Monoclonal antibodies directed against preneoplastic and neoplastic murine mammary lesions.

We have produced a panel of monoclonal antibodies directed against a dimethylbenzanthracene-induced murine mammary tumor. Five rat-mouse hybridomas produced antibodies that bound to some murine mammary tumors, but not to normal renal adherent cells, lymphocytes, 3T3 fibroblasts, red blood cells, or mammary gland. One of these antibodies, designated AMT8, was selected for further evaluation based on its relatively strong reactivity, as determined by immunofluorescence. Indirect immunofluorescent studies on frozen histological tissue sections and quantitative immunofluorescent binding studies on cultured normal and tumor cells revealed that AMT8 was bound to certain murine mammary tumors and their preneoplastic hyperplastic nodules, but not to normal murine organs including normal mammary glands. Two tumors and their hyperplastic alveolar nodule counterparts that contained the antigen recognized by AMT8 did not express functional estrogen and progesterone receptors, indicating that antigen expression was not dependent on functional receptors. The antigen recognized did not cap, was found to modulate slowly, and was reexpressed in the presence of excess AMT8. From these findings, we conclude that AMT8 may prove to be a valuable tool for the study of early mammary tumorigenesis.

Tumor-associated antigen expression of primary and metastatic colon carcinomas detected by monoclonal antibody 17-1A.

Murine monoclonal antibody (MAb) 17-1A has been used in radioimmunodetection and immunotherapy trials of intestinal adenocarcinoma in humans. Tumor heterogeneity of antigen expression has been recognized as a potential limiting factor in such studies. The authors report a study designed to evaluate the degree of heterogeneity of 17-1A antigen expression among primary and metastatic human colon carcinomas. All 141 specimens, including 74 primary or metastatic colonic adenocarcinomas, were evaluated with the use of an avidin-biotin complex immunoperoxidase technic on briefly fixed frozen tissue sections. All of these showed at least focal staining with MAb 17-1A. However, well- or moderately differentiated tumors generally showed diffuse cytoplasmic immunostaining, whereas poorly differentiated tumors showed minimal immunostaining with no detectable antigen in most areas. In 16 cases that had both primary and metastatic adenocarcinomas or multiple metastatic tumors, 17-1A antigen expression was similar among the tumor sites except for one case. This case showed variation in tumor differentiation and corresponding variation in 17-1A antigen expression. Of 36 additional malignant tumors that were not of colonic epithelial origin, adenocarcinomas of the stomach, duodenum, endometrium, ovary, and breast showed 17-1A antigen expression.

Intraoperative probe-directed immunodetection using a monoclonal antibody.

To assess monoclonal antibody (MAb) 17-1A and its F(ab')2 fragment in intraoperative radioimmunodetection and to evaluate further the clinical usefulness of a hand-held gamma-detecting probe (GDP), we injected radiolabeled monoclonal antibody 17-1A three to six days preoperatively or its F(ab')2 fragment two to three days preoperatively into 18 patients with colorectal cancer. Intraoperative GDP counts with tumor-tissue ratios of 1.5:1 or greater were obtained from 15 (75%) of 20 tumor sites, with ratios averaging 2.3:1 for fragments and 3.4:1 for whole antibody. The GDP counts contributed to intraoperative decision making in three patients, either by localization of tumor not identified by inspection or palpation or by mapping margins of resection with histologic confirmation of a local/regional recurrence. These preliminary data demonstrate that probe-directed, intraoperative radioimmunodetection can assist the surgeon in detecting subclinical tumor deposits and thus better evaluate the extent of primary or recurrent colorectal cancers intraoperatively.

Radioimmunoguided surgery using monoclonal antibody.

