Immunohistochemical detection of human epidermal growth factor in submandibular glands and their tumors using a polyclonal antiserum and a monoclonal antibody

Abstract

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, spindle-shaped tumor cells and chondroidally changed tumor cells also exhibited positive staining for hEGF.

An Ontogenetic Study on Endothelin-Producing Cells in the Rat Lung and Submandibular Gland.

“…The secretory granules of GCT cells, which are more prominent in male rodents than in females, have been shown to contain nerve growth factor (2,4,22), epidermal growth factor (2,5,22), renin (2,9), and esteropeptidases (2,7). Epidermal growth factor has also been reported to exist in the normal ductal system and ductal origin tumor cells of humans (28), in addition to atrial natriuretic polypeptide in the submandibular glands of rats (21). ET, which may have cellproliferation and -differentiation activity, is thought to co-exist in these cells with the above-mentioned bioactive substances and to be co-secreted from GCT cells into saliva.…”

Widespread distribution of human epidermal growth factor(hEGF) in human tissues and organs under normal and tumorous conditions. Immunohistochemical studies with the use of polyclonal and monoclonal antibodies.

“…When monoclonal antibody was employed, the indirect PAP method was done. The details of the methods were reported in a previous paper (53). In the control method, representative sections from normal tissues and organs were tested.…”

“…For generation of antiserum, both highly purified hEGF from human urine (45) and hEGF produced biotechnologically by the synthetic gene method (39,55) have been used as immunogens. A preceding paper has described the details of the method for immunohistochemical detection of hEGF and the localization of hEGF in normal salivary glands as well as in their neoplastic lesions (36,53). The present study describes the widespread distribution of immunohistochemically detectable hEGF in human tisr:ies and organs under normal and neoplastic conditions.…”

Immunohistochemical localization of human epidermal growth factor/.GAMMA.-urogastrone in submandibular glands and in their obstructive lesions.

“…Those cells might corresponding to centroacinar cells of pancreatic acini. Positive hEGF staining has also been reported in salivary gland ducts with the use of monoclonal antibody with the production of synthetic gene method (26). In mouse SMG, mEGF is positive in GCT cells which differentiate from ducts during postnatal development (17).…”

“…It has been termed hEGF since primary structure and biological activities resemble those of mEGF (4), and it is now accepted that hEFG and UG are the same (5,8). Immunohistochemical localization of hEFG in tissues and organs has already been reported by use with anti-hEGF antiserum obtained from human urine (6,11,14,19), and monoclonal antibody against hEGF or polyclonal hEGF antiserum generated in response to synthetic gene technique with the use of E. coli (26). Immunohistochemical expression of hEGF/UG in human salivary glands has been controversial; Elder et al (6) stated that hEGF/UG was present in ductal cells, while Heitz et al (11) and Kasselberg et al .…”