Chemotaxis of T lymphocytes on extracellular matrix proteins. Analysis of the in vitro method to quantitate chemotaxis of human T cells.
The present report compares a variety of T cell purification protocols and chemotaxis procedures in assessing chemokine-induced T cell migration using a microchemotaxis assay. Rapidly purified T cells are capable of directly responding to the beta chemokines macrophage inflammatory protein-1 alpha (MIP-1 alpha), MIP-1 beta, and RANTES in the absence of alpha CD3 stimulation as previously described (Taub, D.D. and Oppenheim, J.J. (1993) Cytokine 5, 175). However, T cell purification schemes involving prolonged 37 degrees C incubations generally produce non-motile T lymphocytes that require stimulation with alpha CD3 antibody for 6-12 h in culture to recover chemotactic mobility. This loss of chemotactic potential appears to be due to prolonged 37 degrees C incubations as rapidly purified T cells lose migratory activity upon incubation at 37 degrees C. Radiolabeled binding analysis revealed that beta chemokine binding sites are downregulated as short as 2 h after incubation at 37 degrees C. T cells require the presence of extracellular matrix molecules to facilitate T cell migration. While many of these proteins permit chemotactic activity, human plasma and foreskin fibronectin were found to be the most effective matrix molecule for T cell migration. Kinetic analysis of T cell activation revealed that 6-12 h of anti-CD3 stimulation was optimal to restore the ability of purified T cells to migrate in response to the chemokines MIP-1 alpha, MIP-1 beta, RANTES, and IL-8. However, rapidly dividing T cells (> or = 48 h post alpha CD3 mAb stimulation) fail to migrate in response to any chemotactic stimulus. Together, these results suggest that the measurement of T cell migration, using microchemotaxis chambers, is a multifactorial process with strict environmental and activation requirements.
Taub
D D
DD
Clinical Support Laboratory, PRI/DynCorp, Inc., National Cancer, Institute-Frederick Cancer Research and Development Center, MD 21702-1201, USA.
Key
M L
ML
Clark
D
D
Turcovski-Corrales
S M
SM
D002448:Cell Adhesion; D002465:Cell Movement; D054407:Chemokine CCL4; D018946:Chemokine CCL5; D002633:Chemotaxis; D016207:Cytokines / Q000502:physiology; D005109:Extracellular Matrix / Q000502:physiology*; D006801:Humans; D016209:Interleukin-8 / Q000502:physiology; D008213:Lymphocyte Activation; D008222:Lymphokines / Q000502:physiology; D019402:Macrophage Inflammatory Proteins; D008722:Methods; D015846:Monokines / Q000502:physiology; D013601:T-Lymphocytes / Q000502:physiology*
D054407:Chemokine CCL4; D018946:Chemokine CCL5; D016207:Cytokines; D016209:Interleukin-8; D008222:Lymphokines; D019402:Macrophage Inflammatory Proteins; D015846:Monokines