134

tracheal transplants 2, 4, 8, 12, and 16 weeks after cell inoculation proved to be of greater usefulness than either clinical or histological observation of the s.c. injection sites. A549, Calu-l, and TBE-I produced intratracheal neoplastic nodules as early as 2 weeks after cell inoculation. Calu-2, HUT292, and SW900 grew relatively slowly in the tracheas, and simple or stratified epithelia with slight or moderate atypia (preneoplastic lesions) were seen at 2 weeks. After the 4th week, they produced tumor nodules in the tracheal transplants, whereas no tumor cells could be seen at the s.c. injec- tion sites. The human derivation of the cells was confirmed by in situ hybridization using human-specific DNA probes. The cell lines offers a time- saving alternative to the s.c. inocula- tion assay for tumorigenicity and is at the same time a potentially valuable approach to studying preneoplastic and neoplastic progression with human cell subpopulations.

Morphological Features of Established Cultures of Human Squamous Lung Car- cinoma Cells and the Cellular Distribu- tion of Tumour-Specific Glycoproteins. Klokker, M., Jessen, H., Olsson, L., Behnke, O. Institute of Anatomy C, The Panum Institute, DK-2200 Copenhagen N, Denmark. Acta Pathol. Microbiol. Immunol. Scand. Sect. A Pathol. 94: 381-390, 1986.

Phenotypic characteristics of a cloned cell line RH-SLC-LII, estab- lished from a human squamous lung carcinoma, were studied. The line has maintained its morphologically charac- teristic growth pattern for over 3 years. Settling cells exhibited exten- sive surface blebbing during spreading and established small cell islands that eventually expanded by mitosis to con- fluent cultures. Cell islands and con- fluent cultures presented three cell types: (i) small, polygonal cells, (ii) polygonal cells of intermediary size and (iii) very large, extremely flattened, degenerating cells. Mitotic activity was present predominantly in type (i) and the sequence (i) - (iii) is presumed to represent the lines' cycle. Previous work has demonstrated that the SLC-LII line releases tumor- associated glycoproteins and glycolipics. These could be identified with a murine Mab (43-9F). The specific epitope was determined by carbohydrate residues and was shown to have growth factor-like properties. Mab 43-9F bound heterogeneously to the surface of SLC- LII cells: Most large cells were un- reactive while both type (i) and (ii) cells showed conspicuous differences in immunostaining intensity. Im- munocytochemical analysis also indi-

cated redistribution, shedding and in- ternalization of antigen-Mab complexes, which may have significant impact on the use of the epitope as tumor marker in diagnosis and therapy. No definite clue was obtained as to the release of the antigenic carbohydrate epitope itself.

Reduced in Vitro Erythroid Progenitor Cell Growth in Bronchial Cancer. Masters, G.S., Baines, P., Bailey-Wood, R. et al. Department of Haematology, University of Wales College of Medicine, Cardiff, U.K. J. Clin. Pathol. 40: 87-93, 1987.

Peripheral blood and bone marrow were studied in 21 men with dissemi- nated untreated bronchial cancer in an attempt to define abnormalities of erythropoiesis associated with the development of anaemia. Haemoglobin concentratin at or below 13 g/dl was present in 13 cases. Marrow morphology was normal in all cases except one, in which small number of tumur cells were found. Clonal assay of erythroid progenitors showed a significant decrease in the number of BFU-E (p = 0.03) and CFU-E (p = 0.01) compared with cultures from normal marrow (12 subjects). The growth of granulocyte and macrophage progenitors (GM-CFCs) was similar in patients with bronchial cancer and normal subjects. When normal marrow was incubated in the presence of serum from bronchial cancer patients, no inhibitory factors could be detected either for BFU-E or CFU-E growth. In all patients circulating T8 numbers were significantly raised (p = 0.0002). Consequently, the median T4:T8 ratio in blood was 1.2, and this was sig- nificantly lower than the ratio of 1.7 found in 20 normal subjects (p = 0.036). In 18 patients the bone marrow T4:T8 ratio of i.i was significantly lower than the ratio of 2.9 found in seven normal subjects (p = 0.04). Total blood white cell counts, neutrophils, and monocyte numbers were also in- creased (p = 0.0001; p = 0.0001; p = O.002).

Two Murine Monoclonal Antibodies Against Human Lung Cancer-Associated Antigens. Endo, K., Kamma, H., Ogata, T. Depart- ment of Surgery, Institute of Clinical Medicine, Ibaraki-ken 305, Japan. Can- cer Res. 46: 6369-6373, 1986.

