic lupus erythematosus complicated by severe, acute pulmonary disease was studied by both light and immunofluorescence microscopy. Light microscopic examination disclosed interstitial pneumonia in four cases, cytomegalovirus pneumonitis in one case, bronchiolitis and peribronchiolitis in one case, pulmonary infarction in one case and focal atelectasis in the remaining case. Direct immunofluorescence examination revealed focally bound immunoglobulins or complement (C3) within pleural and/or pneumocyte nuclei in each specimen. Immunohistologic studies in these cases may thus suggest a diagnosis of systemic lupus erythematosus with acute pulmonary complications, despite the lack of specificity of the pathologic changes seen by light microscopy.