Tumor-associated glycoprotein (TAG-72) in ovarian carcinomas defined by monoclonal antibody B72.3.

Murine IgG1 monoclonal antibody (MoAb) B72.3, reactive with a high molecular weight glycoprotein complex [designated tumor-associated glycoprotein-72 (TAG-72)] was shown, with the use of the avidin-biotin-complex-immunoperoxidase technique and surgically resected tissues, to be reactive with a variety of histologic tumor types. TAG-72 is expressed in at least 5% (and up to 100%) of the malignant epithelial cells in 77% (n = 52) of human primary cancers and 71% (n = 31) of metastatic ovarian cancers of the common "epithelia" histologic category. Of these, several histologic types, including serous and mucinous cystadenocarcinomas, undifferentiated carcinomas, and less common types of ovarian carcinoma, were all shown to express TAG-72. In contrast, normal ovarian tissues and 26 of 27 benign ovarian tumors of various histologic types failed to express similar levels of TAG-72. Of interest is the 1 benign tumor that demonstrated unusual glandular complexity, as well as 3 tumors designated as borderline malignancy, that contained elevated TAG-72 expression. MoAb B72.3 also was used successfully to detect ovarian carcinoma cells in 28 cytologic preparations of human serous effusions and peritoneal washings. The reactivity of MoAb B72.3 was shown to be distinct from that of MoAb OC125 and an anti-CEA MoAb B1.1. The potential applications of MoAb B72.3 in the study of human ovarian cancer cell populations, as well as in several aspects of the management of human ovarian cancer, are discussed.

Selective interferon-induced enhancement of tumor-associated antigens on a spectrum of freshly isolated human adenocarcinoma cells.

Freshly isolated cells from patients with pleural or peritoneal effusions cytologically diagnosed as adenocarcinoma (n = 43), malignant nonepithelial neoplasms (n = 10), and benign (n = 8) were analyzed for expression of constitutive levels of the tumor antigens TAG-72 [recognized by monoclonal antibody (MAb) B72.3] and carcinoembryonic antigen (CEA) (recognized by MAb COL-4) as well as the class I and class II major histocompatibility (MHC) antigens, and the ability of human interferons (Hu-IFNs) to enhance cell surface expression of those antigens as measured by MAb binding. Both type I and type II IFNs enhanced the expression of TAG-72 and CEA and altered the level of expression of the MHC antigens. Comparative studies of three different Hu-IFNs (IFN-alpha A, IFN-beta ser, and IFN-gamma) revealed that IFN-gamma was the most potent in augmenting either B72.3 or COL-4 binding. Unlike the IFN-gamma -mediated induction of the class II human leukocyte antigens, the change in tumor antigen expression consisted of enhanced constitutive antigen expression; de novo induction of either TAG-72 or CEA could not be achieved by either type I or type II IFN. Of 43 effusions isolated from different adenocarcinoma patients, 42 (97.7%) expressed either CEA or TAG-72, and treatment with Hu-IFN increased the level of expression of either antigen in 36 of 42 samples (85.7%). These studies demonstrate the augmentation of tumor-associated antigens on human carcinoma cells isolated from serous effusions by Hu-IFNs which may be used to enhance the targeting of conjugated MAbs to human carcinoma lesions.

Distribution of oncofetal antigen tumor-associated glycoprotein-72 defined by monoclonal antibody B72.3.

Murine monoclonal antibody B72.3, prepared against a membrane-enriched extract of human metastatic carcinoma, was reacted with a spectrum of adult and fetal human tissues using avidin-biotin-complex immunohistochemical techniques to evaluate the expression of the reactive tumor associated glycoprotein (TAG)-72 antigen. TAG-72 was shown to be expressed in several epithelial-derived cancers including 94% of colonic adenocarcinomas, 84% of invasive ductal carcinomas of the breast, 96% of non-small cell lung carcinomas, 100% of common epithelial ovarian carcinomas, as well as the majority of pancreatic, gastric, and esophageal cancers evaluated. TAG-72 expression was not observed, however, in tumors of neural, hematopoietic, or sarcomatous derivation, suggesting that the TAG-72 antigen is "pancarcinoma" in nature. Appreciable monoclonal antibody B72.3 reactivity was generally not observed in adult normal tissues, with limited reactivity noted in a few benign lesions of the breast and colon. TAG-72 antigen expression was detected, however, in fetal colon, stomach, and esophagus, thus defining TAG-72 as an oncofetal antigen. TAG-72 has previously been shown to be distinct from carcinoembryonic antigen and other tumor associated antigens. The pancarcinoma distribution and lack of significant reactivity with normal adult tissues of monoclonal antibody B72.3 suggest its potential diagnostic and therapeutic utility for human carcinomas.