The potential proficiency of radioimmunoguided surgery in the intraoperative detection of tumors was assessed using labeled monoclonal antibody B72.3 in 66 patients with tissue-proved tumor. Monoclonal antibody B72.3 was injected 5 to 42 days preoperatively, and the hand-held gamma-detecting probe was used intraoperatively to detect the presence of tumor. Intraoperative probe counts of less than 20 every 2 seconds, or tumor-to-adjacent normal tissue ratios less than 2:1 were considered negative (system failure). Positive probe counts were detected in 5 of 6 patients with primary colon cancer (83 percent), in 31 of 39 patients with recurrent colon cancer (79 percent), in 4 of 5 patients with gastric cancer (80 percent), in 3 of 8 patients with breast cancer (37.5 percent), and in 4 of 8 patients with ovarian cancer (50 percent) undergoing second-look procedures. Additional patients in each group were scored as borderline positive. Overall, radioimmunoguided surgery using B72.3 identified tumors in 47 patients (71.2 percent), bordered on positive in 6 patients (9.1 percent), and failed to identify tumor in 13 patients (19.7 percent). Improved selection of patients for antigen-positive tumors, the use of higher affinity second-generation antibodies, alternate routes of antibody administration, alternate radionuclides, and more sophisticatedly bioengineered antibodies and antibody combinations should all lead to improvements in radioimmunoguided surgery.

A unique familial lobular glomerulopathy.

Five members of a family (four brothers and their father) were found to have proteinuria and microhematuria. One of the affected brothers developed chronic renal failure. Although a renal biopsy was never performed, this brother has subsequently received a renal transplant from a clinically unaffected sister. Renal biopsy was performed on four of the siblings (three brothers and the sister). Renal tissue from all three brothers who underwent biopsy showed similar glomerular lesions, characterized by striking cloverleaf expansion of glomerular lobules by an acellular hyaline material and segmental mesangial cell proliferation. Electron microscopy showed massive mesangial and subendothelial deposits and occasional areas of glomerular basement membrane splitting with round inclusions. The renal biopsy specimen of the sister was normal on light and electron microscopy and negative for immunoglobulins and complement on immunofluorescent staining. Immunofluorescence microscopy on one of the male siblings was negative for immunoglobulins and complement. No other physical or serologic abnormalities could be established. To our knowledge, this is a unique form of familial glomerulopathy that has not been previously described.

Heterologous carcinosarcoma of the fallopian tube. A case report.

Carcinosarcomas occur infrequently in the female genital tract. They arise predominantly in the uterus and much less frequently in the vagina, cervix and ovary. They rarely occur in the fallopian tube; only 25 such cases have been reported in the literature to date. The rarity of carcinosarcoma of the fallopian tube prompted this case report.

Granular-cell tumor of the breast. A cytologic, immunohistochemical and ultrastructural study of two cases.

Two primary granular-cell tumors of the breast are reported. Cytologically and histologically, these tumors had the classic features of small, eccentrically located nuclei and numerous periodic acid-Schiff-positive cytoplasmic granules. The characteristic cytologic features were best appreciated in touch imprints, not in frozen sections. Immunohistochemically, the tumors demonstrated diffuse cytoplasmic staining for S-100 protein but negative staining for myoglobin. The significance of these immunohistochemical staining characteristics, particularly in evaluating the possible histogenesis of these tumors, is discussed. The ultrastructural features of these two tumors are also presented and compared to findings reported by other investigators.

The response of a non-functional VIP- and somatostatin-containing tumour to tolbutamide in vitro.

A patient with a tumour containing clinically non-expressive somatostatin (SRIF) and vasoactive intestinal peptide (VIP) was studied in vivo with basal and tolbutamide-provoked SRIF and VIP measurements and failed to respond to tolbutamide infusion. An acute cell dispersion model was used to study this tumour after resection. Incubation of tumour cells in tolbutamide (2 mg/ml) resulted in increases in intracellular SRIF but not in the levels of SRIF released into the incubating medium. In contrast, incubation of tumour cells with tolbutamide decreased supernatant (extracellular) and total (intracellular) VIP by 50%, suggesting a local peptide-peptide modulation of VIP release by high intracellular levels of SRIF or, alternatively, suppression of VIP synthesis and/or release by tolbutamide. Failure of 'nonfunctional' tumours to produce symptoms or abnormal plasma peptide levels may be due to defects in peptide release or complex paracrine peptide-peptide interactions.

