Cellular cholesterol enrichment impairs T cell activation and chemotaxis.
Human aging is associated with an increase in immune cell cholesterol levels, independent of circulating cholesterol levels. The effects of such an increase in membrane cholesterol on lipid raft-associated immune cell function have not been investigated. We sought to examine the effects of in vitro cholesterol loading on two known lipid raft-associated pathways of T cells, namely T cell activation and chemokine stimulation. Using beta-cyclodextrin (BCD) as a vehicle, we were able to rapidly load cholesterol onto human T cell lines and primary peripheral blood T cells without inducing significant cell toxicity. Loading of cholesterol to four-fold that of normal levels induced significant inhibition of intracellular calcium mobilization by both alphaCD3 and SDF-1alpha. Cholesterol-loaded peripheral T cells were completely unresponsive to alphaCD3/alphaCD28 stimulation, demonstrating no increase in IL-2, GM1 expression or cell size. T cell polarization of lipid rafts to alphaCD3/alphaCD28 beads was also impaired. In addition, cholesterol loading potently inhibited SDF-1alpha-induced chemotaxis. We propose that excess membrane cholesterol could potentially disrupt raft-related cell functions downstream of receptor triggering and that the loss of cholesterol regulation of aging immune cells could contribute to immune cell senescence.
Nguyen
Dzung H
DH
Laboratory of Immunology, National Institute on Aging, NIH, Intramural Research Program, 5600 Nathan Shock Drive, Baltimore, MD 21224-6825, USA. taubd@grc.nia.nih.gov
Espinoza
Juan C
JC
Taub
Dennis D
DD
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001692:Biological Transport / Q000502:physiology; D017252:CD3 Complex / Q000276:immunology / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D016764:Cell Polarity; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000502:physiology; D002634:Chemotaxis, Leukocyte; D002784:Cholesterol / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000187:drug effects / Q000502:physiology*; D021962:Membrane Microdomains / Q000502:physiology; D008863:Microspheres; D020033:Protein Isoforms / Q000276:immunology / Q000502:physiology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D020033:Protein Isoforms; D002784:Cholesterol; D002118:Calcium