Phenotypic characterization of lung cancers in fine needle aspiration biopsies using monoclonal antibody B72.3.

Monoclonal antibody B72.3, reactive with a high-molecular-weight glycoprotein tumor-associated antigen (designated TAG-72), has been previously shown to be reactive with formalin-fixed paraffin-embedded tissue sections of adenocarcinomas of the ovary, colon, and breast and not a variety of normal adult tissues. It has demonstrated utility as an immunocytochemical adjunct to diagnose carcinoma in cell block and cytocentrifuge preparations of human serous effusions, with selective reactivity for tumor cells (particularly adenocarcinoma) over reactive mesothelium. In this study, fine needle aspiration biopsies of 127 lung cancers (93 primary and 34 metastatic tumors) as well as 18 benign lung lesions were analyzed with monoclonal antibody B72.3 using avidin-biotin-peroxidase techniques. Monoclonal antibody B72.3 showed reactivity with 100% of the 27 lung adenocarcinomas and adenosquamous carcinomas, with greater than or equal to 10% of tumor cells showing reactivity in 22 of 27. A lesser percentage of squamous cell carcinomas (24 of 31) and large cell carcinomas (7 of 13) were immunoreactive, and of these several were weakly reactive with less than or equal to 1% tumor cells reacting. In contrast, monoclonal antibody B72.3 failed to react with any of the 21 small cell carcinomas or one carcinoid tumor evaluated. In 35 patients tumor-bearing tissue was resected, and formalin-fixed tissue sections were also evaluated. The staining pattern and percentage of tumor cells positive in the aspiration biopsies were, in most cases, highly predictive of the reactivity observed in corresponding resected tumor. Metastatic adenocarcinomas to lung from various body sites were also immunoreactive with monoclonal antibody B72.3; however, a variety of other tumor types (including 13 melanomas) failed to stain. Staining by monoclonal antibody B72.3 was not noted in any of the 18 benign lesions aspirated, with the exception of occasional fine stippling in the cytoplasm of bronchial epithelial cells. Hence, monoclonal antibody B72.3 and fine needle aspiration biopsy techniques may be of potential use in the differential diagnosis and antigenic phenotyping of a spectrum of lung neoplasms prior to surgical resection.

Use of a monoclonal antibody (B72.3) as an immunocytochemical adjunct to diagnosis of adenocarcinoma in human effusions.

A monoclonal antibody (MAb), designated B72.3, has been generated using membrane-enriched fractions of a metastatic human breast carcinoma as the immunogen. Previous studies have demonstrated that the reactive antigen, a novel Mr 220,000 to 400,000 glycoprotein complex, can be detected in formalin-fixed, paraffin-embedded tissue sections of human breast and colon carcinomas, and not in a variety of normal adult human tissues. In this preliminary study, we report that MAb B72.3 may be used as an adjunct for diagnosis of adenocarcinoma in cytological preparations of human effusions. Using the avidin-biotin complex method of immunoperoxidase staining and formalin-fixed, paraffin-embedded cell suspensions, MAb B72.3 detected adenocarcinoma cells in effusions from all of 21 patients with adenocarcinoma of the breast. No reactivity was demonstrated in any cell type in benign effusions from 24 patients without cancer, or 13 patients with prior or extant cancer in other body sites; moreover, B72.3 showed no reactivity to leukemic or lymphomatous effusions, or apparent mesothelial cells from malignant effusions. MAb B72.3 also detected adenocarcinoma cells in cytological effusion specimens from 12 of 12 patients with adenocarcinoma of the lung and 16 of 16 patients with adenocarcinoma of the ovary. Thus, these data suggest that the immunocytochemical application of MAb B72.3 should now be considered as an adjunct in the discrimination of adenocarcinoma cells from reactive mesothelial cells in the cytological diagnosis of malignant effusions.

