Objective: To determine the effect of β‐chemokines on HIV‐1 infection of primary macrophages, and to search for chemokine derivatives devoid of biological effects but efficient at protecting CD4+ T lymphocytes and macrophages against HIV‐1. Design: Use of chemically synthesized molecules devoid of biological contaminants and monocyte‐derived macrophages from healthy donors. Methods: Full‐length RANTES was chemically synthesized together with three derivatives, truncated of seven, eight and nine amino acids at the amino‐terminus ([8–68]RANTES, [9–68]RANTES and [10–68]RANTES), which were tested for their biological activity and antiviral effects. Results: Whereas full‐length and truncated RANTES derivatives bound to β‐chemokine receptor CCR‐5 with the same affinity as recombinant RANTES, the truncated forms were not chemotactic and acted as CCR‐5 antagonists in this respect, although a partial agonist effect was noted on cell metabolism. Full‐length RANTES and [8–68]RANTES protected T lymphocytes and macrophages from infection by HIV‐1, although 10‐fold higher concentrations of the truncated analogues were necessary to achieve the same effect as full‐length RANTES. With regard to the effect of RANTES on HIV‐1 infection of primary macrophages, our results contrast with most previously reported data. Conclusion: These data indicate that through binding to CCR‐5, truncated RANTES derivatives that are devoid of detectable biological effects may represent candidates as drugs to protect both lymphocytes and macrophages from HIV‐1.