Two murine monoclonal antibodies, MAb 8 (immunoglobulin Glkappa), were produced after immunization with TKB-2, a variant cell line of human small cell carcinoma of the lung. In enzyme-linked immunosorbent assay, these antibodies reacted with four major types of lung cancer cell lines and various extrapul- monary tumor cell lines. Immunohis-

tological study, however, showed highly specific binding to lung cancers; MAb 8 bound to 68% of 65 lung cancers, and MAb 15 bound to 72% of them. Interestingly, both antibodies were more reactive with non-small cell than small cell lung cancers and bound most frequently to large cell carcinoma. Most extrapulmonary tumor tissues were negative in staining with a few exceptions; endodermal sinus tumor (two of two) was positive to both antibodies, breast carcinoma (one of five) to MAb 8, gastric carcinoma (one of three), and malignant melanoma (one of one) to MAb 15. Cross-reactions with normal tissues were limited; MAb 8 reacted with adult and fetal lung, and MAb 15 with esophagus and renal tubules. MAb 8 recognized a M(r) 48,000 glycoprotein antigen (carbohydrates as its epitope), and MAb 15 recognized two proteins (M (r) 85,000 and 45,000) (peptides as their epitopes). These two antibodies detecting novel antigens ex- tensively associated with an highly specific to lung cancers, are poten- tially useful for the study of lung cancer.

4. PATHOLOGY

Histopathology of Lung Cancer in New Mexico, 1970-72 and 1980-81. Butler, C., Samet, J.M., Humble, C.G., Sweeny, E.S. Department of Pathology, University of New Mexico School of Medicine, Albuquerque, MN 87131, U.S.A. J. Natl. Cancer Inst. 78: 85-90, 1987.

In conjunction with a population- based case-control study of lung cancer in New Mexico, the histopathology of cases diagnosed durign 1980 and 1981 and during 1970-72 was reviewed. Adequate histologic or cytologic material was obtained for 725 cases, with 308 during 1970-72 and 417 during 1980-81. The light microscopic his- tologic type was classified on the basis of review by 2 pathologists. No significant differences were found in the histologic-type distributions in Hispanics and non-Hispanics whites. In males, the distributions of histologic types were similar in the two time peirods, but in non-Hispanic white women the proportion of adenocarcinoma declined.during 1980-81 as the propor- tion of small cell carcinoma increased. The panel classification was compared with that recorded by the New Mexico Tumor Registry. Overall agreement was 52.1% for 1970-72 and increased to 65.2% for 1980-81. The discrepancies between the two classifications were largest for the categories of large cell undifferentiated carcinoma and 'other malignancy'

135

Immunohistological Staining of Reactive Mesothelium, Mesothelioma, and Lung Carcinom~ With a Panel of Monoclonal Antibodies. Ghosh, A.K., Gatter, K.C., Dunnill, M.S., Mason, D.Y. Nuffield Department of Pathology, John Radcliffe Hospital, Oxford, U.K.J. Clin. Pathol. 40: 19- 25, 1987.

A panel of seven monoclonal an- tiepithelial antibodies of different specificities, including anticytoke- ratin, human milk fat globule membrane, Ca, and carcinoembryonic antigen (CEA) were used with the alkaline phosphatase-antialkaline phosphatase (APAAP) immunostaining technique to determine their value in the differen- tiation between benign and malignant mesothelial cells and lung carcinoma in histological preparations. The an- ticytokeratin antibody reacted strongly with all cases of reactive mesothelium, mesothelioma, and lung carcinoma. An- tibodies to human milk fat globule membrane and the Ca antigen stained mesothelioma and carcinoma and 43% of cases of reactive mesothelium. Staining for carcinoembryonic antigen was not detected in reactive mesothelium or mesothelioma, but was present in most of the lung carcinomas. CEA seemed to be the single most useful marker in distinguishing carcinoma from mesothelioma in that a positive reac- tion for CEA would indicate carcinoma rather than mesothelioma.

Limitations of the Usefulness of Microvillous Ultrastructure in Distin- guishing Between Carcinoma Primary in and Metastatic to the Lung. Engstrand, D.A., England, D.M., Oberley, T.D. Middleton Memorial Veterans Hospital, Madison, WI 53705, U.S.A. Ultrastruct. Pathol. ii: 53-58, 1987.

We performed ultrasound analysis on 70 consecutive patients with solitary cancers in lung with the following his- tologic classifications: adenocarcinoma (42 cases), bronchioloalveolar car- cinoma (13), large cell carcinoma (4), and adenosquamous carcinoma (ii). Off these 70 cases, nineteen (13 adenocarcinomas, 4 bronchioloalveolar carcinomas, and 2 adenosquamous carcinomas) contained cell surface microvilli with microvillus core root- lets and/or glycocalyceal bodies. Sub- sequent clinical follow-up revealed that three of these 19 cases were ac- tually metastatic colon carcinomas. The remaining 16 patients are currently free of extrathoracic primary disease and are therefore, presumably, primar M carcinoma of the lung. Since both primary and metastatic tumors showed cell surfaces with microvilli having core rootlets and glycocalycean bodies,