Mucoepidermoid carcinomas: immunohistochemical studies on keratin, S-100 protein, lactoferrin, lysozyme and amylase

Immunohistochemical expression of 8 cases of mucoepidermoid carcinomas (G-I, 3 cases; G-II, 2 cases; and G-III, 3 cases) revealed marked heterogeneity of the proteins examined. Immunohistochemically detectable keratins (TK, KL1, and PKK1) were distributed in epidermoid cells, but were absent in mucous secreting cells. Strongly positive deposits of keratin proteins were detected in squamoid tumor cells in the G-I tumors. The tumor cells displayed positive staining for S-100 alpha, but did not stain with polyclonal S-100 antiserum or with monoclonal S-100 beta. The cells showing highest reactivity for S-100 protein were scattered in neoplastic foci and were probably Langerhans cells. Lactoferrin and lysozyme reactions were generally negative in tumor foci; but a positive reaction for lactoferrin was found in luminal tumor cells although rarely, and lysozyme staining was occasionally noted in histiocytes in the stroma. Amylase activity was usually absent in the tumor cells, with the exception of one case in which it was confined to the tumor cells. Mucoepidermoid carcinomas of various grades indicated marked heterogeneity in terms of various immunohistochemically detectable proteins.     

Malignant ascites of serous papillary ovarian adenocarcinoma. An immunocytochemical study of the tumor cells

In 17 malignant peritoneal effusions due to papillary serous adenocarcinoma of the ovary, the reaction patterns of the tumor cells to monoclonal antibodies (MAbs) against surface antigens were studied and compared with the reaction patterns of mesothelial cells in the same effusions. The following surface markers were used with the adhesive slide method: epithelial membrane antigen (EMA), human epithelium-specific cell surface antigen (HEA-125), human endothelial antigen (BMA-120), carcinoembryonic antigen (CEA 3-13), an antibody against natural killer cells and cytotoxic cells (BMA-070), granulocyte antigen (Leu M1) and leukocyte antigen of class I (HLA-1). In all cases, from 30% to 95% of the tumor cells reacted with EMA and HEA-125. Tumor cells showed a positive staining with CEA 3-13 in only five cases. In all cases, from 75% to 95% of the tumor cells reacted positively with BMA-120. The reactivity of a few mesothelial cells with EMA and of all mesothelial cells with BMA-120 did not interfere with the identification of positive tumor cells since the reaction patterns were different. Interestingly, our study demonstrated that BMA-070, an MAb identifying natural killer cells and cytotoxic cells, is also a most useful tumor marker. The same was found to be true for Leu M1, an MAb originally thought to react only with granulocytes. The tumor cells showed a partial or total loss of the expression of HLA-1 reactivity. Since all cases were immunocytochemically positive for tumor cells while conventional cytology was positive in only 13 of the cases, the immunocytochemical analysis of malignant peritoneal effusions due to papillary serous adenocarcinoma of the ovary seems able to improve the cytologic diagnosis of the fluids.     

Malignant pleural effusions due to small cell carcinoma of the lung. An immunocytochemical cell-surface analysis of lymphocytes and tumor cells

Thirteen malignant pleural effusions due to small cell carcinoma (SCC) of the lung were immunocytochemically studied using the peroxidase-antiperoxidase adhesive slide assay for the determination of cell surface antigens. A panel of monoclonal antibodies (MAbs) was used to determine the lymphocyte subpopulations and the reactivity of the tumor cells. Of the lymphocytes, 87 +/- 1% were CD3+ T cells, with 72 +/- 10% CD4+ helper/inducer T cells and 20 +/- 5% CD8+ suppressor/cytotoxic T cells. Only a minority of T lymphocytes were activated in terms of expressing the surface markers CD38 and HLA-DR. The distribution of the lymphocyte subpopulations was not significantly different from the distribution in other malignant and nonmalignant pleural diseases previously studied, indicating that the reaction pattern of the lymphocytes in the pleural cavity is similar in different diseases. The tumor cells from all cases were positive for LeuM1, CD16 and HLA-DR; 10 of 11 cases were positive for HEA-125, Sam 2 and Sam 10. Positivity for epithelial membrane antigen was observed in 11 cases, for OKT9 in 8 cases and for carcinoembryonic antigen in 6 cases. A total or partial loss of the reactivity with HLA-1 was found in nine cases. The reactivity pattern of the tumor cells with the MAbs used in this study is not specific for SCC of the lung because other carcinoma cells also reacted with these markers. Additional morphologic criteria, such as cell size and cell configuration, are needed to recognize the immunocytochemically positive-reacting cells as tumor cells from SCC of the lung. However, the immunostaining allows a better identification of the tumor cells, especially in cases with a small quantity of tumor cells.     

