Surface mapping of mouse thymocytes

Abstract

The blocking method used previously for determining the relative positions of different components of the cell surface was modified by first fixing the cells with paraformaldehyde. This technique was applied to the H-2K (K), H-2D (D), TL, Lyt-1, and Lyt-2 surface components of mouse thymocytes, and the results were compared in parallel with data obtained with the original technique with unfixed cells. Previous mapping data with unfixed cells, indicating the positions of these molecules relative to one another, were confirmed with paraformaldehyde-fixed cells, with one exception. On unfixed cells, D and TL appeared sufficiently adjacent to produce mutual interference in the attachment of anti-D and anti-TL antibodies. With paraformaldehyde-fixed cells this was not so, D and TL appearing sufficiently separated from one another to obviate interference in the attachment of anti-D and anti-TL antibodies. The previously reported close association of K with Lyt-1 and of D with Lyt-2 were demonstrable equally with unfixed and paraformaldehyde-fixed thymocytes. It is suggested that activation of D sites, and alternatively of TL sites, by antibody in the present experiments brings these two molecules into apposition and that this movement may exemplify a mechanism concerned in immunological recognition and response.