We have previously shown the existence of primary human immunodeficiency virus type 2 isolates (MIC97 and MJC97) unable to use major coreceptors to entry into peripheral blood mononuclear cells, including CCR5 and CXCR4. We have now created a set of chimeric viruses derived from HIV-2(ROD), to study the contribution of env gene products in chemokine receptors usage and replication kinetics phenotype. The results obtained indicate that an env gene fragment, corresponding to the C1-C4 region of SU glycoprotein of both MIC97 and MJC97, impair efficient utilization of the major HIV coreceptors CCR5 and CXCR4 in phytohemagglutinin-stimulated peripheral blood mononuclear cells by ROD-MIC97 and ROD-MJC97 chimeric viruses. It also disrupts the ability to utilize established coreceptors for viral entry into GHOST-CD4 coreceptor-expressing cell lines. Resistance to CCR5 and CXCR4 inhibitors, as well as the ability to infect Delta32/Delta32ccr5 PBMC, was also observed in recombinant viruses containing the C1-C4 region from either MIC97 or MJC97. We also show that the presence of the TM region of env gene from MIC97 and MJC97 is sufficient to reduce viral replicative kinetics of ROD strain, indicating that this region, despite the presence and contribution of ROD genetic backbone, has an important role in viral progeny production efficiency.