Diagnostic histochemical and immunohistochemical studies in malignant mesothelioma.

This study was undertaken to evaluate the relative utility of histochemical and immunohistochemical stains in diagnosing malignant mesothelioma of the thorax. We performed a battery of histochemical stains, including periodic acid Schiff (PAS) with and without diastase, mucicarmine, colloidal iron (Coll Fe) with and without hyaluronidase, and immunohistochemical stains for keratin and carcinoembryonic antigen (CEA) on 12 pleural mesothelioma specimens obtained from 11 patients, five primary pulmonary adenocarcinomas, and one metastatic adenocarcinoma each to pleura and pericardium. All diagnoses were established by autopsy or thorough clinical and surgical evaluation. The diagnosis of mesothelioma was established following rigid anatomic criteria. All tissue was formalin fixed and paraffin embedded. Commercially available reagents and antisera were used in all cases. Results showed a high rate of positivity for keratin and hyaluronidase-sensitive Coll Fe in the mesotheliomas while adenocarcinomas were uniformly positive for CEA and keratin and generally positive for PAS-D (diastase) and mucicarmine. Mesotheliomas were negative for CEA, mucin, and PAS-D. Positive keratin staining was also seen in the spindle cell components of mesotheliomas. Immunohistochemical stains often added significantly to our ability to establish the diagnosis of mesothelioma with confidence, since they were more frequently and more clearly positive than histochemical stains.

Prostatic carcinoma metastatic to bone: sensitivity and specificity of prostate-specific antigen and prostatic acid phosphatase in decalcified material.

Decalcified bone marrow biopsies containing metastatic tumor from 36 patients were stained for prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) using the avidin biotin complex (ABC) immunoperoxidase technique. Of these patients, 22 had known prostate primaries, ten had known nonprostatic, and four female patients had unknown primaries. Prostate-specific antigen was identified in 86% (19/22) of the metastatic prostatic carcinomas. Prostatic acid phosphatase was present in only 36% (8/22). None of the patients with nonprostatic primaries or unknown primaries showed positive staining for either antigen (0/14). This study indicates that immunoperoxidase staining for PSA is very sensitive and specific in the diagnosis of metastatic prostate carcinoma, while PAP was less sensitive using decalcified bone marrow specimens. We believe that immunostaining with PSA should be of great value in diagnosis of prostatic carcinoma metastatic to the bone.

Distinguishing malignant mesothelioma from pulmonary adenocarcinoma: an immuno-histochemical approach using a panel of monoclonal antibodies.

A panel of six monoclonal antibodies (MAbs) was employed to evaluate antigen expression in pulmonary adenocarcinomas and mesotheliomas. Monoclonal anti-human milk fat globulin (HMFG-2), anti-carcinoembryonic antigen (NP-2), anti-epithelial membrane antigen (EMA), anti-cytokeratin (PKK-1), anti-tumor-associated antigen 72 (B72.3), and anti-human myelomonocytic antigen (Leu M-1) antibodies were used to localize their respective antigens in formalin-fixed, paraffin-embedded tumors by using the avidin-biotin-complex immunoperoxidase technique. In all, 28 mesotheliomas obtained from Ohio State University Anatomic Pathology files and from a Southwest Oncology Group (SWOG) protocol were compared to 22 pulmonary adenocarcinomas by using this MAb panel. None of the mesotheliomas demonstrated positive staining with MAbs NP-2 (anti-CEA) or Leu M-1. However, 95% (21/22) of adenocarcinomas stained with one of these two antibodies. Although neither of these two MAbs stained all adenocarcinomas, each antibody demonstrated positive immunostaining in more than 90% of the adenocarcinomas studied. Therefore, MABs NP-2 and Leu M-1 are, individually, quite useful for distinguishing mesothelioma from adenocarcinoma. However, in our study, no single MAb could be used to distinguish these two tumor types in every case. MAb B72.3 stained 91% (20/21) adenocarcinomas but also stained 7% (2/28) of mesotheliomas. MAb HMFG-2 reacted positively with 95% of adenocarcinomas, but also stained 39% of the mesotheliomas, usually in a membranous pattern. MAbs EMA and PKK-1 were not found useful in distinguishing mesothelioma from adenocarcinoma. We conclude that MAbs Leu M-1 and NP-2 were both useful in distinguishing mesothelioma from pulmonary adenocarcinoma in that positive staining was demonstrated in adenocarcinomas and not mesotheliomas.

