title: Construction and in vitro properties of chimeric simian and human immunodeficiency virus with the human TNF-alpha gene.
abstract: Tumor necrosis factor-alpha (TNF-alpha) has been reported to be involved in the development and progression of acquired immunodeficiency syndrome (AIDS). To study the role of this cytokine in AIDS pathogenesis, we constructed a chimeric simian and human immunodeficiency virus (SHIV) having the human TNF-alpha gene (SHIV-TNF) and characterized its properties in vitro. SHIV-TNF replicated both in M8166, a human T cell line, and in monkey peripheral blood mononuclear cells (PBMCs). Along with SHIV-TNF replication, TNF-alpha was detected in the culture supernatant by ELISA. The maximum expression level of TNF-alpha reached 120 ng/ml in M8166 cells, and 2.5 ng/ml in monkey PBMCs. The expressed TNF was biologically active, as shown by a cytotoxic assay using TNF-sensitive L929 mouse fibroblasts. This activity was detected at least until 10 passages of SHIV-TNF (74 days after the initial infection). In monkey PBMCs, SHIV-TNF replicated much better than the parental SHIV-NI. Flow cytometric analysis showed that the death of monkey PBMCs infected with SHIV-TNF was severer than that caused by the parental SHIV-NI. These results suggest that SHIV-TNF would be useful for inducing the disease in a monkey model, which may contribute to a better understanding of the role of TNF-alpha in AIDS etiology.
authors.affiliation: Department of Veterinary Microbiology, Miyazaki University, Kibanadai Nishi, Miyazaki 889-2192, Japan. a0d518u@cc.miyazaki-u.ac.jp
authors.forename: Takeshi
authors.initials: T
authors.lastname: Haga
authors.forename: Yuya
authors.initials: Y
authors.lastname: Shimizu
authors.forename: Masashi
authors.initials: M
authors.lastname: Okoba
authors.forename: Shino
authors.initials: S
authors.lastname: Kumabe
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Goto
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Shinjo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ichimura
authors.forename: Takeo
authors.initials: T
authors.lastname: Kuwata
authors.forename: Masanori
authors.initials: M
authors.lastname: Hayami
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Miura
mesh_terms: D000818:Animals; D017304:Annexin A5 / Q000032:analysis; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics; D006678:HIV / Q000235:genetics*; D006801:Humans; D008251:Macaca; D008957:Models, Genetic; D011995:Recombination, Genetic; D015302:Simian Immunodeficiency Virus / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000235:genetics*
chemical_list: D017304:Annexin A5; D004274:DNA, Recombinant; D014409:Tumor Necrosis Factor-alpha