The comedo subtype of intraductal carcinoma. Cytologic characteristics.

Smears from 10 intraductal carcinomas of the comedo type without microinfiltration were compared with smears from 10 similar tumors with suspected or proven microinfiltration and smears from 10 invasive comedo carcinomas. Microinvasive tumors could not be separated from purely intraductal cases unless tumor cell infiltration in mammary fat was seen in the smear. The tumor cells in most of the intraductal cases were shed in cohesive groups and clusters, lying in necrotic cellular debris and with few or no scattered, single, dissociated tumor cells. Smears from invasive comedo carcinomas invariably showed tumor cell clusters and scattered, single, dissociated tumor cells, often with atypia in excess of what was seen in the intraductal cases. Also, in most cases the invasive tumors did not show a background of necrotic cellular debris.     

Selective loss of HLA-A or HLA-B antigen expression in colon carcinoma

A panel of colorectal carcinomas has been examined immunohistologically with mAb specific for allodeterminants of HLA-A or HLA-B, and with mAb reactive with HLA-A,B,C framework determinants or beta 2-microglobulin. Out of 85 carcinomas, 5 cases were completely negative for all class I Ag in the tumor cell population. Fifteen cases exhibited a differential expression of HLA-A and HLA-B products in all tumor cells or in a subset. Such tumor cells showed strong staining with monomorphic HLA antibodies but failed to stain with certain allospecific mAb. Negativity of tumor cells was scored only when normal stromal cells within the tumor mass were clearly labeled, indicating expression of the particular allodeterminant(s) in the given tissue. Four carcinomas showed a selective loss of HLA-A2 or a non-A2 HLA-A allospecificity in all tumor cells or a major subset, whereas HLA-B Ag were expressed. Seven cases showed a selective loss of the HLA-Bw6 superspecificity in the tumor cell population; HLA-Bw4 and HLA-A Ag were present. Three cases exhibited a combined loss of HLA-A2 and HLA-Bw6 Ag, with non-A2 HLA-A and HLA-Bw4 Ag being expressed. A further case was HLA-Bw6/Bw4 negative in a major tumor cell subset and HLA-A negative in the complementary minor subset. The results show that a considerable proportion of colorectal carcinomas is heterogeneous with regard to HLA-A and HLA-B expression. The reasons for the appearance of tumor subsets with complete loss of class I Ag or selective loss of only HLA-A or HLA-B Ag are not clear, but it is conceivable that some of them arose because they have escaped immunoselection.     

Diagnosis of human childhood rhabdomyosarcoma of antibodies to desmin, the structural protein of muscle specific intermediate filaments

Four embryonal rhabdomyosarcomas, one tumor diagnosed as an undifferentiated sarcoma, probably a rhabdomyosarcoma, and six different non-muscular sarcomas were investigated with antibodies specific for different intermediate filament types. The tumor cells in the rhabdomyosarcomas and the undifferentiated tumor were stained clearly by antibodies to desmin, the intermediate filament type characteristic of muscle. The staining of tumor cell by antibodies to vimentin, the intermediate filament type characteristic of certain cell types of mesenchymal origin including myoblasts, was different in these 5 cases. In one case of embryonal rhabdomyosarcoma nearly all tumor cells were stained, but in the remaining cases few or no tumor cells were positive with the vimentin antibody. In these rhabdomyosarcomas not only the large rhabdomyoblasts, but also the small undifferentiated cells were labeled by antibodies to desmin. In the latter cell type the desmin filaments were arranged typically in coils. In contrast, tumor cells in the non-muscular mesenchymal sarcomas were stained only by antibodies to vimentin but not by antibodies to desmin or prekeratin. The retention of the desmin marker characteristic of normal muscle in cases of rhabdomyosarcoma not only allowed the undifferentiated desmin-positive sarcoma to be classified as rhabdomyosarcoma but also suggests that the use of antibodies to desmin could be very helpful in the future for the diagnosis of undifferentiated rhabdomyosarcomas.     

