An immunohistochemical study of lung carcinomas using a monoclonal antibody which specifically reacts to basal cells

Sixty-three primary lung carcinomas were examined immunohistochemically using a monoclonal antibody, KP8D4, which reacted specifically to basal cells in the lung. Seven of ten squamous cell carcinomas, four of 41 adenocarcinomas, two of five large cell carcinomas, and one adenosquamous carcinoma were reactive with KP8D4; however, four small cell carcinomas, one mucoepidermoid carcinoma, and one carcinoid tumor were all not reactive to this antibody. Four adenocarcinomas stained by KP8D4 were Clara cell type adenocarcinomas, and two of them also revealed reactivity to surfactant apoprotein in the same cells labeled with KP8D4. These results indicate that carcinomas positive to KP8D4, including adenocarcinomas at the lung periphery, expressed phenotypically the same protein which was specifically recognized in the basal cells, suggesting that the basal cells possessing a multipotent capability of differentiation are a possible candidate of progenitor cells of lung carcinomas, including peripheral type lung carcinomas.     

Clinicopathologic and Immunohistochemical Characteristics of Goblet Cell Type Adenocarcinoma of the Lung

Twenty-two resected goblet cell type adenocarcinomas of the lung were examined clinicopathologically and immunohistochemically. The stage and survival curve of goblet cell type adenocarcinomas were compared with those of 44 cases of pure or mixed Clara cell and bronchial surface epithelial cell (Clara and BSE) type adenocarcinomas. Each case of goblet cell type was matched with two cases of Clara and BSE type as to sex, age and date of surgery. In goblet cell type adenocarcinomas, lymph node metastasis was less frequently and intrapulmonary metastasis was more frequently detected than in other types of adenocarcinomas (p < 0.001 and p < 0.05, respectively). Goblet cell type adenocarcinomas showed better prognoses than Clara and BSE type adenocarcinomas. However, the estimated survival curves of those two groups become similar after adjustment of the TNM condition using Cox's proportional-hazard general linear model. This result indicated that the longer survival of goblet cell type adenocarcinoma was due to the characteristic distribution of TNM conditions, that is, unique local growth and low incidence of lymph node metastasis. When goblet cell type adenocarcinoma was macroscopically classified into two types, i.e. solitary peripheral nodule type (nodular type) and multifocal nodular type or consolidation of all or part of a lobe (diffuse type), the nodular type had better prognosis than the diffuse type (p < 0.05). Immunohistochemically, 83%, 11%, and 0% of goblet cell type adenocarcinomas were positive for NCC-CO-450, carcinoembryonic antigen (CEA), and surfactant apoprotein, respectively. Most Clara and BSE type adenocarcinomas were negative for NCC-CO-450, but positive for CEA and surfactant apoprotein. NCC-CO-450 was considered to be a good immunohistochemical marker of goblet cell type adenocarcinoma of the lung. These results indicated that goblet cell type tumors are different from most adenocarcinomas of other types both clinicopathologically and immunohistochemically. Acta Pathol Jpn 41: 737-743, 1991.     

Expression of surfactant associated protein-A and Clara cell 10 kilodalton mRNA in neoplastic and non-neoplastic human lung tissue as detected by in situ hybridization.

Two markers for the progenitor cells of peripheral airways and their tumors are the 10 kilodalton (kd) Clara cell protein and the major surfactant associated protein-A (SP-A). We used the RNA-RNA in situ hybridization technique to study expression of the genes encoding these proteins at the cellular level in 19 pairs of non-neoplastic and neoplastic tissues from resected human lungs. Our results show that in non-neoplastic lung tissue, the Clara 10 kd protein gene was expressed in nonciliated cells of both bronchial and bronchiolar epithelium, indicating that, in contrast to previous assumptions, cells with Clara cell-like differentiation in humans may not be restricted to bronchiolar cells. The incidence of Clara 10 kd protein gene expression, as detected in lung carcinomas (1 out of 19 cases positive) was less than expected based on previous ultrastructural reports. The SP-A gene was strongly expressed in normal alveolar type II cells in non-neoplastic lung and, at higher levels, in hyperplastic cells. In addition, SP-A mRNA expression was observed in scattered bronchial and bronchiolar epithelial cells in 40% of the airways examined. Five out of 17 lung tumors, all of which were adenocarcinomas, were positive for SP-A expression, albeit generally less intense than type II cells. This expression was seen in carcinomas with papillolepidic as well as solid and glandular growth patterns. Our findings provide new insights into the peripheral airway cell differentiation.     

