Journal of Cell Science, Vol 89, Issue 4 561-573, Copyright © 1988 by Company of Biologists

JOURNAL ARTICLES

T-cell-mediated cytolysis: analysis of killer and target cell deformability and deformation during conjugate formation

C Foa, JL Mege, C Capo, AM Benoliel, JR Galindo and P Bongrand
Jeune Equipe CNRS, AIP 031 353, Marseille, France.

T-cell-mediated cytolysis is initiated by the formation of strong adhesions between killer and target cells. The present work was aimed at determining whether T lymphocytes might exert some mechanical stress on targets during the binding process. Target S194 myeloma cells were thus conjugated to cytotoxic T lymphocytes (CTLs) raised by mixed lymphocyte culture or a cloned lymphoid line that was no longer cytolytic (TG2OUA2). After incubation periods of various lengths, conjugates were processed for electron microscopy and micrographs were digitized for computerized analysis: the cell surface curvature (at the micrometre level) and rugosity (at the submicrometre level) were quantified in free and adhesion-involved regions. Also, the size of cell interaction areas and the distribution of intermembrane distances were measured. Finally, TG2OUA2 and target cells were aspirated into glass micropipettes with calibrated pressure in order to assay their resistance to deformation by mechanical forces. The following conclusions were suggested. (1) Formation of extensive contact zones (with a linear size of several micrometres) with tight intermembrane adhesion (more than 30% of the membrane contours in adhesive zones were separated by an apparent distance lower than 500 A) was essentially completed within less than one minute. (2) CTLs or TG2OUA2 cells were more villous than their targets, and they seemed to deform in adhesive zones in order to adapt to the target contour, rather than imposing some deformation on the target. This may be a general feature of actively adherent cells. (3) Some CTLs, but no TG2OUA2 cells, exhibited conspicuous protrusions extending towards the bound target. In this case, the target cell but not the CTL displayed markedly increased roughness in the adhesion area. (4) TG2OUA2 cells were more resistant to mechanical deformation than S194 target cells, in accordance with previous reports suggesting that the membrane of CTLs was more resistant than that of target tumour cells. Hence, CTLs might transiently impose mechanical stress on the target membrane during the course of lethal-hit delivery.

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