Abstract
A study was carried out to determine the sequence of events of T-cell mediated target cell lysis in microcinematography and electron microscopy. Highly efficient cytotoxic T lymphocytes (CTL) were generated in vivo and in vitro using preimmunized spleen cells and purification procedures. Such CTL were highly specific. This specificity correlated well with the number of adhesions formed between CTL and targets and this criterion was used to study killer-target cell interaction. Microcinematography showed that target cell lysis at the single cell level, despite time variations, could be clearly separated into three phases: (a) a recognition phase, visible by random crawling of CTL over the target cell surface until firm contact was established; (b) a post-recognition phase, during which firm contact between CTL and target was maintained without gross modification of either cell; (c) a phase of target cell disintegration, mainly characterized by vigorous blebbing of the cell membrane resulting in a motionless carcass of the target cell but not in its total dissolution. Only later this carcass decayed and formed a necrotic ghost. Electron microscopic observations were put into sequence according to microcinematography. Post-recognition phase was characterized by a tight apposition of the membranes of CTL and target cell. No gap junctions could be observed. During target cell disintegration, profound cytoplasmic and nuclear changes occurred simultaneous with surface blebbing. Most noticeable were extensive internal vacuolization, mitochondrial swelling, nuclear pycnosis and dissolution of the nucleolus. These observations suggested that target cell lysis does not start with a surface phenomenon similar to complement lysis, but a process involving practically the whole cell simultaneously. It is conceivable, therefore, that the signal from the CTL is transmitted across the target cell, and that the switch to sudden cell death is manipulated deep inside the cell.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adelstein E. H., Barrett B. A., Senhauser D. A. Ultrastructure of lymphocyte tumor cell interaction with localization of cell-bound antibody by ferritin labeling. Cancer Res. 1976 Feb;36(2 Pt 1):302–308. [PubMed] [Google Scholar]
- Brunner K. T., Mauel J., Cerottini J. C., Chapuis B. Quantitative assay of the lytic action of immune lymphoid cells on 51-Cr-labelled allogeneic target cells in vitro; inhibition by isoantibody and by drugs. Immunology. 1968 Feb;14(2):181–196. [PMC free article] [PubMed] [Google Scholar]
- Cerottini J. C., Brunner K. T. Cell-mediated cytotoxicity, allograft rejection, and tumor immunity. Adv Immunol. 1974;18:67–132. doi: 10.1016/s0065-2776(08)60308-9. [DOI] [PubMed] [Google Scholar]
- Ferluga J., Allison A. C. Observations on the mechanism by which T-lymphocytes exert cytotoxic effects. Nature. 1974 Aug 23;250(5468):673–675. doi: 10.1038/250673a0. [DOI] [PubMed] [Google Scholar]
- Henney C. S. Estimation of the size of a T-cell-induced lytic lesion. Nature. 1974 May 31;249(456):456–458. doi: 10.1038/249456a0. [DOI] [PubMed] [Google Scholar]
- Henney C. S. On the mechanism of T-cell mediated cytolysis. Transplant Rev. 1973;17(0):37–70. doi: 10.1111/j.1600-065x.1973.tb00123.x. [DOI] [PubMed] [Google Scholar]
- Julius M. H., Simpson E., Herzenberg L. A. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 1973 Oct;3(10):645–649. doi: 10.1002/eji.1830031011. [DOI] [PubMed] [Google Scholar]
- Kalina M., Berke G. Contact regions of cytotoxic T lymphocyte-target cell conjugates. Cell Immunol. 1976 Jul;25(1):41–51. doi: 10.1016/0008-8749(76)90095-2. [DOI] [PubMed] [Google Scholar]
- Lamelin J. P., Lisowska-Bernstein B., Matter A., Ryser J. E., Vassalli P. Mouse thymus-independent and thymus-derived lymphoid cells. I. Immunofluorescent and functional studies. J Exp Med. 1972 Nov 1;136(5):984–1007. doi: 10.1084/jem.136.5.984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liepins A., Faanes R. B., Lifter J., Choi Y. S., De Harven E. Ultrastructural changes during T-lymphocyte-mediated cytolysis. Cell Immunol. 1977 Jan;28(1):109–124. doi: 10.1016/s0008-8749(77)80011-7. [DOI] [PubMed] [Google Scholar]
- Matter A., Lisowska-Bernstein B., Ryser J. E., Lamelin J. P., Vassalli P. Mouse thymus-independent and thymus-derived lymphoid cells. II. Ultrastructural studies. J Exp Med. 1972 Nov 1;136(5):1008–1030. doi: 10.1084/jem.136.5.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matter A., Simpson E. The differentiation of cytotoxic T lymphocytes in vitro: an ultrastructural study. Cell Tissue Res. 1976 Feb 27;166(4):475–488. doi: 10.1007/BF00225912. [DOI] [PubMed] [Google Scholar]
- Matter A. The differentiation pathway of T lymphocytes. Evidence for two differentiated cell types. J Exp Med. 1974 Aug 1;140(2):566–577. doi: 10.1084/jem.140.2.566. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanderson C. J. The mechanism of T cell mediated cytotoxicity. II. Morphological studies of cell death by time-lapse microcinematography. Proc R Soc Lond B Biol Sci. 1976 Jan 20;192(1107):241–255. doi: 10.1098/rspb.1976.0011. [DOI] [PubMed] [Google Scholar]
- Seeman P. Ultrastructure of membrane lesions in immune lysis, osmotic lysis and drug-induced lysis. Fed Proc. 1974 Oct;33(10):2116–2124. [PubMed] [Google Scholar]
- Sellin D., Wallach D. F., Fischer H. Intercellular communication in cell-mediated cytotoxicity. Fluorescein transfer between H-2 d target cells and H-2 b lymphocytes in vitro. Eur J Immunol. 1971 Dec;1(6):453–458. doi: 10.1002/eji.1830010609. [DOI] [PubMed] [Google Scholar]