The value of PE-10, a monoclonal antibody against pulmonary surfactant, in distinguishing primary and metastatic lung tumours

A new monoclonal antibody (PE-10) raised against components of pulmonary surfactant has been assessed for its ability to distinguish primary from secondary carcinomas in the lung. We applied this antibody to a series of 107 primary lung carcinomas, 40 adenocarcinomas of other sites, and 26 cases of adenocarcinoma metastatic to lung and pleura. Of the primary lung carcinomas, all the non-mucinous bronchiolo-alveolar carcinomas were positive whereas all the mucinous cases were negative; 60% of other types of adenocarcinoma were positive and 10% of large cell undifferentiated carcinomas, 20% of small cell carcinomas and 40% of atypical carcinoids also showed focal positivity. Squamous cell carcinomas were all negative. Adenocarcinomas of the breast, kidney, large bowel and ovaries were all negative, as were all 26 cases of adenocarcinoma metastatic to the lung and pleura. We conclude that this antibody is highly specific and moderately sensitive for primary tumours of the lung.     

Immunohistochemical studies on the expression of pulmonary surfactant apoproteins in human lung carcinomas using monoclonal antibodies

To reevaluate the expression of pulmonary surfactant apoproteins in lung carcinomas, cancerous tissues from 47 cases were immunohistochemically examined by the avidin-biotin peroxidase complex method using monoclonal antibodies. These antibodies recognized apoproteins in type II pneumocytes in normal and in hyperplastic alveoli and nonciliated columnal cells (Clara cells) in terminal bronchioles of noncancerous tissues. Cancer cells positive for surfactant apoproteins were observed in 11 out of 33 adenocarcinomas, including 1 case of bronchioloalveolar carcinoma. Interestingly, two cases of adenocarcinomas with ultrastructural features of Clara cells, which had metastatic lesions, were positively stained both in the primary and the metastatic sites. Two out of 4 adenosquamous carcinomas showed positive reaction, which was exclusively localized in the cells with papillary structure. The present study indicates that the expression of surfactant apoproteins in cancer cells occurs ubiquitously in adenocarcinomas, which are considered to be derived from the terminal air way.     

Clinicopathologic and immunohistochemical characteristics of goblet cell type adenocarcinoma of the lung.

Twenty-two resected goblet cell type adenocarcinomas of the lung were examined clinicopathologically and immunohistochemically. The stage and survival curve of goblet cell type adenocarcinomas were compared with those of 44 cases of pure or mixed Clara cell and bronchial surface epithelial cell (Clara and BSE) type adenocarcinomas. Each case of goblet cell type was matched with two cases of Clara and BSE type as to sex, age and date of surgery. In goblet cell type adenocarcinomas, lymph node metastasis was less frequently and intrapulmonary metastasis was more frequently detected than in other types of adenocarcinomas (p less than 0.001 and p less than 0.05, respectively). Goblet cell type adenocarcinomas showed better prognoses than Clara and BSE type adenocarcinomas. However, the estimated survival curves of those two groups become similar after adjustment of the TNM condition using Cox's proportional-hazard general linear model. This result indicated that the longer survival of goblet cell type adenocarcinoma was due to the characteristic distribution of TNM conditions, that is, unique local growth and low incidence of lymph node metastasis. When goblet cell type adenocarcinoma was macroscopically classified into two types, i.e. solitary peripheral nodule type (nodular type) and multifocal nodular type or consolidation of all or part of a lobe (diffuse type), the nodular type had better prognosis than the diffuse type (p less than 0.05). Immunohistochemically, 83%, 11%, and 0% of goblet cell type adenocarcinomas were positive for NCC-CO-450, carcinoembryonic antigen (CEA), and surfactant apoprotein, respectively. Most Clara and BSE type adenocarcinomas were negative for NCC-CO-450, but positive for CEA and surfactant apoprotein.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunoreactivity of surfactant-apoprotein in adenocarcinomas, large cell and small cell carcinomas of the lung

Fifty seven adenocarcinomas, 43 large cell and 30 small cell lung carcinomas were immunohistochemically examined using monospecific IgG against pulmonary surfactant apoprotein. Six peripheral adenocarcinomas and one peripheral large cell carcinoma were found to be histogenetically related to type II pneumocytes. They showed an acinar, papillary or solid growth pattern. The immunohistochemical study using anti-surfactant apoprotein IgG gave a granular reaction product in both the cytoplasm and nucleus of tumor cells whose intensity was variable. Intranuclear inclusions were generally the most densely stained structures. These results suggest that lung carcinomas derived from alveolar type II cells are found not only in bronchioloalveolar tumors, but are also found in other types of adenocarcinoma and in large cell carcinomas.     

