ABSTRACT
Chimeric simian and human immunodeficiency viruses (SHIVs) are useful tool for investigating AIDS pathogenesis and for development of vaccine. We constructed a SHIV-vpr vector (designated as SHIV-3sj) by replacing vpr region with restriction enzyme sites. SHIV-3sj was designed to express inserted gene along with its viral replication. Five cytokine genes were inserted into SHIV-3sj, and ability of viral replication and expression of the inserted genes were examined. The short insert including RANTES and IL-5 resulted in the successful expression from SHIV-3sj, while the construct having longer genes including IL-2, IL-6 and IL-12p35 failed to become replication competent. These results suggest that the length of the insert is an important factor for the replication ability of SHIV-3sj vector.
Subject(s)
DNA, Recombinant/genetics , Genes, vpr/genetics , Genetic Vectors/genetics , Genetic Vectors/physiology , HIV-1/genetics , Simian Immunodeficiency Virus/genetics , Virus Replication/genetics , Animals , Base Sequence , Cell Line , Chemokine CCL5/genetics , Gene Expression , HIV-1/physiology , Humans , Interleukin-12/genetics , Interleukin-2/genetics , Interleukin-5/genetics , Interleukin-6/genetics , Kinetics , Molecular Sequence Data , Open Reading Frames/genetics , Simian Immunodeficiency Virus/physiology , T-Lymphocytes/virologyABSTRACT
The positive effect of the co-expression of T helper (Th) cell type 2 cytokine interleukin-5 (IL-5) on nef-deleted simian/human immunodeficiency virus (SHIV) replication in vitro has been observed previously. To analyse whether the growth advantage of IL-5-containing SHIV (NI-IL5) in vitro would be relevant in vivo, the virus was inoculated into monkeys. Three rhesus macaques were inoculated intravenously with 10(4) TCID(50) of NI-IL5. Results were compared with those obtained previously from SHIV NM-3rN (intact) and SHIV-dn (nef-deleted)-infected monkeys. Cytokine production, analysed by IL-5 ELISA, showed a twofold increase in IL-5 concentration in the plasma soon after the peak of virus replication. Virus replication and antibody production were greater in monkeys inoculated with IL-5-expressing SHIV than in monkeys inoculated with nef-deleted SHIV without IL-5. These findings show a stimulation of SHIV replication by co-expression of IL-5 and suggest the important role of Th2-type cytokines in human immunodeficiency virus type 1 infection.
