A and B Antigens in the Human Epidermis 1

Abstract

Several attempts to demonstrate the presence of A and B antigens in the human epidermis have been undertaken in the past, but with negative results (1, 2). Recently, Coombs, Bedford, and Rouillard (3) proved by the mixed agglutination method the existence of A and B group specific substances in epidermal cells.In continuation of studies on the A and B antigens in the human cornea (4, 5) the presence of these antigens in the human epidermis was investigated.The following is a report of this investigation. MATERIAL AND METHODSThin split thickness skin was obtained by means of an electric dermatome from 19 patients, selected at random, who were admitted to the hospital for various skin grafting procedures. No differentiation was made between secretors and non-secretors. Part of this skin was used for skin grafting, the rest rinsed in physiological saline and despatched to the laboratory, completely immersed in a container full of saline. There the excised skin was cut to standard size, 3 X 3 cm. and washed 3 times in 200 ml. normal saline. The flask containing saline and skin was agitated vigorously on a Kahn shaking machine for 10 minutes. In order to test for complete removal of erythrocytes, the last saline washing was centrifuged in large conical tubes and the minute sediments examined microscopically for red blood cells. If only one erythrocyte was found, three additional washings were carried out. Every sediment was further tested by the Benzidine Reaction (6). This was sometimes found to be positive after the first washing, but always negative after the third. After this treatment the skin was used either fresh or frozen at -200 C in 2 ml. of normal saline. For every experiment 0.5 X 0.5 cm. pieces of skin were cut into very thin slices with the aid of fine scissors and scalpels. These slices were homogenized for 30 minutes in a small pyrex glass tissue grinder with a pyrex glass pestle connected to a variable speed stirring apparatus. Five-tenths ml. of saline was added to the homogenate during homogenization. To prevent excessive heat production the grinder was immersed in 1 This study was aided by a grant from the Hematology Research Foundation, Chicago.an ice bath and the stirring carried out at intervals. The fine white pasty mass obtained served as antigenic material. Preparation of sera. Sera were inactivated at 560 C for 30 minutes, and titrated for anti-A and anti-B isoagglutinins. Only sera with the minimal titer of 1:128 when undiluted were used for absorption experiments. Sera were distributed in small quantities stored at -20°C until used; no serum was frozen and thawed more than once.Isoagglutinin determination. Titrations of isoagglutinins were performed by the double serial dilution method against freshly prepared 2 per cent erythrocyte suspensions. Results were read after allowing the tubes to stand at room temperature for 30 minutes. The titer was expressed as the last dilution at which agglutinates could be detected by the naked eye. The tubes were neither centrifuged nor examined micros...

The Histological Distribution of Blood Group Substances a and B in Man

“…The staining of the latter, it will be recalled, is abolished by prior exposure to alcohol. Localization of antigen in the Malpighian (and granular) layers of stratified squamous epithelia provides histologic confirmation of absorption of anti-A and anti-B agglutinins by skin homogenates (21) and of the mixed agglutination of suspended epidermal cells (2).…”

“…More recently there has been a revival of interest in cell and tissue antigens with the reapplication of old and the invention of new techniques. A and B antigens have been found in the cornea (22) and the skin (21) by the isoagglutinin absorption method. Coombs used the technique of mixed cell agglutination to demonstrate A and B antigens in epidermis (2) and in platelets (1).…”

The Destruction of Red Cells by Antibodies in Man. II. Pyrogenic, Leukocytic and Dermal Responses to Immune Hemolysis1

“…Davidsohn and his associates (63) have described suppurative skin lesions in rabbits at the site of injection of anti-rabbit-red cell hemolytic immune serum. The authors postulated that this effect reflected an antigenic similarity between red cells and skin; and, indeed, it is known that human epidermis may contain A and B antigens (64,65). However, the skin reactions described here were minimal after direct injections of antiserum, and required for full development the presence of red cells.…”

PBS-soluble substances with blood group activity from human epidermis and dermis

“…as de termined by IF technique. Homogenates of whole skin were found to absorb antibodies against blood group antigens A and B ( 13).…”