Reorientation and fusion of cytotoxic T lymphocyte granules after interaction with target cells as determined by high resolution cinemicrography

The interaction of murine cytotoxic T lymphocyte (CTL) clones with human lymphoblastoid target cells was studied in thin preparations by using high resolution cinemicrography. CTL not bound to target cells were morphologically polar, possessing a broad leading edge containing the nucleus, and a tapered tail containing a large number of granules. The CTL were observed to move by the extension of pseudopods from the leading edge. Initial contact with a target cell was made via the leading edge of the CTL. If the human target cell expressed the appropriate HLA antigen, distinct morphologic changes occurred in the CTL as early as 2 min after initial contact. The CTL rounded up, and the nucleus moved from a position adjacent to the zone of contact to be replaced by the cytoplasmic granules. Redistribution of the granules was completed as early as 10 min after initial contact was made. These morphologic changes did not occur when the CTL made contact with other CTL, or with target cells that did not express the appropriate HLA antigens. In studies that make use of Nomarski optics, an apparent fusion of CTL cytoplasmic granules with the membrane in the vicinity of the target cell contact area was observed 4 min after binding, and before granule reorientation was complete. These data provide direct evidence for the occurrence of both reorientation of the cytoplasmic contents and granule fusion in CTL with a time course similar to that of administration of the lethal hit.