Immobilized antibodies to CD45 induce rapid morphologic changes and increased tyrosine phosphorylation of p56lck-associated proteins in T cells.
CD45 is a transmembrane protein tyrosine phosphatase required for signal transduction through the Ag receptor complexes of T and B lymphocytes. Herein, we demonstrate that immobilized mAbs to the external domain of CD45 induce rapid and dramatic morphologic changes in a variety of T cell lines, including CD8+ cytotoxic clones. CD45-induced morphologic changes can be inhibited by the cytoskeletal inhibitors cytochalasin D and E and by the protein tyrosine kinase inhibitor herbimycin A. Consistent with the requirement for tyrosine kinase activity, tyrosine phosphorylation of proteins at about 60 to 75 kDa and 115 to 130 kDa is increased upon engagement with immobilized anti-CD45 mAb with kinetics paralleling the observed changes in morphology. The phosphorylation of these proteins is inhibited by tyrosine kinase inhibitors at concentrations that also inhibit changes in morphology. The phosphoproteins induced when cells are added to immobilized anti-CD45 are co-immunoprecipitated with p56lck, suggesting that this tyrosine kinase might play a role in the phosphorylation of these proteins. Consistent with this, there is no increase in the phosphorylation of these proteins in p56lck-deficient CTLL-2 cells in response to immobilized anti-CD45 mAb. An important role for p56lck in the morphologic pathway is further supported by the observation that p56lck-deficient human J.CAM1.6 cells, in contrast to the parental Jurkat line, cannot be induced to undergo morphologic changes. Taken together, these results suggest a possible role for CD45 in coordinating a cytoskeletal remodeling cascade that may be important in cell activation.
Arendt
C W
CW
Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada.
Hsi
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Ostergaard
H L
HL
D000199:Actins / Q000037:antagonists & inhibitors / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D016227:Benzoquinones; D017252:CD3 Complex / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000473:pathology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006825:Hybridomas; D047029:Lactams, Macrocyclic; D017493:Leukocyte Common Antigens / Q000276:immunology*; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051379:Mice; D010766:Phosphorylation; D011809:Quinones / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D019061:src-Family Kinases / Q000378:metabolism*
D000199:Actins; D000911:Antibodies, Monoclonal; D016227:Benzoquinones; D017252:CD3 Complex; D004791:Enzyme Inhibitors; D047029:Lactams, Macrocyclic; D011809:Quinones; D017828:Rifabutin; C020754:herbimycin; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D019061:src-Family Kinases; D017493:Leukocyte Common Antigens