Extremely high Langerhans cell infiltration contributes to the favourable prognosis of HPV-infected squamous cell carcinoma and adenocarcinoma of the lung

AIMS: The infiltration of Langerhans cells in adenocarcinomas and squamous cell carcinomas of the lung was examined in relation to prognostic implications and human papillomavirus (HPV) infection. METHODS AND RESULTS: Samples from 62 adenocarcinoma and 59 squamous cell carcinoma patients in 1995-97, the prognosis of which had been followed up, were used. The Langerhans cells were demonstrated immunohistochemically using anti S100a and CD1 antibodies. Human papillomavirus (HPV) infection was examined by polymerase chain reaction (PCR) and nonisotopic in-situ hybridization (NISH) methods. Statistical analysis was carried out using the Kaplan-Meier method (Wilcoxon analysis) and multiple regression analysis. HPV infection was demonstrated in 12 cases (19.4%) of adenocarcinoma. The HPV-infected adenocarcinomas had abundant faintly eosinophilic cytoplasm, and were immunohistochemically positive for the surfactant apoprotein A. In the 59 cases of squamous cell carcinomas 19 were of the well differentiated form, and 29 and 11 were moderately and poorly differentiated cases, respectively. HPV was detected in 29 cases (49.2%) (13 well and 16 moderately differentiated cases). In all HPV-infected adenocarcinoma and squamous cell carcinoma cases, extremely large numbers of Langerhans cells (more than 100 per high-power field) were demonstrated in the tumour nests. In contrast, in the non-HPV-infected adenocarcinomas and squamous cell carcinomas, only a few (less than about 10 per high-power field) Langerhans cells were observed. The squamous cell carcinoma cases with high Langerhans cell infiltration, which were also infected with HPV, showed a significantly good prognosis (P = 0.007). The adenocarcinoma cases with high Langerhans cell infiltration tended to have a better prognosis than the cases with low Langerhans cell infiltration, but the difference was not statistically significant. The low number of highly infiltrated cases was insufficient for an adequate statistical analysis. Furthermore, there was no significant correlation between either Langerhans cell infiltration and smoking, or HPV infection and smoking, in either squamous cell carcinoma or adenocarcinoma cases. CONCLUSIONS: It was considered that the extremely high Langerhans cell infiltration in the tumours was caused by HPV infection. The extremely large number of Langerhans cells in the tumours contributes to the favourable prognosis for HPV-infected lung cancer.     

Histochemical characterization of non-neuroendocrine tumors and neuroendocrine cell hyperplasia induced in hamster lung by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone with or without hyperoxia.

Lung tumors induced by 4-(methylnitrosamine)-1-(3-pyridyl)-1-butanone (NNK) with or without hyperoxia have frequent K-ras mutations but only rare p53 mutations, suggesting that this may be a model for non-small cell lung cancers. The goals of the present study were (1) to characterize the histopathology of lung tumors induced in hamsters by NNK with or without O2 and (2) as a corollary, to quantitate the pulmonary neuroendocrine cell hyperplasia in the different treatment groups early and late in the treatment period. Lung tumors induced by NNK with or without O2 were 71% adenomas, 22% adenocarcinomas, approximately 4% bronchoalveolar carcinomas, and approximately 4% squamous/adenosquamous carcinomas. One-half of all tumors were positive for the Clara cell antigen CC10 and 21% of NNK-induced tumors were mucin positive, compared with 2% of NNK/O2-induced tumors (P = 0.003). Immunostaining for PGP9.5 was positive in 5% of tumors induced by NNK alone, but in none of NNK/O2-induced tumors (P = 0.024). Abundant proliferating cell nuclear antigen occurred in 55% of NNK-induced tumors, compared with 19% of NNK/O2-induced tumors (P = 0.009). These data indicate that NNK with or without O2 induces non-neuroendocrine lung tumors. Hyperoxia appears to inhibit cell proliferation and suppress mucinous and partial neuroendocrine differentiation in some of these tumors.     

