CCR5 and CXCR4 mediate Pyk2 phosphorylation in primary human macrophages treated with HIV-1 gp120.

(A) MDMs from a CCR5 wild-type donor were pretreated for 1 hour with (+) or without (−) the CXCR4 antagonist AMD3100 (1 μg/mL) before incubation for 5 minutes with R5 (JRFL) or X4 (IIIB) gp120 (2.5 μg/mL). (B) CCR5⁻ MDMs derived from donors homozygous for the CCR5 Δ32 deletion allele were treated for 5 minutes with JRFL or IIIB gp120 (2.5 μg/mL). Whole cell lysates were analyzed by immunoblotting with an antibody specific for the phosphorylated form of Pyk2 (upper panel, P-Pyk2) or with antibody specific for total Pyk2 (lower panel). Each experiment was repeated at least 3 times and the blots shown are representative of these experiments.