Isolation and partial characterization of cytoplasmic granules of LAK cells from mice showing side effects of recombinant human IL-2.
The side effects of recombinant human interleukin-2 (rhIL-2) in mice are considered to be mediated by lymphokine-activated killer (LAK) cells that have cytoplasmic granules and infiltrate into target organs. In cloned cytotoxic lymphocytes (CLs) including LAK cells in vitro, granule exocytosis is one of the mechanisms in target cell lysis, and hemolytic and BLT-serine esterase (BLT-SE) activities detected in the granules are important to the cytotoxicity of the CLs in vitro. However, no information is available on these activities of LAK granules in vivo in mice showing the side effects of rhIL-2. Therefore, the present study was designed to isolate the LAK granules from the homogenate of LAK cell-rich subcutaneous tissue in mice treated with continuous infusion of a dose of rhIL-2 that induces known toxicity. High activities in hemolysis and BLT-SE were detected in Percoll fractions from the tissue homogenate. Electron microscopic observation of these fractions revealed almost only LAK granules. These results indicate that hemolytic and BLT-SE activities are detectable in the LAK granules in vivo, suggesting that the cells have potency for cytotoxicity via their granules and are involved in the toxicity of rhIL-2 in mice. In addition, perforin, which has been identified as a hemolytic protein in CL granules in vitro, was not detected by immunoblotting in the granule fraction of the LAK cells in vivo, indicating that the hemolytic activity of the LAK granules in vivo is not due to perforin.
Ueno
M
M
New Drug Research Laboratories, Shionogi & Co., Ltd., Futaba-cho 3-1-1, Osaka, Toyonaka, 561, Japan.
Yahara
I
I
Kishi
K
K
D000818:Animals; D002999:Clone Cells; D003594:Cytoplasmic Granules / Q000187:drug effects* / Q000648:ultrastructure; D003602:Cytotoxicity, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006461:Hemolysis / Q000187:drug effects; D015151:Immunoblotting; D007376:Interleukin-2 / Q000008:administration & dosage*; D015979:Killer Cells, Lymphokine-Activated / Q000187:drug effects*; D008562:Membrane Glycoproteins / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D011994:Recombinant Proteins / Q000008:administration & dosage; D013347:Subcellular Fractions / Q000187:drug effects / Q000648:ultrastructure
D007376:Interleukin-2; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D011994:Recombinant Proteins; D054353:Perforin