Expression of alpha-methylacyl-CoA racemase (P504s) in various malignant neoplasms and normal tissues: astudy of 761 cases

Alpha-methylacyl CoA racemase (AMACR), also known as P504S, plays an important role in peroxisomal beta-oxidation of branched-chain fatty acids. It has recently been shown that AMACR is highly expressed in prostate cancer and that it may be an important diagnostic marker for prostate carcinoma. However, little is known about expression of AMACR in normal tissues and other malignant tumors. In this study, we investigated expression of AMACR in 539 malignant tumors and 222 normal human tissues of various types by immunohistochemical analysis. mRNA levels of AMACR in normal organs and in selected tumors were assessed by real time PCR. In normal tissue, high expression of AMACR mRNA was identified in liver, kidney and salivary gland, while AMACR protein was detected in liver (hepatocytes), kidney (tubular epithelial cells), lung (only bronchial epithelial cells), and gallbladder (only mucosal epithelial cells). High expression of AMACR mRNA was found in prostate, liver, and kidney cancers but rarely in stomach and bladder cancers. A high percent of adenocarcinomas arising from these organs express AMACR, including 17 of 21 (81%) of hepatocellular carcinomas and 18 of 24 (75%) of renal cell carcinomas. In addition, carcinomas arising from tissues normally not expressing AMACR were also positive for the antigen, including 17 of 18 (94%) prostate carcinomas, 9 of 29 (31%) of urothelial carcinomas, and 4 of 15 (27%) of gastric adenocarcinomas. Two hundred and fifty cases of adenocarcinomas from lung, breast, pancreas, bile duct, adrenal gland, salivary gland, ovary, thyroid and endometrium were negative or rarely positive for AMACR. Neuroendocrine carcinomas rarely expressed AMACR. Melanomas, squamous cell carcinomas, basal cell carcinomas, soft tissue tumors (including epithelioid sarcomas and synovial sarcoma), thymomas, and germ cell tumors were negative for AMACR. Our data provide important baseline information for using AMACR in clinical practice and also are valuable in furthering understanding of the pathogenic role of AMACR in malignant neoplasms.     

Lowp53 protein expression in salivary gland tumours compared with lung carcinomas

Summary Fifty-one salivary gland tumours (23 pleomorphic adenomas, 5 Warthin's tumours, 12 mucoepidermoid carcinomas, 7 adenoid cystic carcinomas, 3 undifferentiated carcinomas and 1 acinic cell tumour) and 27 lung carcinomas (18 squamous cell carcinomas, 6 adenocarcinomas and 3 small cell carcinomas) were analysed immunohistochemically for the expression ofp53 nuclear phosphoprotein. Eight out of 51 (16%) salivary gland tumours werep53 positive. Three of these were benign and 5 malignant. All 3 benign salivary gland tumours were pleomorphic adenomas and expressed only occasional nuclear positivity with less than 1% of tumour cells positive. Of the 5p53-positive malignant tumours, 3 were mucoepidermoid carcinomas and 2 undifferentiated carcinomas. The malignant salivary gland tumours expressed more than 1% of positive nuclei in every case. Seventeen lung carcinomas werep53 positive (63%). Thirteen of these were squamous cell carcinomas, 3 were adenocarcinomas and 1 small cell lung carcinoma. The results show that mutations of thep53 gene may be infrequent in salivary gland tumours when compared with lung carcinomas. The relatively indolent course of some histological types of malignant salivary gland tumours could be associated with the preservation of the non-mutatedp53 gene in most of these tumours. The presence ofp53 positivity in some pleomorphic adenomas might, on one hand, suggest thatp53 gene alterations are also present in these tumours; on the other hand, the accumulation of thep53 protein in these tumours might also be due to some unknown mechanism, not necessarily related top53 gene mutation.     

IMMUNOREACTIVITY OF SURFACTANT-APOPROTEIN IN ADENOCARCINOMAS, LARGE CELL and SMALL CELL CARCINOMAS OF THE LUNG

Fifty seven adenocarcinomas, 43 large cell and 30 small cell lung carcinomas were immunohistochemically examined using monospecific IgG against pulmonary surfactant apoprotein. Six peripheral adenocarcinomas and one peripheral large cell carcinoma were found to be histogenetically related to type II pneumocytes. They showed an acinar, papillary or solid growth pattern. The immunohistochemical study using anti-surfactant apoprotein IgG gave a granular reaction product in both the cytoplasm and nucleus of tumor cells whose intensity was variable. Intranuclaer inclusions were generally the most densely stained structures. These results suggest that lung carcinomas derived from alveolar type II cells are found not only in bronchioloalveolar tumors, but are also found in other types of adenocarcinoma and in large cell carcinomas. ACTA PATHOL. JPN. 36: 1271 ∼ 1278, 1986.     

