Construction and in vitro characterization of a chimeric simian and human immunodeficiency virus with the RANTES gene.
Chimeric simian-human immunodeficiency virus (SHIV) containing the env gene of HIV-1 infects macaque monkeys and provides basic information that is useful for the development of HIV-1 vaccines. Regulated-on-activation-normal-T-cell-expressed-and-secreted (RANTES), a CC-chemokine, enhances antigen-specific T helper type-1 responses against HIV-1. With the final goal of testing the adjuvant effects of RANTES in SHIV-macaque models, we constructed a SHIV having the RANTES gene (SHIV-RANTES) and characterized its properties in vitro. SHIV-RANTES replicated both in human and monkey T cell lines. Along with SHIV-RANTES replication, RANTES was detected in the supernatant of human and monkey cell cultures, at maximal levels of 98.5 and 4.1 ng/ml, respectively. A flow cytometric analysis showed that the expressed RANTES down-modulated CC-chemokine receptor 5 (CCR5) on PM1 cells, which was restored by adding anti-RANTES antibody. UV-irradiated culture supernatants from the SHIV-RANTES-infected cells suppressed replication of CCR5-tropic HIV-1 BaL in PM-1 cells. Differentiating real-time RT-PCR showed that pre-infection of SHIV-RANTES in C8166 cells expressing CCR5 suppressed the replication of HIV-1 BaL. Biological activity of the expressed RANTES and the inserted RANTES gene in SHIV-RANTES remained stable after 10 passages. These results suggest that SHIV-RANTES is worth testing in macaque models.
Shimizu
Yuya
Y
Department of Veterinary Microbiology, University of Miyazaki, 1-1 Kibanadai Nishi, Miyazaki 889-2192, Japan.
Okoba
Masashi
M
Yamazaki
Nanase
N
Goto
Yoshitaka
Y
Miura
Tomoyuki
T
Hayami
Masanori
M
Hoshino
Hiroo
H
Haga
Takeshi
T
D000818:Animals; D002460:Cell Line; D018946:Chemokine CCL5 / Q000235:genetics* / Q000378:metabolism*; D015536:Down-Regulation; D005818:Genetic Engineering; D006678:HIV / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D008252:Macaca fascicularis; D019713:Receptors, CCR5 / Q000378:metabolism; D011995:Recombination, Genetic; D015302:Simian Immunodeficiency Virus / Q000235:genetics* / Q000502:physiology*; D013997:Time Factors; D014779:Virus Replication
D018946:Chemokine CCL5; D019713:Receptors, CCR5