Summary
We applied immunohistochemical procedures to detect hEGF in salivary glands and pleomorphic adenomas of salivary-gland origin using a polyclonal hEGF antiserum and a monoclonal antibody against hEGF synthesized by applying the synthetic gene technique using Escherichia coli. In normal salivary glands, hEGF was mainly localized in the ductal system (i.e., intercalated, striated, and excretory ducts). The staining intensity and intracellular localization exhibited some variation depending on the fixative used. When a polyclonal hEGF antiserum was used for immunostaining, slight background staining was observed in sections prepared using the fixatives tested. Therefore, precise localization of hEGF was obtainable only in formalin-fixed sections using the monoclonal antibody against hEGF. In pleomorphic adenomas, positive hEGF staining was seen on the luminal side of tumors and in cells of ductal origin; no reactivity was present on the outer side of tumors or in cells of myoepithelial origin. Occasionally, long, spindleshaped tumor cells and chondroidally changed tumor cells also exhibited positive staining for hEGF.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cohen S, Carpenter G (1975) Human epidermal growth factor; isolation and chemical and biological properties. Proc Natl Acad Sci USA 72:1317–1321
Dailey GE, Kraus JW, Orth DN (1978) Homologous radioimmunoassay for human epidermal growth factor (Urogastrone). J Clin Endocrinol Metab 46:929–939
Elder JB, Williams G, Lacey E, Gregory H (1978) Cellular localisation of human urogastron/epidermal growth factor. Nature 271:466–467
Gregory H (1975) Isolation and structure of urogastrone and its relationship to epidermal growth factor. Nature 257:325–327
Hayashi K, Namoto H, Kurobe M, Nishimura S, Hiratani H, Furukawa S (1985) A sensitive two-site enzyme immunoassay for human epidermal growth factor (urogastrone). Biochem Inter 10:839–845
Heitz PU, Kasper M, Van Noorden S, Polak JM, Gregory H, Pearse AGE (1978) Immunohistochemical localisation of urogastrone to human duodenal and submandibular glands. Gut 19:408–413
Hirata Y, Orth DN (1979a) Epidermal growth factor (urogastrone) in human tissue. J Clin Endocrinol Metab 48:667–672
Hirata Y, Orth DN (1979b) Epidermal growth factor (urogastrone) in human fluids: size heterogene. J Clin Endocrinol Metab 48:673–679
Kasselberg AG, Orth DN, Gray ME, Stahlman MT (1985) Immunocytochemical localization of human epidermal growth factor/urogastrone in several human tissues. J Histochem Cytochem 33:315–322
Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificoty. Nature 256:495–497
Kurobe M, Furukawa S, Hayashi K (1985) Synthesis and secretion of an epidermal growth factor (EGF) by human fibroblast cells in culture. Biochem Biophys Res Commun 131:1080–1085
Mori M, Hamada K, Naito R, Tsukitani K, Asano K (1983) Immunohistochemical localization of epidermal growth factor in rodent submandibular glands. Acta Histochem Cytochem 16:536–548
Mori M, Murase N, Hyun KH, Sumitomo S, Kawamura K (1985) Immunohistochemical studies of keratin distribution in salivary gland tumors. Acta Histochem Cytochem 18:21–32
Mori M, Sumitomo S, Iwai Y, Meenagham MA (1986a) Immunolocalization of keratins in salivary gland pleomorphic adenoma using monoclonal antibodies. Oral Surg 61:611–616
Mori M, Murase N, Hosaka M, Orito T (1986b) Immunohistochemical expression of S-100 protein in reactive and neoplastic myoepithelial cells of variant salivary pleomorphic adenomas. Acta Histochem Cytochem 19:231–240
Oka T, Sakamoto S, Miyoshi K, Fuwa T, Yoda K, Yamasaki M, Tamura G, Miyake T (1985) Synthesis and secretion of human epidermal growth factor by Escherichia coli. Proc Natl Acad Sci USA 82:7212–7216
Poulson SS, Nexo E, SkovOlson P, Hess J, Kirkegaard P (1986) Immunohistochemical localization of epidermal growth factor in rat and man. Histochemistry 85:389–394
Sato M, Yoshida H, Hayashi Y, Miyakami K, Bando T, Yanagawa T, Yura Y, Azuma M, Ueno A (1985) Expression of epidermal growth factor and transforming factor-β in a human salivary gland adenocarcinoma cell line. Cancer Res 45:6160–6167
Savage CR, Harper R (1981) Human epidermal growth factor/urogastrone rapid purification procedure and partial characterization. Anal Biochem 111:195–202
Starkey R, Orth DN (1977) Radioimmunoassay of human growth factor (urogastrone). J Clin Endocrinol Metab 45:1144–1153
Starkey RH, Cohen S, Orth DN (1975) Epidermal growth factor. Identification of a new hormon in human urine. Science 189:800–802
Takai Y, Sumitomo S, Noda Y, Asano K, Mori M (1985) Immunohistochemical observation of EGF and NGF in submandibular glands after duct ligation with or without testosterone administration. J Oral Pathol 14:322–331
Tahara E, Sumiyoshi H, Hata J, Yasui W, Taniyama K, Hayashi T, Nagae S, Sakamoto S (1986) Human epidermal growth factor in gastric carcinoma as a marker of high malignancy. Jpn J Cancer Res 77:145–152
Tsukitani K, Tatemoto Y, Noda Y, Mori M, Kurobe M, Hayashi K (1987) Immunohistochemical localization of human EGF/urogastrone in submandibular glands and in their obstructive lesions. Acta Histochem Cytochem (in press)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Tsukitani, K., Tatemoto, Y., Noda, Y. et al. Immunohistochemical detection of human epidermal growth factor in submandibular glands and their tumors using a polyclonal antiserum and a monoclonal antibody. Histochemistry 87, 293–300 (1987). https://doi.org/10.1007/BF00492581
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00492581