Restoration of the normal Clara cell phenotype after chronic allergic inflammation

Bronchiolar Clara cells play a critical role in lung homoeostasis. The main goal of this study was to evaluate the effects of chronic allergy on these cells and the efficacy of budesonide (BUD) and montelukast (MK) in restoring their typical phenotypes after ovalbumin‐induced chronic allergy in mice. Chronic allergy induced extensive bronchiolar Alcian blue‐periodic acid‐Schiff (AB/PAS)‐positive metaplasia. In addition, cells accumulated numerous big electron‐lucent granules negative for Clara cell main secretory protein (CC16), and consequently, CC16 was significantly reduced in bronchoalveolar lavage. A concomitant reduction in SP‐D and CYP2E1 content was observed. The phenotypic changes induced by allergy were pharmacologically reversed by both treatments; MK was more efficient than BUD in doing so. MK decreased AB/PAS reactivity to control levels whereas they remained persistently elevated after BUD. Moreover, most non‐ciliated cells recovered their normal morphology after MK, whereas for BUD normal cells coexisted with ‘transitional’ cells that contained remnant mucous granules and stained strongly for CC16 and SP‐D. Glucocorticoids were also less able to reduce inflammatory infiltration and maintained higher percentage of neutrophils, which may have contributed to prolonged mucin expression. These results show that chronic allergy‐induced mucous metaplasia of Clara cells affects their defensive mechanisms. However, anti‐inflammatory treatments were able to re‐establish the normal phenotype of Clara cell, with MK being more efficient at restoring a normal profile than BUD. This study highlights the role of epithelial cells in lung injuries and their contribution to anti‐inflammatory therapies.     

Histochemical reactivities of lectins and surfactant apoprotein in pulmonary adenocarcinomas and their metastases.

In an attempt to better understand the biologic behavior of neoplastic cell metastasis, a histochemical study with the use of six different lectins and a monoclonal antibody against human pulmonary surfactant apoprotein (PE-10) was carried out on primary adenocarcinomas of the lungs and their regional (usually lymphatic to lymph nodes or contralateral lung) and distant (usually hematogenous to extrathoracic organs) metastatic lesions of 54 postmortem cases. Primary pulmonary adenocarcinomas were classified further into acinar, papillary, and solid types according to WHO histological typing. Acinar type primary adenocarcinoma of the lungs showed significantly higher (p less than 0.05) binding reactions to Ricinus communis-I (RCA-I) and Ulex europaeus-I (UEA-I) lectins than solid type adenocarcinoma. With six different lectins, concordantly positive reactions between primary pulmonary adenocarcinomas and their lymphatic and hematogenous metastases were seen in 67% or more cases, and with soybean agglutinin (SBA) and UEA-I the concordance rates between primary and lymphatic metastases (lymph nodes and contralateral lungs, respectively) were significantly higher (p less than 0.05) than those between primary and hematogenous metastases. With PE-10 immunohistochemistry, concordantly positive reactions between primary and metastases were low, especially in cases of distant hematogenous metastases (25%), but the statistical significance of differences was missed by narrow margins. With alcian blue PAS-stain, concordantly positive reactions of mucin production between primary adenocarcinomas and both lymphatic and hematogenous metastases were high (92%), but there was no evidence of correlation between lectin bindings and alcian blue-PAS reactions to either primary or metastatic lesions of pulmonary adenocarcinomas.     

Decrease of the surface fraction of surfactant proteins containing clara cells and type II pneumocytes in a rat asthma model.

In asthma surfactant proteins (SP) might differ in distribution and composition and thus play a role in pathophysiology of this disease. Therefore, the well-established animal model of ovalbumin sensitized and challenged rats were used to study the distribution of surfactant proteins in Clara cells and type II pneumocytes. Serial sections of paraffin embedded lung tissue were sequentially immunostained by the avidin-biotin-peroxidase complex (ABC) technique. Antisera against SP-A, SP-B and Clara cell specific protein (CC10) were used. We determined stereologically' the surface fraction of immunolabelled cells and semiquantitatively the percentage of test fields containing labelled alveolar macrophages. In allergen sensitized and provocated rat lungs: (1) the surface fraction of SP-A and SP-B positive Clara cells was significantly reduced, (2) the surface fraction of Clara cells stained with CC10 was coincided with controls, (3) the surface fraction of SP-A and not of SP-B possitive type II pneumocytes decreased significantly, (4) a significantly higher percentage of test fields with SP-A labelled alveolar macrophages was evaluated. Thus, in this animal model of asthma the inflammatory process after allergen challenge is accompanied by alterations in the distribution patterns of SP in Clara cells and type II pneumocytes.