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. 2013 Jul 29:4:218.
doi: 10.3389/fmicb.2013.00218. eCollection 2013.

Growth potentials of CCR5-tropic/CXCR4-tropic HIV-1mt clones in macaque cells

Naoya Doi  1 , Ayaka Okubo, Mizumo Yamane, Yosuke Sakai, Akio Adachi, Masako Nomaguchi
Affiliations

Growth potentials of CCR5-tropic/CXCR4-tropic HIV-1mt clones in macaque cells

Naoya Doi et al. Front Microbiol. .
No abstract available

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Figures

Figure 1
Figure 1
Characteristics of HIV-1mt clones. (A) Proviral genome structure of various HIV-1mt clones. Genomes of various HIV-1mt clones generated in our laboratory are schematically illustrated. White and black (a small portion of gag-capsid and an entire vif) areas stand for the genomic regions of HIV-1 NL4-3 (Adachi et al., 1986) and SIVmac MA239N (Shibata et al., ; Doi et al., 2010), respectively. The other colored regions are the sequences derived from various SIV/HIV-1s as shown. Restriction enzyme sites used for the insertion of these sequences into NL-DT5R are also shown. GenBank accession numbers are indicated in parentheses. Arrows represent the adaptive mutation sites in pol-integrase and env-gp120 that enhance virus growth potentials (Nomaguchi et al., 2013b). While NL-DT5R and MN4/LSDQgtu are CXCR4-tropic viruses, NL-DT562, NL-DT5AD, and MN5/LSDQgtu are CCR5-tropic. Although examined in macaque cells, the growth ability of NL-DT589 is not yet proven (see text). mac, rhesus macaques; gsn, greater spot-nosed monkeys. (B) Viral replication kinetics in M1.3S cells. Cell-free viruses were prepared from 293T cells transfected with proviral clones indicated, and equal amounts [5 × 106 and 5 × 105 reverse transcriptase (RT) units for left and right panels, respectively] were inoculated into M1.3S cells (2 × 106 cells). Viral replication was monitored at intervals by RT activity in the culture supernatants. The experiments were done as described previously (Kamada et al., 2006). M1.3S is the most refractory cell line to infection with SIVmac/HIV-1mt clones to the best of our knowledge, but is CD4-, CXCR4-, and CCR5-positive. NL-DT5R and NL-DT562 do not grow at all in M1.3S cells.
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