Effect of lymphocyte conditioned medium on cytoplasmic membrane permeability and lysis of human tumor cells.

概要

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Cytolytic activity derived from conditioned medium (CM) of activated normal human peripheral blood lymphocytes was analyzed, using human solid tumor cells of kidney, bladder, breast and normal fibroblasts. Supplementation of CM to cell cultures resulted in an overall cytolysis, seen in cell number, cell growth, and colony formation. The lysis of cells was ascertained by the release of both lactic dehydrogenase and incorporated [3H] thymidine from the cells, to a degree which is directly proportional to the amount of cytolytic activity used and the length of exposure. The initial effect was revealed after one day and showed a greater than 85% cytoreduction by Day 3. The mechanism of cytolysis was further analyzed using human SK-RC-29 cells, which are suitable as target cells for the study of human cytolytic activity. CM was found to increase membrane permeability 6- to 8-fold by the flow cytometric determination of the permeability of a fluorescing DNA stain into the cells. The membrane permeability increase was seen prior to cell lysis, indicating its relationship to the destruction process. The cytolytic activity is non-dialyzable, heat stable at 60°C, and is distinct from human gamma interferon. Both T and B lymphocytes were shown capable of providing the cytolytic activity accompanied by increased membrane permeability.
The Keio Journal of Medicineの論文