In a previous study (Nakashima et al., Eur. J. Immunol., 20: 47-53, 1990), a cloned stromal cell line TEL-2 was established from Balb/c mouse thymus. Incubation of thymocytes with TEL-2 cells resulted in the selective elimination of CD4 and CD8 double-positive thymocytes from the culture. In the present report, both phase-contrast and scanning electron microscopes were used to examine, at various time intervals, TEL-2 cells cocultivated with thymocytes in order to elucidate the kinetic sequence of their cellular interaction. The thymocytes attached to the TEL-2 cell surface were more numerous at early times (30 min to 1 h), and their number decreased gradually with time. In contrast, the thymocytes that migrated into the TEL-2 cell layers were less abundant at early times, their number increasing with time thereafter. Destruction of the regular arrangement of TEL-2 cells was found at later than 1 h, suggesting active cellular interaction. The thymocytes adherent to the TEL-2 cell surface were found to be of various shapes and often showed variable profiles, e.g., extending small cytoplasmic processes along the surface of TEL-2 cells or appearing ameboidal. A remarkable feature of the TEL-2 cells was that they formed numerous "round spaces" at the surface of the TEL-2 cell layers. The thymocytes were often located around "round spaces," and some were seen migrating into TEL-2 cell layers through these round spaces. In addition, complementary examinations by transmission electron microscopy revealed that the internalization of thymocytes into TEL-2 cells occurs inside the TEL-2 cell layers after migration.(ABSTRACT TRUNCATED AT 250 WORDS)