Acute amino acid nephrotoxicity.

Previous clinical and experimental data suggest that a limited number of AAs with particular molecular characteristics can adversely affect renal function. The present investigation was conducted to better define the scope and the molecular basis of this nephrotoxicity. Sprague-Dawley rats (N = 31) undergoing a solute diuresis were subjected to 80 min infusions (125 mumol/kg/min) of cationic AAs (lysine, arginine), anionic AAs (glutamic acid, aspartic acid), or neutral AAs (alanine, glycine). Rats infused for 80 min with equimolar amounts of urea or dextrose served as controls (N = 8). GFR (Cioth) were measured 40 min before, during, and 140 min after these infusions. Albumin excretion rates were determined by radioimmunoassay. All AA infusates induced significant (p less than 0.001) reductions in GFR compared to the control infusates (cationic AAs: 62% +/- 4; anionic AAs: 57% +/- 5; neutral AAs: 33% +/- 1; controls: 8% +/- 4) (X +/- S.E.M.). However, only cationic AAs induced a significant increase in albumin excretion (360% +/- 72; p less than 0.001). AA-treated rats had histologic changes with mild tubular injury compared to control rats. These data indicate the following. (1) AAs have in common a nephrotoxic potential. (2) This nephrotoxicity arises, in part, from the nonvariable portion of AA molecules since glycine, which has no variable (R) group, induced a significant reduction in GFR (32% +/- 1). (3) The ability of AAs to decrease GFR is enhanced by certain R groups. However, R group charge is not a critical factor in producing this response. (4) AA-induced increments in albumin excretion and reductions in GFR must arise via independent mechanisms. Since AA infusions are commonly used in patients with ARF the possibility that such therapy might have a deleterious effect on renal function and on the subsequent recovery from ARF should be entertained.

Glomerular basement membrane splitting and microaneurysm formation associated with nitrosourea therapy.

A patient who developed renal insufficiency following nitrosourea therapy is reported. Light, immunohistochemical, and electron microscopic studies of the renal biopsy disclosed an unusual glomerular basement membrane injury. Light microscopy showed extensive basement membrane splitting and capillary aneurysm formation. Electron microscopic examination revealed an extensive subendothelial accumulation of electron-lucent granular material. The glomerular basement membrane was separated from the mesangium and showed splitting of the lamina densa. Immunofluorescent and immunoperoxidase staining of the glomeruli was negative for immunoglobulin, complement, and fibrinogen. This form of nitrosourea-associated glomerular injury has not been described previously.

Mitomycin-C induced improvement in superficial bladder tumors: a morphologic and immunohistochemical evaluation.

Twenty bladder biopsies from ten patients with superficial transitional cell carcinoma were obtained prior to and following therapy with intravesical mitomycin-C. Each biopsy was evaluated histologically and immunohistochemically for A, B, H blood group antigen (BGA) expression. A, B and H blood group antigens were identified using monoclonal anti-A, B and H antibody and were localized with the avidin-biotin-peroxidase complex (ABC) technique. Particular attention was directed toward flat urothelial lesions and the urothelium adjacent to papillary tumors. Routine histologic evaluation showed improvement in post-therapy biopsies compared to corresponding pre-therapy biopsies in 7/10 cases but was equivocal in 3/10 cases. Immunohistochemical evaluation, however, showed improvement in all 10 cases, as judged by increased urothelial BGA expression. This increase in urothelial BGA expression after intravesical mitomycin-C therapy suggests a therapy induced improvement in dysplastic urothelium which was not uniformly evident on routine histologic examination.

Immunohistochemical evaluation of human placental implantation: an initial study.