The value of the immunoperoxidase slide assay in the diagnosis of malignant pleural effusions in breast cancer

Whether immunocytochemical studies of malignant pleural effusions due to breast cancer would increase the diagnostic yield as compared with conventional effusion cytology was examined in 30 cases with biopsy-proven metastatic spread to the pleura. Conventional cytology was performed on air-dried smears as well as on cytocentrifuge preparations stained with the May-Grünwald-Giemsa stain. Immunocytochemistry was performed with monoclonal antibodies against carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA) and human leukocyte antigen (HLA) and the peroxidase-antiperoxidase technique on glass slides after Ficoll-Hypaque centrifugation. By conventional cytology, 13 cases (43%) were positive for malignant cells, 6 cases (20%) were suspicious, and 11 cases (37%) were negative. In marked contrast, all 30 cases were immunocytologically positive for malignancy. Tumor cells in all cases demonstrated a positive reaction for EMA. Some mesothelial cells were also positive for EMA, but their reaction pattern was clearly distinguishable from that of the tumor cells. Twenty-one cases (70%) also showed CEA-positive tumor cells; mesothelial cells never reacted with CEA. Some tumor cells showed a loss of HLA expression. In conclusion, this immunocytologic method can be recommended as a routine procedure for greatly increasing the diagnostic yield of cytology in pleural effusions due to breast cancer.     

Hemangioendothelioma. Cytologic appearances in two cases presenting with multiple soft tissue and bone lesions

Two cases of hemangioendothelioma were detected by fine needle aspiration cytology in two women presenting with multiple soft tissue and bone lesions. The aspirates were hemorrhagic, and the smears showed mostly dissociated and predominantly monomorphic tumor cells. Focal rosettelike clustering of tumor cells was also present. A prominent cytologic feature of these two cases of hemangioendothelioma was the close association of tumor cells with endothelium-lined vascular structures, a pattern that can be seen in other sarcomas and sometimes in malignant epithelial tumors.     

Alveolar rhabdomyosarcoma of the vulva. Report of two cases

The cytologic and histologic findings in two cases of the extremely rare alveolar rhabdomyosarcoma of the vulva are reported. The tumor cells in fine needle aspiration smears from one case and tumor imprints from both cases were isolated or were in sheets or reticular patterns. The nuclei were round to oval; only a few cells were multinucleated. The chromatinic material was increased in amount and finely granular. Many mitotic figures were observed. The cytoplasm was scanty in general, but some cells had abundant cytoplasm; cross striations were not recognized. The tumor cells were positive with immunocytochemical stains for desmin, vimentin and myoglobin. Similar findings were observed in biopsy and surgical specimens.     

Cathepsin D expression in colorectal adenocarcinomas and adenomas

The aim of this study was to investigate the role of cathepsin D in colorectal cancer. For this purpose cathepsin D expression was evaluated by means of immunohistochemistry in stromal and tumor cells of 31 colorectal carcinomas and 29 adenomas. Cytoplasmic cathepsin D expression of tumor cells was present in 90.3% of the carcinoma cases and various degrees of stromal cell cathepsin D expression were present in all cases. In the adenomas, the epithelial cells and stromal cells expressed cathepsin D in 68.96% and 96.55% of cases, respectively. The staining intensity was always weaker in the adenomas. When the stromal and tumor cell cathepsin D expression in the adenocarcinoma and adenoma cases were compared, a statistically significant difference was observed in the staining of stromal cells. Furthermore, stromal cathepsin D expression in the adenocarcinomas was related to tumor stage when the carcinomas were divided into low and high stage. Cathepsin D expression in stromal cells may be an important indicator of poor prognosis in colorectal adenocarcinomas.     

Cytopathologic aspects of leiomyoblastoma in fine needle aspiration biopsy. Report of two cases

The cytopathologic features of the fine needle aspiration biopsy of two cases of malignant peripheral leiomyoblastoma are presented. Aspirated tumor cells occurred singly, not in clusters. They were pleomorphic, varying in size and shape and showing eosinophilic to amphophilic cytoplasm. Many tumor cells had a vacuolated cytoplasm. Multinucleated large tumor cells were seen in one case; their nuclei were hyperchromatic and round to oval, with chromatin clumping and prominent nucleoli. The cytologic findings suggested a malignant mesenchymal tumor. Histology demonstrated the diagnosis of leiomyoblastoma.     

