Activation of blood T lymphocytes down-regulates CXCR4 expression and interferes with propagation of X4 HIV strains.
The chemokine receptor CXCR4 serves as a coreceptor for HIV-1 entry into CD4+ cells, in particular for strains emerging late in the infection. Cell surface expression of CXCR4 has, therefore, important implications for HIV-1 pathogenesis. Using blood lymphocytes cultured under various conditions, we studied the expression and regulation of CXCR4. Flow cytometry showed that only about 20% of freshly isolated lymphocytes expressed CXCR4 on the cell surface whereas in 80% of resting blood lymphocytes CXCR4 was located intracellularly. Within a few hours in culture, the intracellular CXCR4 was translocated to the surface and was expressed in the large majority of both naive and memory lymphocytes. A decrease in surface expression of CXCR4 was found when lymphocytes cultured overnight for maximal receptor expression were stimulated with phytohemagglutinin, anti-CD3 antibodies, phorbol 12-myristate 13-acetate and stromal cell-derived factor-1. The superantigen staphylococcal enterotoxin A, a more selective stimulus, induced a marked decrease in CXCR4 expression preferentially in cells positive for the CD25 activation marker. Confocal laser scanning microscopy demonstrated the presence of CXCR4 in the cytosol and on the surface of resting lymphocytes and also showed CXCR4 redistribution after activation. The number of cells infected by the X4 HIV strain NL4.3 paralleled the expression of CXCR4 in CD4+ T lymphocytes. Sustained reduction of CXCR4 cell surface expression upon activation with phytohemagglutinin correlated with a low number of CD4+ T lymphocytes expressing HIV p24 gag antigen. Our results indicate that activation of CD4+ T lymphocytes reduces surface expression of CXCR4 in part by receptor internalization and that cell activation-dependent CXCR4 down-regulation limits spread of infection by X4 viruses.
Bermejo
M
M
Servico de Microbiologia, Centro de Investigatión, Hospital Doce de Octubre, Madrid, Spain.
Martín-Serrano
J
J
Oberlin
E
E
Pedraza
M A
MA
Serrano
A
A
Santiago
B
B
Caruz
A
A
Loetscher
P
P
Baggiolini
M
M
Arenzana-Seisdedos
F
F
Alcami
J
J
D002478:Cells, Cultured; D015536:Down-Regulation; D015497:HIV-1 / Q000254:growth & development*; D006801:Humans; D007156:Immunologic Memory; D008213:Lymphocyte Activation; D008934:Mitogens / Q000494:pharmacology; D010835:Phytohemagglutinins / Q000494:pharmacology; D019718:Receptors, CXCR4 / Q000096:biosynthesis*; D013347:Subcellular Fractions; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism / Q000821:virology*
D008934:Mitogens; D010835:Phytohemagglutinins; D019718:Receptors, CXCR4