Destruction of Epidermal Cells in Vitro by Autologous Serum From Normal Animals

Abstract

Although normal serum is sometimes said to be toxic to autologous cells in vitro (1), to our knowledge, data of such autotoxic effects have been previously reported only for spermatozoa (2, 3). Indeed, autologous serum is often used as the best nutrient medium for cells (4). Evidence will be presented here that fresh normal sera from rabbits and rats are lethal in vitro to autologous epidermal cells. In the test system employed, 70 to 100 per cent of epidermal cells were killed within 2 hours at 37°C. Materials and MethodsAnimals.--The following strains were used: Dutch and Zealand rabbits, Long Evans rats, C57 mice, and mongrel guinea pigs.Cell Preparation.--Epidermal cells from the basal or malpighian layer of the epidermis were prepared by the trypsinization method of Billingham and Reynolds (5) with a few modifications. A 0.010 to 0.015 inch split thickness of skin was obtained from the lateral surface of the body with a Brown electrodermatome. It was then scraped free of dead cuticular cells and connective tissue, and placed with its dermal surface down in a Petri dish containing 0.25 per cent trypsin (Difco 1:250) in versene-BSS (0.002 ~ tetrasodium ethylenediaminetetraacetate in Hanks's balanced salt solution). Upon incubation in a shaker at 37°C for 20 minutes, areas of the skin which appeared clear were excised and the remaining skin allowed to incubate for periods up to 1~ hours. The trypsinized pieces were then washed in saline and in versene-BSS, placed with the dermis of the skin up in a dry Petri dish, and blotted free of fluid. After peeling back the dermis the exposed surface of the epidermis was scraped with a scalpel. The freed malpighian cells were suspended in versene-BSS, pooled, centrifuged at 1000 g for 1 minute, and resuspended in BSS. Later tests have indicated that the use of versene could be eliminated completely.

Heterophile Antigen Detected by Infectious Mononucleosis Sera on Lymphoid Cells

“…Both NHS and IMS produced lysis of murine thymocytes. This ob servation was in agreement with studies by other investigators, which demonstrated the cytotoxicity of heterologous sera for murine thymo cytes [9][10][11][12]. Significantly higher titers of IMS than of NHS suggested that lysis of thymocytes could be caused by heterophile antibodies pre sent in IMS in addition to naturally occurring thymocytolysins.…”

Specificity of Lymphocytotoxic Anti‐Human Antibodies in Normal Rabbit Sera

“…The loss of activity of normal rabbit sera after storage confirms previous observations (Terasaki et al 1961) that normal sera cytotoxic to lymphocytes from other species often become inactive after stor-age. Tests with five sera indicated that the loss of activity due to storage at -20" C was not due to anticomplementarity of the sera.…”

“…Discussion 'That normal rabbit sera contain cytotoxic antibodies directed against human lymphoid cells has been known for several years (Terasaki et al 1961, Herberman 1969, Ferrone et al 1971. Although these antibodies detect antigens on cell surfaces of lymphocytes of normal individuals, they do not appear specific for known H G A antigens.…”

A Study of Auto-Allergy in Generalized Eczema.

“…In a very different approach Terasaki and Chamberlain (1962) found that autologous serum from normal rabbits and rats could be toxic in vitro when incubated with their own epidermal cells, though it remains to be determined wivat signiticanco this may have in auto-allergy.…”