Comparison of CEA distribution in lesions and tumors of salivary glands as determined with monoclonal and polyclonal antibodies

Immunohistochemical localization of carcinoembryonic antigen (CEA) with conventional antibody to CEA (anti-CEA), nonspecific crossreacting antigen (NCA)-absorbed polyclonal antibody to CEA (NCAa-CEA), and monoclonal antibody to CEA (Mono-CEA) have been compared in obstructive lesions and salivary gland tumors. Normal salivary glands gave strong staining of the luminal borders of acinar cells with anti-CEA, whereas no staining occurred with Mono-CEA. Obstructive lesions showed occasionally marked staining with anti-CEA in some acinar cells, but there was no reaction with Mono-CEA. Of 69 pleomorphic adenomas examined, 34 were positively stained with anti-CEA, 18 with NCAa-CEA and 8 with Mono-CEA along the luminal borders of the tumor cells. The frequency of positive staining of material within tubular lumina was similar with all three immunoreagents. Neoplastic cells were positive with Mono-CEA in only three cases, while eight cases were positive with NCAa-CEA and 11 cases with anti-CEA. In salivary gland tumors true CEA is be found mainly at the border of tumor cells, but the frequency of positive reactions is low.     

Proliferative response of lymphocytes from ovarian cancer patients to autologous tumor cells

Summary Peripheral blood lymphocytes (PBL) from 43 patients with histologically confirmed ovarian carcinoma were stimulated in mixed lymphocyte-tumor culture (MLTC) with purified autologous tumor cells. Positive results, assessed as lymphocyte proliferation, were observed in 21 cases (48.8%). Lymphoid cells associated with ascitic fluid or infiltrating solid masses were in general less reactive than PBL as only 3/11 cases had positive MLTC. Tumor cells isolated from peritoneal effusions showed no significant difference in stimulatory potential as compared to the primary tumor. These results suggest that in an appreciable proportion of ovarian carcinoma patients (approximately 50%), lymphocytes have the potential to react to autologous tumor cells. Comprehension of the immunological mechanisms of antitumor resistance may have direct practical relevance for more effective treatment of neoplasms.     

Fine needle aspiration cytopathology of phyllodes tumor. Differential diagnosis with fibroadenoma

The differential diagnosis of benign or borderline phyllodes tumors of the breast in fine needle aspiration biopsy smears was studied. Smears from five histologically proven cases and 20 proven cases of fibroadenoma were evaluated with regard to their cytologic features. The findings indicate that the presence of a high cellularity of stromal fragments (including bipolar naked nuclei), clusters of hyperplastic ductal cells and giant cells and the absence of apocrine metaplasia may suggest a diagnosis of phyllodes tumor, as opposed to fibroadenoma. The borderline phyllodes tumor showed a sarcomatous atypia and/or small intranuclear cytoplasmic invaginations in some cells. The differential diagnosis of this tumor with malignant phyllodes tumor and plasma-cell mastitis is also discussed.     

Tumor structure and extracellular matrix as a possible barrier for therapeutic approaches using immune cells or adenoviruses in colorectal cancer

In this article we report about the role that tumor structure and extracellular matrix (ECM) may play in immunotherapy and in gene therapy using adenoviruses. We performed studies in a rat model for colorectal cancer, CC531, and in specimens of human colorectal cancer. The tumors were composed of two compartments, tumor cell nests surrounded by stromal cells. ECM proteins were expressed in the stromal part, where the blood vessels were also located. Furthermore, in several tumors, the tumor cell nests were surrounded by basal membrane-like structures. Therefore, in vascular approaches to treat cancer, therapeutic agents on their route to tumor cells may be hampered by ECM to reach tumor cells. We found that immune cells were abundantly present in tumors from colorectal origin. These cells were, however, not found in direct contact with tumor cells, but mainly in the stromal part of the tumor. Adenoviruses, when intravascularly injected, did not reach tumor cells in the CC531 rat model. Tumor cells were only infected, and even then in limited numbers, in cases of intratumoral injection. We hypothesize that ECM in a tumor is a barrier both for immune cells and for adenoviruses to make direct contact with these tumor cells, and thus limits colorectal tumor therapy.     

Bone metastases: prognostic applications of cytometric DNA analysis

OBJECTIVE: To examine DNA parameters as prognostic factors for developing metastases. STUDY DESIGN: Image cytometry was used to determine DNA content of 21 tumors and 28 metastases. DNA ploidy status, 2c deviation index (2cDI) and DNA malignancy grade (DNA-MG) (based on the variation of nuclear DNA content of tumor cells around the normal DNA [2c] peak) were examined for their prognostic value. RESULTS: Twenty of 21 tumors showed aneuploid content, and 1 tumor showed diploid DNA content. Twenty-one bone metastases showed aneuploid cells. In 6 cases both euploid and aneuploid cells were detected. In 1 metastasis only euploid cells were present. DNA-MG was increased in bone metastases (mean, 2.4) as compared to the corresponding primary tumor (mean, 2.2) in most of the cases. The mean value of the 2cDI was 30.07 in primary tumors and 42.5 in metastases. Twelve bone metastases had a higher 5cEE than did the primary tumor. CONCLUSION: Diploid and aneuploid cells were able to leave a tumor and establish metastases. DNA-MG and 2cDI were increased in metastases in comparison with the primary tumor, but even tumors with lower DNA-MG had metastatic potential.     