Adjunct to the diagnostic distinction between adenocarcinomas of the ovary and the colon utilizing a monoclonal antibody (COL-4) with restricted carcinoembryonic antigen reactivity.

Malignant ovarian tumors may represent either primary ovarian cancers or metastatic lesions (from patients with demonstrated primary cancers at other body sites) whose distinction may be difficult using clinical, surgical, and pathological criteria. Monoclonal antibody (MAb) COL-4, reactive with carcinoembryonic antigen, has previously been shown to react preferentially with adenocarcinomas of the colon versus a variety of normal tissues. We report here that MAb COL-4 is strongly reactive with primary colonic carcinomas (N = 50), as well as regional (N = 42), and distant (N = 20) metastases of colonic adenocarcinoma. In contrast, MAb COL-4 demonstrated little to no reactivity with primary (N = 53) and metastatic carcinomas of the ovary (N = 23) including serous, mucinous, and poorly differentiated adenocarcinomas using immunohistochemical techniques. This differential reactivity was statistically significant (P less than 0.001), suggesting the potential clinical utility of MAb COL-4 in the differentiation of ovarian from colonic adenocarcinoma. Solid-phase quantitative radioimmunoassays and Western blotting techniques confirmed these results. Data are also presented that the carcinoembryonic antigen molecules or epitopes recognized by a more classical broadly reactive anti-carcinoembryonic antigen MAb are distinct from those recognized by MAb COL-4. Other carcinomas which also metastasize to the ovary and may be confused clinically with a primary ovarian tumor such as adenocarcinomas of the stomach and breast were also evaluated for reactivity with MAb COL-4. COL-4 was also reactive with all gastric carcinomas evaluated, but failed to react with breast carcinomas. Hence, COL-4 can now be utilized as an immunohistochemical adjunct for the differentiation of ovarian from gastrointestinal adenocarcinoma which can be difficult to distinguish by clinical, surgical, and histological parameters.

Use of a monoclonal antibody (B72.3) as a novel immunohistochemical adjunct for the diagnosis of carcinomas in fine needle aspiration biopsy specimens.

Monoclonal antibody B72.3 has been shown to be reactive with a high-molecular-weight glycoprotein complex termed TAG(tumor-associated glycoprotein)-72. By the avidin-biotin immunoperoxidase method, fine needle aspirates and corresponding surgically excised tumor tissues from both malignant and benign tissues were analyzed for TAG-72 expression. Staining (range, 1 to 100 per cent of tumor cells) with monoclonal antibody B72.3 was observed in needle aspirates from 18 of 18 adenocarcinomas and adenosquamous carcinomas of the lung, 17 of 21 adenocarcinomas of the breast, and six of six adenocarcinomas of the colon, as well as adenocarcinomas from other body sites. In contrast, small cell carcinomas of the lung, malignant melanomas, lymphomas, and sarcomas did not stain with the antibody. Benign lesions from the breast, lung, pancreas, parotid, and thyroid also failed to stain. In 66 patients, tumor-bearing tissue had also been resected and was available for comparative examination with monoclonal antibody B72.3. In 62 of these 66 patients, the staining patterns in the aspirates were found to be predictive of the patterns of antibody reactivity in the comparable surgically resected tissues. From these studies it is concluded that monoclonal antibody B72.3 defines a tumor-associated antigen that is expressed in neoplastic cells but not in benign cells and is most selectively expressed in adenocarcinomas. This monoclonal antibody may be used as a novel adjunct for the diagnosis of carcinoma in fine needle aspiration biopsy specimens.

Epithelial ovarian tumors of low malignant potential.

