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Blood, 1 February 2002, Vol. 99, No. 3, pp. 978-984
IMMUNOBIOLOGY
Lipid rafts mediate biosynthetic transport to the T lymphocyte
uropod subdomain and are necessary for uropod integrity and
function
Jaime Millán,
María C. Montoya,
David Sancho,
Francisco Sánchez-Madrid, and
Miguel A. Alonso
From the Centro de Biología Molecular "Severo
Ochoa," Universidad Autónoma de Madrid, Consejo Superior de
Investigaciones Científicas and Servicio de
Inmunología, Hospital de la Princesa, Universidad
Autónoma de Madrid, Spain.
Polarized migrating T cells possess 2 poles, the uropod
protrusion at the rear and the leading edge at the front, with specific protein composition and function. The influenza virus hemagglutinin (HA) is a prototypical molecule that uses lipid rafts for biosynthetic transport to the apical surface in polarized epithelial Madin-Darby canine kidney (MDCK) cells. In this study, HA was used as a tool to
investigate the role of lipid rafts in vectorial protein traffic in
polarized T lymphocytes. Results show that newly synthesized HA becomes
selectively targeted to the uropod subdomain in polarized T
lymphoblasts. HA incorporates into rafts soon after biosynthesis, suggesting that delivery of HA to the uropod occurs through a pathway
of transport reminiscent of that used for its specific targeting to the
apical surface. HA and the adhesion molecules, intercellular adhesion
molecule 3 (ICAM-3), CD44, and CD43, 3 endogenous uropod markers, were
detected in surface rafts of T lymphoblasts. Cholesterol, a major
component of lipid rafts, was predominantly located in the uropod.
Disruption of lipid raft integrity by cholesterol sequestration
produced unclustering of ICAM-3 and the loss of uropodia and severely
impaired processes that require a polarized phenotype such as
intercellular aggregation and cell migration. Collectively, these
results indicate that lipid rafts constitute a route for selective
targeting of proteins to the uropod and that the rafts are essential
for the generation, maintenance, and functionality of T-cell
anteroposterior polarity.
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