J.Jpn. Surg. Soc.. 93(4): 352-362, 1992

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Original article

A STUDY OF PERFORIN APPEARING IN HUMAN LYMPHOKINE-ACTIVATED KILLER (LAK) CELLS DERIVED FROM SPLEEN AND PERIPHERAL BLOOD MONONUCLEAR CELLS

Perforin is regarded as one of the main cytotoxic factors in such cells. We succeeded in cloning human perforin cDNA and recently used it to assess perforin appearing in LAK cells.
To derive the LAK cells, mononuclear cells separated from a human spleen were mixed with recombinant interleukin-2 and cultured. Almost no perforin mRNA appeared on day 0 but definitely accelerated on day 1 and tended to decrease on and after day 2. Peak of perforin mRNA observed on day 1 was found to precede cytotoxic activity on K562 and Daudi cells by one day. Similar results were found in LAK cells derived from peripheral blood mononuclear cells. Regarding the expression of perforin mRNA on day 0 as 1, the values on day 1 and 2 were calculated as 3.6 and 1.8, respectively. Immunological staining using an antiperforin antibody revealed perforin in the cytoplasm of large cells formed blasts, this also confirmed derivation of perforin on the protein level. Flow cytometry indicated that cells containing perforin included most CD16⁺ NK cells and some CD8⁺ T lymphocytes. The fact that TNF and IFN were simultaneously derived together with perforin suggested that the collective joint effect of these substances results in cytotoxic activity.

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