In vivo inactivation of Nef ITAM motif of chimeric simian/human immunodeficiency virus SHIVsbg-YE correlates with absence of increased virulence in Chinese rhesus macaques.

SHIVsbg, expressing Vpu, Tat, Rev, and Env proteins of HIV-1 Lai, was shown to be infectious for rhesus macaques. In this study, we mutated SHIVsbg Nef amino acids 17-18 from RQ to YE, conferring to SHIVsbg-YE the ability to replicate in vitro in unstimulated macaque PBMC. Juvenile macaques inoculated intravenously or orally with SHIVsbg-YE developed persistent infection. All macaques lost weight during the first 17 weeks but recovered afterward. All animals developed a strong HIV-specific humoral immune response. Viruses isolated 2 years postinoculation lost the ability to replicate in unstimulated macaque PBMC. Point mutations or 33-bp-wide deletions in the nef ITAM motif were responsible for this phenotype and correlated with clinical improvement of the infected macaques. These data demonstrate that the ITAM domain is inactivated in animals developing an acute antiviral immune response and may be detrimental to viral replication, perhaps by interfering with other well-conserved functions of SIV Nef protein.