Fine needle aspiration cytology and immunocytochemistry of large cell lymphomas expressing the Ki-1 antigen

Cytomorphology and immunocytochemistry were used to diagnose two cases of large cell anaplastic lymphomas that expressed the Ki-1 antigen. Smears stained with the May-Grünwald-Giemsa stain showed a cytomorphologic spectrum that varied from immunoblastlike cells to large tumor cells with pleomorphic nuclei. The tumor cells had a distinct basophilic cytoplasm, which often contained vacuoles. Giant cells with wreathlike multilobulated bizarre nuclei were also observed. Immunocytochemistry on a cytocentrifuged specimen showed that the tumor cells were of a T-helper phenotype; all strongly expressed the Ki-1 antigen and receptors for HLA-DR and IL-2.     

Immunomorphological study of 1,2-dimethylhydrazine-induced carcinomas of the large intestine in rats using monoclonal antibodies to intermediate filament proteins

Cryostatic sections of rat large bowel tumors induced by 1,2-dimethylhydrazine were stained with monoclonal antibodies against different proteins of intermediate filaments: (a) against prekeratin (mol. mass 49 000, PK49) found in many epithelial cells and (b) against vimentin, a constituent of intermediate filaments of mesenchymal cells. Immunofluorescence study showed that large bowel tumor cells as well as normal cells of this organ contain PK49 but not vimentin. High sensitivity of the method allowed one to clearly identify small invasive nodules and groups of tumor cells not visible in usual histologic preparations. Moreover, in some cases single atypical tumor cells were identified in tumor stroma and in the submucosal layer underlying the tumor, that were indistinguishable from normal mesenchymal cells at the light microscopy level.     

Metastatic breast carcinoma in cerebrospinal fluid. A cytomorphometric study

A study was undertaken to quantitate the cellular characteristics of metastatic breast carcinoma in cerebrospinal fluid (CSF). Millipore filters of CSF from 15 patients with metastatic breast carcinoma were reviewed; 50 cells per case were evaluated when available. All cells in all cases shed singly or in loose clusters; tight balls or morulae were absent. All cells had regular, round-to-oval nuclei with finely granular chromatin. The majority of cells in all cases had single or multiple round nucleoli, granular cytoplasm with distinct borders and a mean nuclear-cytoplasmic ratio of close to 0.70. Cellular background, number of tumor cells per case, number and placement of nuclei and nuclear and cytoplasmic diameter varied both within and among the cases. There was significant variation in nuclear and cytoplasmic diameters both within and among the cases of infiltrating ductal carcinoma. Thus, the uniform appearance of the cells was due to consistent cytologic features, not to similarity in cell size. The cytologic profile of metastatic breast carcinoma is sufficiently characteristic to distinguish this tumor from other benign and malignant lesions that shed in the CSF.     

Fine needle aspiration cytology of the Ewing's sarcoma family of tumors

OBJECTIVE: To study the cytomorphologic features of the Ewing's sarcoma (ES) family of tumors. STUDY DESIGN: During a period of eight years (1990-1997), 123 soft tissue tumors and 65 bone tumors were evaluated by fine needle aspiration cytology (FNAC); 14 cases were diagnosed as in the ES family of tumors. The ages of the patients ranged from 8 to 30 years. All the cases were histologically confirmed. RESULTS: Of 14 cases of the ES family of tumors, 7 were ES, 3 extraosseous ES (EOE), 2 peripheral primitive neuroectodermal tumor (PPNET) and 2 Askin tumor. Cytologically, smears from all the cases showed round tumor cells with a high nuclear/cytoplasmic ratio. On detailed examination, subtle differentiating features were observed. The cells in ES had finer nuclear chromatin in comparison to those of PPNET and Askin tumor, and punched-out clear cytoplasmic vacuoles were present. PPNET showed nuclear molding, unipolar cytoplasmic tags and Homer-Wright rosettes. Histologically, all cases of ES and EOE and one case of Askin tumor showed periodic acid-Schiff-positive inclusions. CONCLUSION: FNAC features coupled with clinical findings enable a rapid diagnosis of the ES family of tumors, from which treatment modalities can be determined.