The records of 31 women with ovarian tumors of low malignant potential were retrospectively reviewed to identify factors that determine the prognosis. Median follow-up was 51 months. Eighteen women had stage I disease. Twenty-three women (74%) had serous tumors, of which 46% were bilateral. Nine patients (29%) had concomitant endometriosis or endosalpingiosis. Two patients died of disease; both had mucinous tumors with extraovarian metastases at initial operation and inadequate pathologic sampling of their tumors. These results were combined with those of 970 women identified in previous reports to show that the rate of recurrence or persistence of ovarian tumors rises from 2% for women with stage I disease to 14% for those with stage III or IV disease, while mortality rises from 2 to 5%. Careful staging and pathologic sampling are important for establishing the prognosis. Testing of adjuvant therapy should be limited to patients with extraovarian disease.

Computed tomography-guided fine-needle aspiration of retroperitoneal lymph nodes in gynecologic oncology.

Computed tomography-guided fine-needle aspiration was performed on 30 retroperitoneal lymph nodes in 29 patients with gynecologic cancer. There were no serious complications. Review of the cytologic material demonstrated malignant cells in 83% of the aspirates. Because the predictive value of a positive aspirate approaches 100%, therapy for metastatic disease can be initiated in these patients with the need for an open biopsy. Among five aspirates in which malignant cells were not seen, the cellularity of the specimen appeared to be the critical factor determining the predictive value of the aspirate. Whereas neither of two patients with negative aspirates of adequate cellularity has developed recurrent disease, two of three patients with hypocellular negative aspirates have. Because a hypocellular negative aspirate from a retroperitoneal lymph node may not be a true reflection of disease status, either repeat aspiration or open biopsy is advisable.

Histologic features associated with hormonal responsiveness of ectopic endometrium.

OBJECTIVE: To correlate histologic parameters of endometriosis with hormonal responsiveness. DESIGN: Seven hundred sixty-eight unselected endometriotic implants and the corresponding intrauterine endometrium from 196 patients were classified by standard endometrial dating criteria. In addition, other histologic characteristics of endometriotic implants such as the amount of stroma, amount of fibrosis, the presence of surface epithelium, presence of focal hemorrhage, and gland characteristics were also noted. SETTING: Academic tertiary referral center. MAIN OUTCOME MEASURES: Comparison of histologic dating between endometriotic implants and the corresponding endometrium as a function of histologic parameters. RESULTS: Implants that were synchronous with the corresponding eutopic endometrium had more stroma than those that were out of phase. The amount of fibrosis was inversely related to hormonal responsiveness. The presence of surface epithelium in implants was also associated with an impaired response (28.0% versus 48.0% in phase). Endometriomas were found to be in phase with the corresponding endometrium less often than other types of implants (21.7% versus 43.3%). Although endometriomas had similar amounts of stroma when compared with other implants, they had significantly more fibrosis (850.2 microns versus 195.0 microns). CONCLUSIONS: These results suggest that the unpredictable response of endometriotic implants to cyclic endogenous hormones and hormonal therapy may be related to the architectural relationships between the cellular elements found in normal endometrium.

Cytomorphology of primary small-cell (Merkel-cell) carcinoma of the skin in fine needle aspirates.

The histologic appearance of primary small-cell carcinoma of the skin (the so-called Merkel-cell tumor) is similar to other small-cell tumors that may metastasize to the dermis. Significance has been placed on the electron microscopic appearance of this tumor since the ultrastructural features of this neoplasm are helpful in distinguishing it from most of the other neoplasms considered in the differential diagnosis. To determine whether any additional morphologic criteria might exist to distinguish this neoplasm, the fine needle aspirate appearance of a primary small-cell carcinoma of the skin was studied and compared to that of similar preparations of other small-cell tumors that could potentially involve the dermis. Cells of this unusual tumor were round and showed neither cohesiveness nor nuclear molding. Mitoses were numerous. The chromatin pattern was bland. The cytologic features of this tumor can aid in the distinction of primary small-cell carcinoma of the skin from other metastatic small-cell neoplastic lesions in the dermis of adults.

Fine needle aspiration of adenoid cystic carcinoma metastatic to the lung. Cytologic features and differential diagnosis.