Immunohistochemical staining for human chorionic gonadotropin and factor VIII-related antigen with the avidin-biotin complex immunoperoxidase technique was used as a marker for syncytiotrophoblast and endothelial cells, respectively, in the human placental bed. Material from placental implantation sites at varying stages of gestation (8 weeks to term) was studied. Trophoblastic invasion of the uterine stroma and blood vessels were evaluated. Syncytiotrophoblasts lining placental villi and anchoring villi were positive for human chorionic gonadotropin at all stages of gestation studied. Endothelial cells lining maternal uterine blood vessels were positive for factor VIII-related antigen. At early stages of intrauterine placentation (8 and 11 weeks) trophoblastic invasion of uterine blood vessels and trophoblastic incorporation in the walls of dilated vessels were present. An unexpected finding, however, was the large number of giant cells in the superficial placental bed which had morphology suggestive of syncytiotrophoblast but which were negative for human chorionic gonadotropin. In addition, many enlarged, rather pleomorphic cells lining superficial blood vessels were found to be positive for factor VIII-related antigen, which identified them as endothelial cells and not migrating trophoblastic elements. This study demonstrates that human chorionic gonadotropin and factor VIII-related antigen immunoperoxidase staining is a helpful adjunct in evaluating human placentation and suggests extension of the technique with use of other antibodies to evaluate components of the placental bed.

ABO (H) cell surface antigens in benign and malignant parotid neoplasms.

Twenty-two inflammatory, benign, or malignant parotid lesions were studied by means of the specific red cell adherence test (SRCA), a modification of the Coombs' mixed cell agglutination reaction. In 22 normal parotid tissues the acinar structures were devoid of red cell agglutination, but it was present in ductal epithelium. Findings were similar in 2 cases of parotitis, 11 benign mixed tumors, and 1 malignant mixed tumor. All lacked red cell agglutination in areas of neoplastic change. Benign Warthin's tumors (4 cases) demonstrated antigenicity in the columnar epithelial component of the tumor, but lacked red cell agglutination in areas of the lymphoid component. One malignant Warthin's tumor showed agglutination in areas of normal columnar epithelium but not in areas of malignant dedifferentiation. Undifferentiated carcinoma (1 case) and adenoid cystic carcinoma (2 cases) did not possess detectable ABO (H) antigens in neoplastic areas of the gland. The absence of ABO antigens in normal acinar glands supports their suggested myoepithelial or mesenchymal derivation, as the absence of antigen in benign and malignant mixed tumors supports their proposed mesenchymal derivation. Ductular epithelium and the epithelial components of benign Warthin's tumors have ABO (H) antigens, while the loss of antigen in the epithelial portion of the malignant Warthin's tumor is characteristic of epithelial neoplastic dedifferentiation. Loss of antigen in adenoid cystic and undifferentiated carcinomas of the parotid supports the concept that antigen is absent in epithelially derived malignant neoplasms.

Alveolar soft part sarcoma of the vagina: an immunohistochemical and electron microscopic study.

Alveolar soft part sarcoma is a soft tissue neoplasm of unknown histogenesis which has a distinctive morphology. It is a relatively rare tumor with approximately 200 cases described in the literature. Only two cases have previously been reported as occurring in the vagina. The purpose of this paper is to report the third case of alveolar soft part sarcoma occurring in the vagina and to emphasize the role of electron microscopy in the differential diagnosis.

Extra-adrenal paraganglioma and pulmonary chondroma: a case report and review of the literature.

This report describes the findings of an extra-adrenal paraganglioma and chondroma of the lung in an elderly man. Tissue analysis of the paraganglioma showed a high level of catecholamines, as well as the presence of somatostatin and small amounts of other peptide hormones. Immunoperoxidase staining demonstrated the distribution of peptide hormones in the tumor. This case illustrates two important points. First, it confirms the ability of paraganglionic tissue to secrete peptide hormones. Secondary, it adds yet another case to the unusual group of patients described by Carney (Carney JA: Cancer 43:374-382, 1979).