Characterization of the spreading process in human T lymphocytes

Substratum-bound concanavalin A (conA) caused attachment and spreading of human T lymphocytes identified by monoclonal anti-T cell antibodies and sheep erythrocyte rosette formation. The simultaneous presence of conA in the medium increased the spreading, whereas preincubation of the cells with conA inhibited spreading. The tendency of conA to induce spreading was dependent on the concentration used, the higher the conA concentration the more pronounced was the spreading. For example, conA at 10 micrograms ml-1 triggered the formation of prominent substratum-attached filopodia with a length of 1-10 micron in 60-80% of T-enriched lymphocytes obtained from separate individuals. At the same conA dose the filopodia were, in 10-20% of the lymphocytes, accompanied by development of lamellipodia. With conA at 100 micrograms ml-1 the number of cells that underwent pronounced spreading was 55-90% in separate individuals. Observation of T-enriched cells fixed at different times after initiation of spreading induced by conA at 100 micrograms ml-1 indicated that filopodia formation represented the initial morphological alteration during the spreading process. This process thereafter proceeded with development of lamellipodia, extensive cytoplasmic spreading and flattening of the central cell mass. Quiescent and mitogen-activated cells exhibited the same sequence of changes during spreading. Spreading led to disappearance of the microvilli with a length of 0.1-0.7 micron present on lymphocytes in suspension, although some microvilli persisted over the cell center.