Chemokine (C-C motif) receptor 5 (CCR5) is one of the major co-receptors for the macrophage (M)-tropic HIV-1. To prevent HIV-1 from entering into target cells, we inhibited CCR5 expression on target cell surface by recombinant adenovirus containing anti-sense CCR5 cDNA. A fragment of 653 bp cDNA located in the 5' region of CCR5 cDNA was reversely inserted into pAdTrack-CMV. Recombinant adenovirus containing antisense CCR5 cDNA (Ad-antiR5) was obtained by homologous recombination of resultant plasmid with the adenoviral backbone plasmid pAdEasy-2 in E. coli BJ5183 and then packed in AD-293 cells. Rate of positive CCR5 on U937 cell surface measured by flow cytometry was decreased from 89.53% to 1.88% after U937 cells infected with Ad-antiR5 for 24 hours, and this reduction lasted at least for 10 days. After challenged with HIV-1, the U937 cells infected with Ad-antiR5 produced much less p24 antigen in cultured medium than those infected with control recombinant adenovirus and the uninfected cells. The recombinant adenovirus had no effect on chemotactic activity and proliferation of the U937 cells. Therefore, the recombinant adenovirus containing anti-sense CCR5 cDNA can down-regulate CCR5 expression on U937 cells and protect the cells from HIV-1 infection without effects on their chemotaxis activity and proliferation function.