Two examples of adenoid cystic carcinoma metastatic to the lung, one from a Bartholin's gland and the other from a submandibular gland, were sampled by fine needle aspiration. Although the cytologic features of adenoid cystic carcinoma have been well described, it is easy to confuse adenoid cystic carcinoma with other more common primary small-cell neoplasms of the lung; to determine distinguishing features, we compared the cytomorphology of adenoid cystic carcinoma with well-differentiated adenocarcinoma, small-cell undifferentiated carcinoma and carcinoid tumor of the lung. The differential features distinguishing adenoid cystic carcinoma from these other neoplasms include: (1) tight, globular, honey-comb arrangements of cells lacking true nuclear molding; (2) acellular chunks of basal lamina material, which alone may suggest adenoid carcinoma; and (3) the extension of a solid core of basal lamina material beyond a sievelike cellular meshwork. The morphologic expression of metastatic adenoid cystic carcinoma is so distinctive as to permit a definite diagnosis.

Patterns of reactivity of four novel monoclonal antibodies (B72.3, DF3, B1.1 and B6.2) with cells in human malignant and benign effusions.

The patterns of immunocytochemical reactivity produced in malignant and benign cells in effusions by staining with the monoclonal antibodies B72.3, DF3, B1.1 and B6.2 are reported. Using the mouse hybridoma methodology, these antibodies were raised against immunogens prepared from membrane-enriched extracts of metastatic carcinoma of the breast. Serial sections from the cell blocks of 20 benign effusions and 59 malignant effusions were incubated with the monoclonal antibodies using the avidin-biotin immunoperoxidase technique. The malignant neoplasms studied included specimens of 18 breast carcinomas, 12 ovarian carcinomas, 14 lung carcinomas, 7 leukemias/lymphomas and 8 miscellaneous malignant neoplasms. The monoclonal antibody B72.3 showed a positive staining reactivity with 36 of 39 adenocarcinomas (92%) versus staining in only 1 of 12 other neoplasms (8%). In contrast to the staining of adenocarcinomas, B72.3 stained no mesothelial cells in a total of 68 specimens tested. The monoclonal antibody DF3 showed a reaction pattern to neoplasms similar to that of B72.3 but also stained mesothelial cells in all of the specimens. The monoclonal antibodies B1.1 and B6.2 exhibited staining characteristics that have been associated with antibodies to carcinoembryonic antigen. The results of this study indicate that the monoclonal antibody B72.3 may function as a highly selective marker in recognizing a cancer cell versus a mesothelial cell and an adenocarcinoma cell versus other malignant tumor cells.

Prognostic value of cytologic examination of peritoneal washings in patients with endometrial carcinoma.

Peritoneal pelvic washings from 54 women with pathologic stage I endometrial carcinoma were evaluated in a blind retrospective fashion for the concentration of malignant cells present. None of the 42 patients with normal washings developed recurrence after a median disease-free survival of 36 months. Of the 12 patients with adenocarcinoma in the washings, 4 had high concentrations of malignant cells (greater than 1000 cells/100 ml sample), and all 4 died as a consequence of carcinoma within two years. The remaining eight patients had lower concentrations of malignant cells in the washings (less than 1000 cells/100 ml sample), and six of these patients had no evidence of disease after 37 to 64 months. Cox's nonparametric statistical model showed that increasing concentrations of adenocarcinoma cells in washings significantly shortened the time to recurrence of disease. The abundance of malignant cells has prognostic importance in identifying those patients with pathologic stage I disease who may require more aggressive therapy.

Detection of adenocarcinoma in peritoneal washings by staining with monoclonal antibody B72.3.

Peritoneal washings are routinely performed in the staging evaluation of carcinomas of the ovary and in "second-look" explorations; major problems in the evaluation of these specimens continue to be the distinction between atypical mesothelium and adenocarcinoma and the identification in an otherwise inflammatory specimen of rare cells of adenocarcinoma, which may be undetected by the most trained individual. Monoclonal antibody (MAb) B72.3, reactive with an oncofetal, tumor-associated glycoprotein (termed TAG-72; MW greater than 1000 kd) expressed in a variety of epithelial malignancies but not generally expressed in benign or malignant mesothelium, was reacted with sections from the paraffin-embedded cell blocks of 185 peritoneal washings from 180 patients with extant cancer or a prior history of malignancy. One hundred four of the washings were initially interpreted as atypical mesothelium, with no evidence of malignancy; when reacted with MAb B72.3, 6 of these specimens demonstrated groups of metastatic adenocarcinoma cells not appreciated by the usual cytologic criteria. Of the 81 washings interpreted as showing cells of adenocarcinoma, 73 demonstrated expression of TAG-72 from both gynecologic and nongynecologic malignancies. In the remaining 49 cases without an associated malignant process, MAb B72.3 did not stain atypical mesothelium, but did react, however, with benign endometrial cells and müllerian inclusions in two cases. MAb B72.3 may be used as a diagnostic adjunct to the routine cytologic evaluation of malignancy in peritoneal washings; reactivity with MAb B72.3 may indicate the need for further evaluation to define the presence of a malignancy or an advanced cancer.

