[Influence of several experimental conditions on the quantity of E- and EAC-rosette forming cells in peripheral human...

The authors analysed the influence of some conditions of studying the rosette-formation between the human peripheral blood lymphocytes and sheep red blood cells, not loaded (a) and loaded with antibodies and a complement (b). It was shown that rosette formation depended on the temperature at which the experiment was conducted. Thus, after the incubation of a mixture of cells at 37 degrees C, and then at 4 degrees C the number of rosette-forming cells was greater than after the incubation at 4 degrees C alone. Substances used for their fixation also influenced the formation of rosettes: in the treatment of erythrocytes with formalin, tannin and glutaraldehyde, as well as introduction of the latter into the mixture of erythrocytes and leukocytes considerably increased the number of a and b (the sum of T- and B-lymphocytes exceeded 100%). Among a and b in such experimental conditions there was seen a great number of monocytes and of segmented leukocytes. Fixation of smears prepared from a mixture of cells after the incubation in formalin vapour did not lead to such increase in the number of rosette-forming cells. Apparently in case of using glutardehyde, formalin and tannin cells for the treatment of cells additional investigations are necessary with the purpose of identification of a and b as T- and B-lymphocytes.