Human lymphokine-activated killer (LAK) cells. I. Depletion of monocytes from peripheral blood mononuclear cells by L-phenylalanine methyl ester: an optimization of LAK cell generation at high cell density.
Pretreatment of peripheral blood mononuclear cells (PBMC) with 5 mM L-phenylalanine methyl ester (PheOMe) provides an efficient means to deplete monocytes. PheOMe does not affect the number of large granular lymphocytes after the pretreatment, but does inhibit natural killer cell cytotoxicity temporarily after the pretreatment. However, depletion of monocytes by PheOMe allows lymphokine-activated killer (LAK) cell generation with recombinant interleukin-2 (rIL-2) at high cell density (greater than 5 x 10(6) cells/ml). The time of the PheOMe pretreatment is 40-60 min, though some effect could be observed within 15 min, and the pretreatment could be performed at room temperature. Pretreatment density of PBMC with 5 mM PheOMe could be achieved at cell density up to 3 x 10(7) cells/ml. PheOMe-pretreated cells could be activated by rIL-2 in serumless media at high cell density. Pretreatment of PBMC with 5 mM PheOMe provides an efficient means to deplete monocytes, as compared to plastic and nylon-wool adherence. LAK cell generation is similar in both methods of monocyte depletion; therefore, depletion of monocytes allows, LAK cell generation at high cell density. The PheOMe procedure provides an improved and convenient process for preparing LAK cells for adoptive immunotherapy.
Leung
K H
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E.I. du Pont de Nemours and Co., Medical Products Department, Glenolden, PA 19036.
D002469:Cell Separation; D002478:Cells, Cultured; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D015979:Killer Cells, Lymphokine-Activated / Q000502:physiology*; D007700:Kinetics; D007958:Leukocyte Count; D008213:Lymphocyte Activation / Q000187:drug effects; D009000:Monocytes / Q000187:drug effects / Q000502:physiology*; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology
D007376:Interleukin-2; D011994:Recombinant Proteins; C034515:phenylalanine methyl ester; D010649:Phenylalanine