Modulation of the cytoskeleton and intracellular calcium in leukocytes exhibiting a cancer-associated chemotaxis defect.
Monocyte chemotaxis is severely depressed in patients with advanced tumors, but the cellular basis for this chemotactic defect is not known. Because the actomyosin cytoskeleton is thought to play a primary role in chemotaxis, we have employed flow cytometry to examine several aspects of the contractile machinery including myosin II, myosin light chain kinase (MLCK), actin, and cytoplasmic calcium in unstimulated and in formylpeptide-stimulated neutrophils and monocytes. Serum-pretreated polymorphonuclear leukocytes (PMNs) and monocytes from healthy blood donors or PMNs and monocytes isolated from tumor patients were studied. Leukocytes pretreated with serum from cancer patients exhibited decreased baseline myosin staining and a vastly different response to formylpeptide stimulation compared with leukocytes pretreated with normal human serum. In contrast, similar amounts of MLCK were observed in neutrophils and monocytes preincubated with normal or cancer serum with or without stimulation with formylpeptide. The fluorescent calcium indicator fluo-3 showed that resting and fMLP-stimulated levels of intracellular calcium were not significantly different in control and cancer serum-pretreated human leukocytes or in leukocytes isolated from tumor patients. Similarly, resting and fMLP-stimulated levels of F-actin in cancer patients' leukocytes as assessed by NBD-phallacidin staining did not differ significantly from those of normal leukocytes. Because the actomyosin cytoskeleton is intricately involved in leukocyte chemotaxis, alterations in the cytoskeleton may dramatically affect cell motility. The cytoskeletal alterations and changes in the response of leukocytes pretreated with cancer patients' serum to formylpeptide stimulation as described here may result in decreased chemotaxis by these cells.
Janeczek
A H
AH
Department of Anatomy and Cell Biology, University of Illinois at Chicago.
Van Alten
P J
PJ
Reyes
H M
HM
Walter
R J
RJ
D000199:Actins / Q000097:blood; D000328:Adult; D002118:Calcium / Q000097:blood*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D003599:Cytoskeleton / Q000648:ultrastructure*; D006258:Head and Neck Neoplasms / Q000097:blood*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D009000:Monocytes / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*; D009219:Myosin-Light-Chain Kinase / Q000097:blood; D009218:Myosins / Q000097:blood; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*; D012016:Reference Values
D000199:Actins; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009219:Myosin-Light-Chain Kinase; D009218:Myosins; D002118:Calcium