Construction and in vitro properties of chimeric simian and human immunodeficiency virus with the human TNF-alpha gene.
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.
Haga
Takeshi
T
Department of Veterinary Microbiology, Miyazaki University, Kibanadai Nishi, Miyazaki 889-2192, Japan. a0d518u@cc.miyazaki-u.ac.jp
Shimizu
Yuya
Y
Okoba
Masashi
M
Kumabe
Shino
S
Goto
Yoshitaka
Y
Shinjo
Toshiharu
T
Ichimura
Hiroshi
H
Kuwata
Takeo
T
Hayami
Masanori
M
Miura
Tomoyuki
T
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*
D017304:Annexin A5; D004274:DNA, Recombinant; D014409:Tumor Necrosis Factor-alpha