Thrombospondin stimulates motility of human neutrophils.
Polymorphonuclear leukocytes (PMNs) migrate to sites of inflammation or injury in response to chemoattractants released at those sites. The presence of extracellular matrix (ECM) proteins at these sites may influence PMN accumulation at blood vessel walls and enhance their ability to move through tissue. Thrombospondin (TSP), a 450-kD ECM protein whose major proteolytic fragments are a COOH-terminal 140-kD fragment and an NH2-terminal heparin-binding domain (HBD), is secreted by platelets, endothelial cells, and smooth muscle cells. TSP binds specifically to PMN surface receptors and has been shown, in other cell types, to promote directed movement. TSP in solution at low concentrations (30-50 nM) "primed" PMNs for f-Met-Leu-Phe (fMLP)-mediated chemotaxis, increasing the response two- to fourfold. A monoclonal antibody against the HBD of TSP totally abolished this priming effect suggesting that the priming activity resides in the HBD of TSP. Purified HBD retains the priming activity of TSP thereby corroborating the antibody data. TSP alone, in solution at high concentrations (0.5-3.0 microM), stimulated chemotaxis of PMNs and required both the HBD and the 140-kD fragment of TSP. In contrast to TSP in solution, TSP bound to nitrocellulose filters in the range of 20-70 pmol stimulated random locomotion of PMNs. The number of PMNs migrating in response to bound TSP was approximately two orders of magnitude greater than the number of cells that exhibited chemotaxis in response to soluble TSP or fMLP. Monoclonal antibody C6.7, which recognizes an epitope near the carboxyl terminus of TSP, blocked migration stimulated by bound TSP, suggesting that the activity resides in this domain. Using proteolytic fragments, we demonstrated that bound 140-kD fragment, but not HBD, promoted migration of PMNs. Therefore, TSP released at injury sites, alone or in synergy with chemotactic peptides like fMLP, could play a role in directing PMN movement.
Mansfield
P J
PJ
Department of Pediatrics, Section of Hematology/Oncology, University of Michigan School of Medicine, Ann Arbor 48109.
Boxer
L A
LA
Suchard
S J
SJ
D002630:Chemotactic Factors; D002634:Chemotaxis, Leukocyte / Q000502:physiology; D015194:Diffusion Chambers, Culture; D006801:Humans; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009504:Neutrophils / Q000502:physiology*; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism / Q000502:physiology*; D019699:Thrombospondins
D002630:Chemotactic Factors; D010980:Platelet Membrane Glycoproteins; D019699:Thrombospondins; D009240:N-Formylmethionine Leucyl-Phenylalanine