Subject(s)
Carrier Proteins/toxicity , Carrier Proteins/therapeutic use , HSP70 Heat-Shock Proteins/toxicity , HSP70 Heat-Shock Proteins/therapeutic use , Neoplasms/drug therapy , Neoplasms/pathology , Animals , Carrier Proteins/administration & dosage , Cell Line, Tumor , Cell Proliferation/drug effects , HSP70 Heat-Shock Proteins/administration & dosage , Humans , Mice , Neoplasm Transplantation , Recombinant Proteins/administration & dosage , Recombinant Proteins/therapeutic use , Recombinant Proteins/toxicityABSTRACT
Human lymphokine-activated killer (LAK) cells were generated from peripheral blood lymphocytes (PBL) of normal volunteers by interleukin-2 (IL-2) stimulation for 1-8 days. During the first 3 days the surface marker CD16 characteristic for natural killer (NK) cells was expressed and later the CD3 marker characteristic for cytotoxic T cells became predominant. The conditioned media of LAK cells collected after interaction of LAK cells with K562 target cells was chromatographically separated into two cytotoxic fractions: F1 and F2. It was demonstrated that fraction F1 contained cytotoxic proteins having molecular weights of 30 and 40 kDa, and fraction F2 contained cytotoxic proteins having molecular weights of 22, 38 and 75 kDa. The presence of the proteins in each of these two fractions correlated with the phenotype changes of LAK cells: the F2 cytotoxic proteins were characteristic for NK-like cells, and the F1 proteins for cytotoxic T-lymphocyte (CTL)-like phenotypes.
Subject(s)
Killer Cells, Lymphokine-Activated/immunology , Proteins/metabolism , CD3 Complex/immunology , Cells, Cultured , Chromatography, Ion Exchange , Culture Media , Electrophoresis, Polyacrylamide Gel , Humans , Immunophenotyping , Interleukin-2/immunology , Killer Cells, Natural/immunology , Killer Factors, Yeast , Lymphocyte Activation/immunology , Molecular Weight , Receptors, IgG/immunology , T-Lymphocytes, Cytotoxic/immunology , Time Factors , Tumor Cells, CulturedABSTRACT
The influence of Ca2+ on the cytotoxic activity of natural killer cell cytotoxic factor (NKCF) was analyzed. When the natural killer susceptible cell line K562 was exposed to NKCF in the presence of 5 mM Ca2+, two peaks of cell damage were found. The first peak was observed after 30-40 min of incubation as a result of pore formation on the surface of target cells. The second was a peak of cytolytic activity which appeared after 24 h of incubation. Upon dilution of the NKCF preparation, only the first peak was observed. Therefore, NKCF produced by large granular lymphocytes in response to K562 consists of different proteins and represents pore-forming and cytolytic activities.
