Technical aspects of lymphokine-activated killer cell production.
Adoptive immunotherapy is a novel approach to treating patients with cancer, utilizing as therapy a patient's own peripheral blood lymphocytes that have been activated by incubation with interleukin-2 (IL-2). These cells develop the ability to mediate tumor regression in vivo and are referred to as lymphokine-activated killer (LAK) cells. The production of LAK cells is a complex and labor-intensive process. Lymphocytes are collected by continuous-flow centrifugation, purified on Ficoll-Hypaque (FH) density gradients, incubated in vitro with IL-2, and then harvested for infusion into the patient. An automated approach to LAK cell generation has been developed using the Fenwal CS-3000 cell separator and polyolefin PL-732 blood storage bags. Lymphocyte concentrates (LC) containing 6.5 x 10(9) mononuclear cells per pack were obtained using standard leukapheresis techniques. Disposable apheresis kits were then modified to allow the LC to be pumped into the separation chamber along with a counter-centrifugal flow of saline, removing the platelets and plasma by elutriation. The remaining cells were underlaid with FH, displacing the lymphocytes into a collection bag, where they were washed and concentrated. Mean leukocyte recovery was 59.2% (99.9% lymphocytes, n = 14). The final product contained 6.7% of the initial platelets and had a hematocrit of less than 1%.(ABSTRACT TRUNCATED AT 250 WORDS)
Carter
C S
CS
Department of Transfusion Medicine, National Cancer Institute, Bethesda, MD 20892.
Leitman
S F
SF
Cullis
H
H
Muul
L M
LM
Nason-Burchenal
K
K
Rosenberg
S A
SA
Klein
H G
HG
D001331:Automation; D002469:Cell Separation; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007116:Immunization, Passive; D007694:Killer Cells, Natural; D007937:Leukapheresis / Q000295:instrumentation / Q000379:methods*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000187:drug effects; D008222:Lymphokines / Q000494:pharmacology*
D008222:Lymphokines