Antigenic Differences between Normal Mouse Epidermis and Methylcholanthrene-Induced Squamous-Cell Carcinomas

Abstract

IgG preparations were made from antisera raised against the urea- and alkali-extracted antigens of normal mouse epidermis and methyl- cholanthrene-induced squamous-cell carcinomas. Immunodiffusion in agar with thoroughly adsorbed IgG immuno- globulins showed that the urea- and alkali-extracted proteins of the carcinomas had antigens in common. Antigens in common were also found in the epidermal urea-and alkali-extracted proteins, but the antigens common to the carcinoma proteins were not identical with the antigens common to the epidermal proteins. Preparations of antibodies against epidermal and carcinoma urea-extracted proteins showed specific adsorption to epidermal and carcinoma sediments, respectively. There was no significant in vivo localization of ¹²⁵I-labeled antibodies against epidermal and carcinoma urea-extracted protein in spleen, kidney, liver, and epidermis in normal mice, or in spleen, kidney, liver, and carcinomas in carcinoma-bearing mice.

Differences in the Urea-extracted Proteins of Mouse Epidermis and Squamous cell Carcinomata Determined by Fluorescence Microscopy

“…The studies reported here on the urea proteins of epidermis, papilloma and carcinoma by fluorescence microscopy confirm the antigenic differences observed by immunodiffusion (Carruthers, 1970), and with the paired label technique (Bhattacharaya and Carruthers, 1972). The ureaextracted proteins of mammalian epidermis consist of several species as indicated by molecular sieve chromatography, and these components have a common antigenic determinant (Carruthers and Bhattacharaya, 1972a;Tezuku and Freedberg, 1972).…”

“…The investigations of Rudall (1952) indicate that the fibrous or structural (urea-extracted) proteins of epidermis arise from the tonofilaments and are thus intracellularly derived. In this connection radiolabelled antibodies prepared against epidermal urea proteins did not penetrate epidermal cells in vivo (Bhattacharaya and Carruthers, 1972). Actually, Tezuka et al (1972) (Fig.…”

“…These are: (a) preparation of normal and hyperplastic mouse epidermis, methyleholanthrene-induced papilloma and carcinoma; (b) isolation of the urea-extractable proteins of mouse epidermis, papilloma and carcinoma; (c) preparation of antisera raised in rabbits against these ureaextractable proteins; and (d) preparation of mouse tissue sediments for the absorption of antisera (Carruthers, 1970;Bhattacharaya and Carruthers, 1972). The titres of the antisera were determined by the radial immunodiffusion procedure of Masseyeff and Zisswiller (1969), and by the determination of the dilution of serum and tissue-absorbed antisera Awhich still stained the tissues by fluorescence microscopy.…”

“…ANTIGENIC differences in the ureaextractable proteins of normal and hyperplastic mouse epidermis, papilloma and squamous cell carcinoma have been demonstrated Baumler, 1965, 1966;Carruthers and Bhattacharaya, 1 972b;Carruthers, 1970;Bhattacharaya and Carruthers, 1972). These ureaextracted proteins are extracted with 6 mol/l urea and precipitated maximally at pH (Carruthers et al, 1955(Carruthers et al, , 1957Rudall, 1.952), and are sometimes denoted as the urea-extractable 5-5 proteins; herein for simplicity they are called the urea proteins.…”

“…Previous studies indicated that epidermis contained urea-extractable antigens which were absent, or present in small amounts, in carcinoma and that carcinoma contained urea-extractable antigens which were absent or present in small amounts in epidermis. These differences were demonstrated by using thoroughly absorbed antisera or immunoglobulins (raised in rabbits against the urea proteins of epidermis, papilloma and carcinoma) by (1) fluorescence microscopy; (2) immuunodiffusion in agar; and (3) the paired label technique using 131 and 1 2 51 (Carruthers and Baumler, 1966;Carruthers, 1970;Bhattacharaya and Carruthers, 1972).…”