Pigmented squamous cell carcinoma of the conjunctiva. A clinicopathologic ultrastructural study.

The clinical and morphologic features of a pigmented squamous cell carcinoma of the bulbar conjunctiva in a 57-year-old black man are reported. The tumor resembled the more usual epibulbar squamous cell carcinoma but had many melanin containing cells within it. Transmission electron microscopy disclosed melanosomes within squamous epithelium, melanocytes, and macrophages. The only cells within the tumor with morphologic features of a malignant neoplasm were squamous in type. The literature on this rare conjunctival tumor, which has a good prognosis following complete excision, is reviewed. Pigmented squamous cell carcinomas need to be distinguished from the more common nodular melanomas, which have a more ominous outlook.

Some observations concerning the demographic and geographic incidence of carcinoma of the lip and buccal cavity.

The geographic and demographic data obtained during the Third National Cancer Survey have provided a perspective on etiologic factors and incidence trends for cancers of low frequency. The incidence of cancer of the lip, oral cavity and skin from this survey was compared to similar studies in 1947 and intra-regional patterns in one area of the Third National Cancer Survey (Dallas-Fort Worth Metropolitan SMSA) were evaluated. The average age-adjusted annual incidence of cancer of the lip in white men in this latter area was 11.5 per 100,000 (based on 1950 population standard), 2-fold greater than that geographic area with the second highest incidence and approximately 3-fold greater than in all the other areas. The incidence in white women was only 8% that seen in white men. Intra-regional differences of similar populations were seen with the incidence in Fort Worth men being 50% greater than in a similar population in Dallas. Incidence trends over the past 2 decades reveal a significant decline in the incidence of oral cavity cancer and a slight decrease in lip cancer. Comparisons of the incidence of lip cancer did not correlate with the incidence of skin cancer nor with geographic latitude in the other survey areas. The studies fail to support the classical implication of actinic radiation as the primary etiologic factor in lip cancer incidence.

The presence of stage II melanosomes (premelanosomes) in neoplasms other than melanomas.

Aside from melanomas, other nonmelanocytic pigmented tumors synthesize melanin or contain benign passenger melanocytes. While Stage IV melanosomes (mature melanosomes) occur in neoplasms which synthesize melanin as well as in those with benign companion melanocytes, Stage II melanosomes (premelanosomes), which are found in melanocytes and cells of pigmented nonmelanocytic tumors of neural crest origin, are considered the morphologic hallmark of in vivo melanin synthesis. To test this widely held concept, we studied the ultrastructure of representative malignant melanomas and other pigmented tumors (pigmented variants of the nevocellular nevus, squamous cell carcinoma, schwannoma, basal cell carcinoma, and seborrheic keratosis). Discrete intracytoplasmic Stage II melanosomes were noted in neoplastic cells of tumors of neural crest origin (melanoma, schwannoma, and nevocellular nevus), which are widely believed to synthesize melanin. In addition, they were also detected in neoplastic epithelial cells of a squamous cell carcinoma, basal cell carcinoma, and seborrheic keratosis. In these epithelial tumors, a spectrum of melanosomes from Stage II through Stage IV were presumably acquired from nonneoplastic companion melanocytes, which were an integral part of the tumor. Because squamous epithelium has not been shown to synthesize melanin, this study suggests that the finding of intracytoplasmic Stage II melanosomes does not necessarily imply melanin synthesis. When accompanied by melanocytes, epithelial and perhaps other tumors may contain ingested Stage II melanosomes.