Redistribution of surface antigens--a general property of animal cells?

Abstract

MEMBRANE antigens of animal cells are reported to be localized either in discontinuous discrete areas (H-21,2, HL-A3, organ4 and immunoglobulin (Ig)5–9 or continuously (species4 and blood group9,10). These results have been obtained by immunofluorescence (IFL) and electron microscopy performed by double layer principles except in the case of Ig where direct IFL technique has been used. The influence on the localization pattern of marker molecules by variables such as Ig concentration of the reagents used, effects of interactions between Ig molecules in the double layer and temperature dependence of the reaction steps have not been investigated. Redistribution of lymphocyte surface Ig molecules induced by anti-Ig antibody has been described and this was observed to be temperature dependent occurring at 20° C but not at 0° C11,12. Here I discuss the distribution and localization of different surface antigens (blood group substance A and B, species, H-2 and organ) on several cell types (monkey kidney, human lymphoid cells, HeLa, human thyroid cells, RK13 and L-cells of C3H/K) as recorded by direct and indirect IFL technique. I show that the temperature and Ig concentration used exert an influence on the distribution of these antigens. The cells used in IFL tests were cultured and harvested as described before10. IFL staining was performed according to Moller13. The stained cells were examined with a Leitz ‘Orthoplan’ ultraviolet microscope. Direct IFL technique was applied to two test systems. A FITC-conjugated rabbit antiserum to blood group substance A was used on monkey (Macaca fascicularis) kidney cells known to be A-positive10 and a FITC-conjugated horse anti-human thymocyte globulin were tested on a human lymphoid cell line Robinson (obtained from Dr Moore, Buffalo). In both test systems the staining pattern was completely confluent at all temperatures used: 4° C, 20° C and 37° C. The staining pattern was not related to antibody concentration.